Average Optical Density Values Research Articles

Study on the effect of different durations of decalcification and depigmentation on programmed death ligand 1 immunohistochemical staining.

Programmed death ligand 1 (PD-L1) score is an important companion diagnosis to predict the response to immunotherapy. Immunohistochemistry can accurately assess the expression of PD-L1 in routine paraffin-embedded tissue. However, whether decalcified or depigmented tissue is still accurate and can be used as a companion diagnosis is controversial. This study attempts to resolve this controversy by analyzing the effects of decalcification and depigmentation at different times on PD-L1 expression. Placental tissues were selected for tissue microarray, decalcification was performed according to time gradients of 6, 12, 24, 36, and 48 h, and depigmentation was performed according to time gradients of 1, 5, 15, 30, and 60 min. The intensity of PD-L1 expression at different time points was observed and quantified. Ten PD-L1-positive esophageal squamous carcinoma samples were selected for decalcification treatment, and the PD-L1. Combined Positive Score (CPS), Tumor Proportion Score (TPS) and Immunocyte Proportion Score (IPS) and the positivity rates were compared before and after decalcification. After the placenta was decalcified, the intensity of PD-L1 positivity diminished, and the average optical density (AOD) value decreased with the prolongation of decalcification time and decreased significantly (P<0.05) at 24 h compared with the control group, and significantly (P<0.01) at 36 and 48 h compared with the control group. The intensity of PD-L1 positivity was weakened considerably after the treatment with potassium permanganate depigmentation. In addition, the AOD value decreased significantly (P<0.01) after the depigmentation time reached 5 min compared with the control group. Ten cases of PD-L1 positive esophageal squamous carcinoma were treated with 24 h decalcification, although the PD-L1 score decreased to a certain degree (P>0.05), and the positivity rate could reach 90%. After 36 h treatment, PD-L1 scores decreased, the CPS and IPS scores decreased significantly (P<0.05), and the positive rate was only 50%. Potassium permanganate depigmentation significantly reduces PD-L1 expression, even for a shorter time, affecting the accuracy of the results. The accuracy of PD-L1 remained high within 24 h decalcification. The above results have certain reference value for clinical selection of immunotherapy.

Ultraviolet C as a method of disinfecting medical silicone used in facial prostheses: An in vitro study – Part 2

Statement of problemDisinfection is an important factor in preserving facial prostheses and maintaining tissue health. However, whether disinfection with ultraviolet C is an effective disinfection method is unclear. PurposeThe purpose of this in vitro study was to evaluate the effectiveness of irradiation with different exposure durations of an ultraviolet-C light-emitting diode in the disinfection of the silicone (A-588–1; Factor II) used in facial prostheses. Material and methodsA total of 216 specimens were prepared, contaminated by multispecies biofilm, and divided into 9 groups (n=24) for different treatments: chlorhexidine 0.12% (G CHG), ultraviolet-C light-emitting diode for 5 minutes (G UVC5), ultraviolet-C light-emitting diode for 10 minutes (G UVC10), ultraviolet-C light-emitting diode for 20 minutes (G UVC20), their respective untreated controls (Gcontrol CHG, Gcontrol UVC5, Gcontrol UVC10, Gcontrol UVC20), and dimethyl sulfoxide (G DMSO) as the negative control. Cell viability was measured by using the methyl tetrazolium salt (MTT) method. Two statistical analyses were performed. First, a 2×3 ANOVA was carried out to compare the control groups (Gcontrol UVC5, Gcontrol UVC10, and Gcontrol UVC20) and the experimental groups of UV-C LED light with different exposure durations (G UVC5, G UVC10, and G UVC20). The second analysis was performed using generalized linear models to compare the optical density of the groups (G UVC5, G UVC10, G UVC20, G CHG, and G DMSO). ResultsCell viability results demonstrated a microbial reduction after exposure to the ultraviolet-C light-emitting diode for 20 minutes (G UVC20) compared with untreated controls (P<.05). The 5- and 10-minute exposures were statistically similar to their respective control groups (P>.05). The 20 minutes exposure had the lowest average optical density value, being statistically different from the 5-minute exposure (P<.05). A 20-minute exposure to the ultraviolet-C light-emitting diode (G UVC20) was similarly effective when compared with the standard disinfection treatment (G CHG) and dimethyl sulfoxide (G DMSO) (P>.05). ConclusionsIrradiation with an ultraviolet-C light-emitting diode for 20 minutes decreased the in vitro microbial cell viability on the medical silicone used in facial prostheses.

ELISA for leptospiral 3-hydroxyacyl-CoA dehydrogenase in urine is a promising screening tool for acute leptospirosis.

Introduction. Leptospirosis is a zoonotic disease that is prevalent worldwide. Leptospiral 3-hydroxyacyl-CoA dehydrogenase (3-HADH) is excreted in the urine of infected individuals. However, the potential use of 3-HADH as a biomarker for the diagnosis of leptospirosis using enzyme-linked immunosorbent assay (ELISA) has not been investigated. A technique that identifies Leptospira in a patient in urine sample will be valuable in regular diagnostics and epidemic scenarios, as opposed to existing serological approaches. This study aimed to develop and evaluate an ELISA that can detect 3-HADH in the urine of patients with confirmed acute leptospirosis and to assess its potential as a screening test for leptospirosis. Methods. Laboratory confirmation of acute leptospirosis was done by flaB-nested polymerase chain reaction (PCR) of plasma samples from suspected patients. ELISA-based determination of the presence of 3-HADH in the urine of PCR-positive patients versus PCR-negative patients matched for fever date was performed by coating ELISA plates with urine supernatants and using rabbit anti-3-HADH as the primary antibody. Receiver operating characteristic curve analysis was used to determine the cutoff values for the ELISA. The diagnostic measures between the PCR-positive and PCR-negative patients were compared using the Mann-Whitney U test. Results. In total, 158 febrile patients were assessed, of whom 121 (76.6 %) were male. Of the 15 flaB-nested PCR-positive patients, 12 were in the acute phase of the febrile illness. The best cutoff was an average optical density (ODav) value of 0.2200 for febrile patients. Sensitivity and specificity were 83.33% [95 % confidence interval (CI), 51.59-97.91 %) and 83.33 % (95 % CI, 76.05-89.13 %), respectively. The ODav values for PCR-positive patients in the acute phase of the disease (≤7 days of fever) were significantly higher than those for PCR-negative patients (P<0.001, U=114.0, z=-4.946). Conclusion. Detection of 3-HADH in urine by ELISA appears to be promising for the screening of acute leptospirosis in suspected patients.

Effect of electroacupuncture on enteric neuronal autophagy in functional constipation mice.

To explore the effect mechanism of electroacupuncture (EA) on functional constipation (FC) at the combined lower he-sea and front-mu points of large intestine based on enteric neuronal autophagy. A total of 40 SPF Kunming mice were randomly divided into 5 groups (n = 8), i.e. a control group, a model group, an acupuncture group, a 3-methyl adenine (3-MA) group, and a 3-MA + acupuncture group. Except the control group, the FC model was established by gavage with compound diphenoxylate suspension for 14 days in the other 4 groups. After successful modeling, the mice of the acupuncture group and the 3-MA + acupuncture group received EA at bilateral "Tianshu" (ST 25) and "Shangjuxu" (ST 37), stimulated for 30 min with disperse-dense wave, 2 Hz/15 Hz of frequency, 1 mA of intensity. EA was delivered once daily. One course of treatment was composed of 5 days and 2 courses were needed, with an interval of 2 days. An intraperitoneal injection of 3-MA (15 mg/kg) was administered 30 min before EA in the mice of the 3-MA group and the 3-MA + acupuncture group, once daily. Before and after intervention, the time of the first black stool defecation and defecation behaviors in 6 h were observed in each group. After intervention, in every group, the small intestine propulsion rate was calculated, the colon tissue morphology was observed using HE staining, the ultrastructure of enteric neuronal autophagy was observed under transmission electron microscope, and the expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1 and neuronal nuclear antigen protein (NeuN) in neurons of colonic muscularis were determined by immunohistochemistry. Before intervention, when compared with those in the control group, the time of the first black stool defecation was prolonged (P<0.01, P<0.05), and numbers (P<0.01), wet weight (P<0.01, P<0.05) and water content (P<0.05, P<0.01) of stool in 6 h were reduced in the model, acupuncture, 3-MA and 3-MA + acupuncture groups. After intervention, compared with those in the control group, the time of the first black stool defecation was longer (P<0.05), and numbers (P<0.01), wet weight (P<0.01) and water content (P<0.01) of stool in 6 h were decreased in the model group. The time of the first black stool defecation was shortened (P<0.01), and numbers (P<0.01), wet weight (P<0.01) and water content (P<0.01) of stool in 6 h were increased in the acupuncture group when compared with those in the model group. The time of the first black stool defecation was extended (P<0.01), and numbers (P<0.01), wet weight (P<0.01) and water content (P<0.01) of stool in 6 h were declined in the 3-MA + acupuncture group in comparison with those in the acupuncture group. All layers of colon tissue were normal and intact in each group. When compared with the control group, the small intestine propulsion rate and the average optical density (OD) values of LC3, Beclin-1 and NeuN in neurons of colonic muscularis were decreased (P<0.01), and autophagosomes were dropped in the model group. In the acupuncture group, the small intestine propulsion rate and the average OD values of NeuN, LC3 and Beclin-1 in neurons of colonic muscularis increased (P<0.01,P<0.05), and autophagosomes were elevated when compared with those in the model group. The small intestine propulsion rate and the average OD values of NeuN, LC3 and Beclin-1 in neurons of colonic muscularis were dropped (P<0.05,P<0.01) in the 3-MA + acupuncture group in comparison with those in the acupuncture group. Electroacupuncture may promote enteric neuronal autophagy and increase the number of neurons so that the intestinal motility can be improved and constipation symptoms can be relieved in FC mice.

Can the Acetabular Labrum Be Reconstructed With a Meniscal Allograft? An In Vivo Pig Model.

No single graft type has been shown to have a benefit in acetabular labral reconstruction. The native labrum and lateral meniscus share many similarities, suggesting that the meniscus may be a promising source of graft material in labral reconstruction. Using a pig model, we sought to evaluate the healing process of fresh-frozen meniscus allograft for acetabular reconstruction by assessing (1) MRI and macroscopic observations of the meniscus allograft; (2) histologic appearance and immunohistologic evaluation of the meniscus allograft, native meniscus, and labrum; (3) microscopic assessment of the native labrum and meniscus via scanning electron microscopy; and (4) biomechanical assessment of tensile properties. Twelve skeletally mature male miniature Bama pigs (24 hips) were randomly divided into two groups: labral defect group (control) and lateral meniscus allograft group. The selection of Bama pig specimens was based on the similarity of their acetabular labrum to that of the human acetabular labrum, characterized by the presence of fibrocartilage-like tissue lacking blood vessels. The pigs underwent bilateral hip surgery. Briefly, a 1.5-cm-long section was resected in the anterior dorsal labrum, which was left untreated or reconstructed using an allogeneic lateral meniscus. The pigs were euthanized at 12 and 24 weeks postoperatively, and then evaluated by macroscopic observations and MRI measurement to assess the extent of coverage of the labral defect. We also performed a histologic analysis and immunohistologic evaluation to assess the composition and structure of meniscus allograft, native labrum, and meniscus, as well as scanning electron microscopy assessment of the microstructure of the native labrum and meniscus and biomechanical assessment of tensile properties. Imaging measurement and macroscopic observations revealed that the resected area of the labrum was fully filled in the lateral meniscus allograft group, whereas in the control group, the labral defect remained at 24 weeks. The macroscopic scores of the meniscus allograft group (8.2 ± 0.8) were higher than those of the control groups (4.8 ± 1.0) (mean difference 3.3 [95% CI 1.6 to 5.0]; p < 0.001). Moreover, in the meniscus allograft group, histologic assessment identified fibrocartilage-like cell cluster formation at the interface between the graft and acetabulum; cells and fibers arranged perpendicularly to the acetabulum and tideline structure that were similar to those of native labrum could be observed at 24 weeks. Immunohistochemical results showed that the average optical density value of Type II collagen at the graft-acetabulum interface was increased in the meniscus allograft group at 24 weeks compared with at 12 weeks (0.259 ± 0.031 versus 0.228 ± 0.023, mean difference 0.032 [95% CI 0.003 to 0.061]; p = 0.013). Furthermore, the tensile modulus of the lateral meniscus allograft was near that of the native labrum at 24 weeks (54.7 ± 9.9 MPa versus 63.2 ± 11.3 MPa, mean difference -8.4 MPa [95% CI -38.3 to 21.4]; p = 0.212). In a pig model, lateral meniscus allografts fully filled labral defects in labral reconstruction. Regeneration of a fibrocartilage transition zone at the graft-acetabulum interface was observed at 24 weeks. The use of an autograft meniscus for labral reconstruction may be a viable option when labral tears are deemed irreparable. Before its clinical implementation, it is imperative to conduct a comparative study involving tendon grafts, which are extensively used in current clinical practice.

Contrast-Enhanced Ultrasound Combined With 2D Strain Imaging and Histopathological Multimodal Assessment of Carotid Plaque Vulnerability.

The aim of this study was to explore the value of contrast-enhanced ultrasound (CEUS) combined with 2-D strain imaging in evaluating carotid plaque vulnerability and the correlations among CEUS perfusion parameters, strain parameters and histopathological findings in different plaque segments. Patients with carotid artery stenosis who underwent carotid endarterectomy (CEA) at the First Affiliated Hospital of Xinjiang Medical University from September 2020 to June 2021 underwent preoperative carotid artery 2-D ultrasonography and CEUS. The plaques were divided into three segments: the proximal end of the shoulder, central cap and distal end of the shoulder. The peak intensity (PI) value and strain rate parameters of the regions of interest were analyzed. Plaques were divided into a stable group (8 cases) and an unstable group (19 cases). The microvascular density (MVD) and vascular endothelial growth factor (VEGF) expression of each patch in the unstable group were analyzed. The peak strain during the systolic period in each plaque segment in both groups showed the following pattern: proximal end shoulder > distal end shoulder > top (p < 0.05). The PI value for CEUS is also represented. In the unstable group, the PI values of each segment of the plaque were positively correlated with the MVD, near-center PI value and VEGF average optical density value. The average optical density of each segment was positively correlated with the MVD (p < 0.05). There were positive correlations between the PI values of the proximal and distal shoulder and the strain values (p < 0.05), and the MVD value of each segment, VEGF value and strain value were positively correlated (p < 0.05). PI and the pathological tissue components represented by CEUS were positively correlated with the mechanical parameters of the plaque along the long axis. There may be overlap between the high shear stress area of the plaque and the neovascular aggregation area, and the combination of the two has certain significance for assessing the vulnerability of the plaque.

Diagnostic challenges for Aelurostrongylus abstrusus infection in cats from endemic areas in Italy

BackgroundThe lungworm Aelurostrongylus abstrusus infects wild and domestic feline species worldwide and is considered a primary respiratory parasite of cats. Definitive diagnosis is based on the identification of first-stage larvae (L1s) released in faeces approximately 5 to 6 weeks after infection. More recently, serology has been shown to be a diagnostic alternative for A. abstrusus infection in cats. The present study aimed at evaluating the diagnostic performance of serological antibody detection compared to faecal examination for A. abstrusus infection in a population of cats with known infection status from endemic areas in Italy and to identify factors (larval scores, age, co-infections with other helminths) that may influence test sensitivity and specificity of serology.MethodsAll cats resulting positive using the Baermann technique (n = 78) were tested with the A. abstrusus ELISA. An additional 90 serum samples from cats living in three geographical areas with infection prevalence > 10%, but that resulted negative on Baermann, were also tested.ResultsAmong 78 cats copromicroscopically positive for L1s of A. abstrusus (Group 1), 29 (37.2%) were seropositive in ELISA. Of the 90 cats from Group 2 (cats living in three geographical areas in Italy with A. abstrusus prevalence > than 10%, but negative on Baermann examination), 11 (12.2%) were positive on ELISA. The overall seroprevalence was 23.8%. There was no statistical difference either between average optical density (OD) values of cats excreting > 100 L1s vs. cats excreting < 100 L1s (0.84 vs. 0.66; P value = 0.3247) or comparing the OD values with age of infected cats. Few Baermann-negative cats positive for Toxocara cati or hookworms were seropositive, supporting lack of cross-reactivity to these nematodes.ConclusionsResults from the present study suggest that relying solely on faecal examination may underestimate prevalence of A. abstrusus infection in cats and that field surveys based on antibody detection are useful for establishing true prevalence of infected and/or exposed animals.Graphical

The Expression of αvβ3 and Osteopontin in Osteoarthritic Knee Cartilage and Their Correlations With Disease Severity and Chondrocyte Senescence.

Osteoarthritis (OA) is the main joint disease associated with aging. Previous studies have confirmed that both osteopontin (OPN) and αvβ3 integrin are involved in the progression of knee OA. The purpose of this study was to determine the expression of OPN and αvβ3 integrin and chondrocyte senescence levels in OA. Forty-six cartilage tissues from normal and knee OA patients were divided into 4 groups of normal, minor, moderate, and severe lesions based on the Mankin score. Immunohistochemistry and western blotting were used to determine the expression of αvβ3, OPN, and senescent-associated-β-galactosidase (SAβ-gal) in articular cartilage. Then, Spearman's correlation was used to analyze the correlations between the Mankin scores and αvβ3, OPN and SAβ-gal. Pearson correlation analysis was used to analyze the correlations among αvβ3, OPN, and SAβ-gal. The expression of OPN, αvβ3, and SAβ-gal in articular cartilage was explored. αvβ3, OPN, and SAβ-gal proteins were all elevated in OA cartilage, and the correlation coefficient between the Mankin score and the average optical density value of αvβ3, OPN, SAβ-gal were r =0.60, r =0.75, and r =0.87, respectively, all P <0.001; the correlation between the average optical density value of αvβ3 and OPN was r =0.3191, P <0.05; the correlation between αvβ3 and SAβ-gal was r =0.4955, P <0.001; and the correlation between OPN and SAβ-gal was r =0.7821, P <0.001. The correlations among αvβ3, OPN, and SAβ-gal expression in articular cartilage might be important in OA progression and pathogenesis. Nonetheless, more research is needed to elucidate the exact contribution of αvβ3, OPN, and SAβ-gal to the degenerative process of OA.

THE EFFECTIVENESS OF PHALERIA MACROCARPA’S LEAF NANOEMULSION GEL ON STAPHYLOCOCCUS AUREUS BIOFILM THICKNESS (IN VITRO)

Background: Periodontal disease often occurs in the oral cavity with 75.6-78.3% at the age of 35-44 years. One of the causes of periodontal disease is the accumulation of Staphylococcus aureus biofilm in the early colonization of the formation of the dental pellicle. Phaleria macrocarpa, also known as Gods Crown, is a traditional plant with antibacterial properties that can be used in the health sector. Nanoemulsion gel technology has the advantage of increasing the stability of the material. Aim the study to determine the ratio of the effectiveness of Phaleria macrocarpa’s leaf nanoemulsion gel 10%, 20%, 30% to decrease Staphylococcus aureus biofilm thickness. Method: This research method was in vitro experimental laboratory research using a post-test control design. Thirty samples were divided into five groups: Phaleria macrocarpa’s leaf nanoemulsion gel 10%, 20%, 30%, positive control using chlorhexidine gluconate 0.2% and negative control using aqua dest. Samples were incubated for 4 and 8 hours. Optical density readings were carried out to see the biofilm thickness after being given a Gods Crown leaves nanoemulsion gel Results: The mean of Phaleria macrocarpa’s leaf nanoemulsion gel, 30%, produced the lowest average optical density value. The incubation time of 4 hours resulted in a lower optical density value than 8 hours. The Kruskal-Wallis was p&gt;0.05 showed there was no difference in biofilm thickness in each group. Conclusion: the effectiveness of the nanoemulsion group of Dewa crown leaf gel with a concentration of 30% was better than the 0.2% chlorhexidine gluconate gel and sterile distilled water group.

Effects of endoplasmic reticulum stress, liver function, insulin resistance and vascular endothelial function in patients with nonalcoholic fatty liver disease.

It is aimed to compare whether there are differences in endoplasmic reticulum stress, liver function, insulin resistance, and vascular endothelial function in patients with nonalcoholic fatty liver disease (NAFLD) with and without type 2 diabetes mellitus (T2DM). Forty patients with NAFLD, 38 patients with NAFLD combined with T2DM, and 30 patients with normal liver tissue were selected. They were set as Group A, Group B and Group C respectively. The expression level of glucose-regulated protein 94 (GRP94) and biochemical indicators such as alanine aminotransferase (ALT), free fatty acids (FFA), triglycerides (TG), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST), fasting blood glucose (FBG), and fasting insulin (FINS) were measured. Also, the calculation of the homeostatic model assessment for insulin resistance (HOMA-IR) index was performed. The reactive hyperemia index (RHI) was detected to evaluate the vascular endothelial function of patients. The comparison between groups and multi-factor analysis of the influencing factors of RHI was conducted. Compared with Group C, the expressions in Group A and Group B were distinctly enhanced (P <0.05). Also, the expression of GRP94 protein in Group B was distinctly higher than that in Group A (P <0.05). The average optical density values of Groups A, B, and C were 0.327 ± 0.007, 0.350 ± 0.009, and 0.299 ± 0.006, respectively. A comparison between the three groups was performed. The differences had statistical significance (P <0.05). The differences in the TG, ALT, AST, GGT, FINS and HOMA-IR between Group A and Group B had statistical significance (P <0.05). The RHI values of Groups A, B, and C were 1.59 ± 0.23, 1.79 ± 0.32, and 2.05 ± 0.47, respectively. A comparison between the three groups was performed. The differences had statistical significance (P <0.05). FFA, ALT and FBG in patients with NAFLD are risk factors for endothelial dysfunction (P <0.05). The liver damage caused by NAFLD may be related to the expression of GRP94. FFA, ALT and FBG are risk factors for endothelial dysfunction in NAFLD patients.

Age-Related Alterations of Hyaluronan and Collagen in Extracellular Matrix of the Muscle Spindles.

Background: Muscle spindles (MSs) play a crucial role in proprioception and locomotor coordination. Although the elasticity and viscosity of the extracellular matrix (ECM) within which MSs are embedded may play a key role in MS function, the impact of aging on ECM components is unclear. The aim of the current study was to investigate the age-related physiological changes of the ECM and to verify if these could be due to alterations of the environment directly surrounding MSs. Methods: Hematoxylin Eosin and picrosirius-red staining was carried out; collagen types I (COLI) and III (COLIII) were assessed, and biotinylated hyaluronan binding protein (HABP) immunohistochemical analysis was undertaken to evaluate alterations of the ECM in the intramuscular connective tissue (IMCT) of the hindlimbs of C57BL/6J male mice. Assessments were carried out on 6-week-old (Group A), 8-month-old (Group B), and 2-year-old (Group C) laboratory mice. Results: The capsule’s outer layer became progressively thicker with aging (it was 3.02 ± 0.26 μm in Group A, 3.64 ± 0.31 μm in Group B, and 5.81 ± 0.85 μm in Group C). The collagen in IMCT around and within the MSs was significantly higher in Group C, but there were no significant differences between Groups A and B. The MS capsules and continuous IMCT were primarily made up of COLI and COLIII. The average optical density (AOD) values of COLI in IMCT surrounding MS were significantly higher after aging (p < 0.05), but there were no significant differences in COLIII in the three groups (p > 0.05). HA was present in IMCT and filled the MSs capsule. The AOD of HABP of MS showed that there were lower HA levels in Group C with respect to Group A (p = 0.022); no significant differences were noted neither between Groups A and B nor between Groups B and C (p > 0.05). Conclusion: Age-related collagen accumulation and lower HA in the ECM in which the MSs were embedded may probably cause more stiffness in the ECM in vivo, which could help to partly explain the peripheral mechanisms underlying the age-related decline in functional changes related to MSs.

Effects of Electroacupuncture Preconditioning at Baihui and Zusanli Acupoints on the Expression of Inflammatory Factors in Brain and Intestines of Rats with Cerebral Ischemia/Reperfusion

<sec><title>Objective</title> To observe the effects of electroacupuncture preconditioning on the expression of interleukin-17A（IL-17A）and IL-10 in the brain and intestine of rats with cerebral ischemia/reperfusion and to explore the brain-gut protective effect of electroacupuncture preconditioning. </sec><sec><title>Methods</title> A total of 36 male Sprague-Dawley rats were randomly divided into the sham group (S-con group), the model group (M-con group), and the electroacupuncture preconditioning group (EP group) ,with 12 cases in each group. The S-con group was operated according to the model of middle cerebral artery occlusion/reperfusion (MCAO/R) with no blocking of the cerebral blood flow and reperfusion. The M-con group was given the operation of MCAO/R model and the cerebral blood perfusion was restored after 120 minutes. The EP group was intervened by electroacupuncture preconditioning at Baihui (DU20) and Zusanli (ST36) acupoints for two weeks, and was given the operation of MCAO/R model after the last electroacupuncture intervention for 24 hours, the cerebral blood perfusion was restored after 120 minutes. Before surgery for one day, after reperfusion for 12 h and after reperfusion for 1,3,7 days, the grip tester was used to measure the forelimb grasping strength; after reperfusion for 7 days, ELISA methods was used to determine the content of IL-17A and IL-10 in serum, Western blot method was used to detect the expression of IL-17A and IL-10 protein in ischemic cortex and colon tissue, immunohistochemistry method was used to calculate the average optical density (AOD) value of IL-17A and IL-10 in ischemic cortex and colon tissue, and HE staining method was used to observe the pathological change of ischemic cortex. </sec><sec><title>Results</title> ① Forelimb grasping strength: compared with the S-con group, the forelimb grasping strength of rats in the M-con group after reperfusion for 12 h, 1 d, 3 d, 7 d was decreased significantly (<italic>P</italic><0.05); compared with the M-con group, the forelimb grasping strength of rats in the EP group was increased significantly (<italic>P</italic><0.05). ② Content of IL-17A and IL-10 in serum: after reperfusion for 7 days, compared with the S-con group, the content of serum IL-17A and IL-10 of rats in the M-con group were significantly increased (<italic>P</italic><0.05); compared with the M-con group, the content of serum IL-17A in the EP group were significantly decreased (<italic>P</italic><0.05), and IL-10 in the EP group were significantly increased (<italic>P</italic><0.05). ③ Expression of IL-17A and IL-10 protein: after reperfusion for 7 days, compared with the S-con group, the expression of IL-17A and IL-10 protein in ischemic cortex and colon tissue in the M-con group were significantly increased (<italic>P</italic><0.05); compared with the M-con group, the expression of IL-17A in the EP group was significantly decreased (<italic>P</italic><0.05), and the expression of IL-10 protein in the EP group was significantly increased (<italic>P</italic><0.05). ④ AOD value of IL-17A and IL-10: after reperfusion for 7 days, compared with the S-con group, the AOD value of IL-17A and IL-10 in ischemic cortex and colon tissue in the M-con group were significantly increased (<italic>P</italic><0.05); compared with the M-con group, the AOD value of IL-17A in the EP group was significantly decreased (<italic>P</italic><0.05), the AOD value of IL-10 in the EP group was significantly increased (<italic>P</italic><0.05). ⑤ Pathological change of ischemic cortex: after reperfusion for 7 days, compared with the M-con group, the pathological change of ischemic cortex in EP group was significantly decreased (<italic>P</italic><0.05). </sec><sec><title>Conclusion</title> Electroacupuncture preconditioning at Baihui and Zusanli acupoints can reduce the pathological damage degree of the ischemic cortex and intestinal-central inflammation, improve neurobehavioral function, which may be related to brain-gut protection. </sec>

Consistency of pituitary adenomas: Amounts of collagen types I and III and the predictive value of T2WI MRI.

Pituitary adenomas, the most common type of lesion in the sellar region, rank third among all brain tumors, with an incidence of 73-94 cases per 100,000 individuals. Due to its high resolution, MRI is highly efficient in brain imaging and has emerged as the most appropriate method for tumor consistency evaluation. The present study aimed to assess the levels of collagen types I and III in pituitary adenomas with different consistencies and to determine the value of T2-weighted imaging (T2WI) MRI for predicting tumor consistency. A total of 55 patients with pituitary adenomas were divided into the soft and firm tumor groups according to intraoperative tumor consistency. The ratio of the tumor to Pons' signal intensities on T2WI scans was determined. A receiver operating characteristic curve was plotted to assess the specificity and sensitivity of T2WI in predicting tumor consistency. Average optical density (AOD) values for collagen types I (0.046±0.008 vs. 0.052±0.012, P=0.033) and III (0.044±0.008 vs. 0.050±0.010, P=0.016) were significantly lower in the soft tumor group compared with those in the firm tumor group. There was no significant difference in the ratio of the tumor to Pons' signal intensities on T2WI scans. The area under the ROC curve was 0.595±0.078 (P=0.250). The maximum tumor diameter significantly differed between the soft and firm tumor groups (P=0.001). AOD values for collagen types I and III were significantly correlated with the maximum tumor diameter (P<0.001). The amounts of collagen types I and III were elevated in firm pituitary tumors compared with the soft ones. The ratio of tumor to Pons' signal intensities on T2WI scans was not able to accurately predict tumor consistency. The size of pituitary adenomas may be associated with the expression levels of collagen types I and III.

Efficacy Analysis of Bronchoscopic Interventional Therapy for Benign Tracheal Stenosis by Combination of Intelligent Imaging Analysis System and Immunohistochemistry

Objective: To assess the efficacy of bronchoscopic interventional therapy for benign tracheal stenosis by combination of intelligent imaging analysis system and immunohistochemistry. Methods: The therapeutic effect was evaluated by Computed Tomography (CT) image changes, and the effects of thermofrequency and glucocorticoids were analyzed by animal experiments. Eighteen patients with benign tracheal stenosis who were enrolled in 2017.06–2019.06 were enrolled, and the thickness, diameter and lumen area of the most narrow tracheal tube before and after bronchoscopy and 1 month after bronchoscopy were measured using intelligent image analysis software. Thirty male SD rats were randomly divided into three groups (Radiofrequency group, Glucocorticoids+Radiofrequency group and control group). At the 5th, and 28th day the comparisons were made on the pathological changes of rats skin,the average optical density value of TGF-β1 and the collagen density after the damage by Radiofrequency, Glucocorticoids+Radiofrequency. Results: The thickness, diameter and lumen area of the wall before and after bronchoscopy are: 3.17 ± 1.01 mm versus 1.60 ± 0.23 mm; 3.61 ± 1.23 mm versus 9.04 ± 1.00 mm; 11.19 ± 5.32 versus 67.34 ± 13.80 mm2. The scar healing was observed in Radiofrequency group, but in the Glucocorticoids+Radiofrequency group the rats’ skin were healed incompletely. The results showed significant statistical differences among three groups on TGF-β1 comparisons: 988.41 ± 111.91, 730.19 ± 109.52 and 437.37 ± 60.48 (P &lt; 0.05). The same results of comparisons on the collagen density showed: 96540.58 ± 29909.51, 43768.16 ± 6264.72 and 34521.15 ± 4636.19 (P &lt; 0.05). Conclusions: After bronchoscopy intervention, the wall thickness of benign tracheal stenosis can be reduced, and the lumen diameter and area can be increased. The results can be analyzed using CT. Locally injected Betamethasone can alleviate the cicatricial tissue hyperblastosis induced by radiofrequency.

ОПТИМИЗАЦИЯ ПРОТОКОЛА ИММУНОФЕРМЕНТНОГО АНАЛИЗА ДЛЯ ИНДИКАЦИИ Т-2 ТОКСИНА

T-2 toxin is one of the most common and toxic trichothecene mycotoxins – secondary metabolites of molds that develop on cereals and some other crops. This article discusses the development stage of a test system for the determination of T-2 toxin based on enzyme-linked immunosorbent assay (ELISA), which uses an enzyme label – a conjugate of anti-rabbit goat antibodies with peroxidase. An important step in the creation of any ELISA-based test system is the preliminary titration of reaction components. The aim of the work was to determine the optimal concentration of the conjugate of antispecies antibodies in an indirectly competitive enzyme-linked immunosorbent enzyme immunoassay with the indication of T-2 toxin. A number of dilutions of the antivirus conjugate were examined: 1 : 1000; 1 : 2000; 1 : 3000; 1 : 4000; 1 : 5000; 1 : 7500; 1 : 10000; 1 : 12 500; 1 : 15,000; 1 : 17,500; 1 : 20,000; 1 : 30 000. In the ELISA staging protocol, calibration solutions of T-2 toxin at concentrations of 0.0 were used; 2.5; 5, 10, 20, 40, 80, 160 ng/ml and specific polyclonal rabbit antibodies to the T-2-BSA conjugate. Based on the average optical density values, calibration plots were constructed using the percentage of signal absorption from the zero standard. When evaluating the results of the study, the criterion for choosing the dilution of the conjugate of anti-species antibodies was considered the greatest, at which the best linearity of the grading plot is achieved and the level of its non-specific reaction with the zero standard would be the lowest. It was established that the optimal variants of dilutions of the conjugate of anti-species antibodies under the same experimental conditions tested were 1: 4000, 1 : 5000 and 1 : 12 500. Dose-dependent signal absorption was observed in all concentrations of anti-species antibodies. Dilutions of the conjugate of anti-species antibodies 1 : 1000–1 : 3000 and 1 : 17 500–1 : 30 000 were not taken into account, since the optical density of most wells was higher than the optimal boundaries in the first case (&gt; 3.9) and lower in the second (&lt; 0.4). Based on the foregoing, optimal dilution of the conjugate of anti-species antibodies was selected 1 : 12 500.

Simulation analysis and preparation of a high optical density laser protection filter.

High optical density (OD) filters have been widely used in space observation, optical detection, and laser protection. However, the lack of high OD value filters is restricting their application. This paper reports the formulation of a three-dimensional mesh model that can help predict the effects of pinhole defects, thickness error, and uniformity on the transmittance and OD value of optical filters. A laser protection filter (LPF) with a high OD value was prepared on fused silica using a microwave plasma-assisted pulsed DC reactive sputtering technique. The transmittance and OD value of the LPF were measured. Comparing the designed, measured, and simulated results, we found that the thickness error and uniformity of the layers mainly affected the passband transmittance of the LPF and had little effect on the OD value of the blocking band. In contrast, the pinhole defects were the main factor that decreased the OD value of the blocking band. The average OD values of the prepared LPF in the blocking bands of 527-532 and 755-833 nm were 8.832 and 10.191, respectively. By comparing the transmittance and OD value of the simulated and measured results, we found that the LPF has ${\pm 1}\% $±1% uniformity error and 0.5% pinhole ratio. Suggestions for preparing high OD optical filters are provided, and further improvements are summarized.

Neural Remodeling of the Left Atrium in Rats by Rosuvastatin Following Acute Myocardial Infarction.

ObjectiveThis study aims to investigate the effect of rosuvastatin on sympathetic neural remodeling of the left atrium (LA) in rats after myocardial infarction (MI).MethodsRats were randomly divided into a three groups: sham group, statin group, and MI group. The mRNA expression levels of the growth-associated protein-43 (GAP43) and nerve growth factor (NGF) were measured by RT-PCR. Immunohistochemistry was used to detect the distribution and density of GAP43- and NGF-positive nerves. The expression levels of these proteins were quantified by Western blot.ResultsCompared with the sham group, the average optical density (AOD) values of GAP43 and nerve growth factor (NGF)-positive substances in the LA in the statin and MI groups were significantly higher (P<0.01), but the AOD values in the statin group were lower than of those in the MI group (P<0.01). Furthermore, the AOD values of GAP43 and NGF positive nerves in the left stellate ganglion in the statin and MI groups were significantly higher (P<0.01), but the AOD values in the statin group were lower, when compared with the MI group (P<0.01).ConclusionRosuvastatin could effectively improve the sympathetic neural remodeling of LA in MI rats.

LncRNA HULC promots HCC growth by downregulating miR-29

Objective: To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down-regulating miR-29. Methods: The expression levels of HULC and miR-29 in HCC tissues and cells were detected by real-time quantitative PCR (RT-qPCR), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR-29 and its target gene SETDB1 were determinate by RT-qPCR. According to the bioinformatic prediction of the miR-29 binding site in the HULC sequence, the report gene plasmids were constructed. The HCC cells were co-transfected with miR-29 mimics or miR-29 inhibitor, and the HULC targeted regulation of miR-29 was verified by dual luciferase reporter assay. The effect of miR-29 on the HULC-mediated proliferation in HCC cells was detected by cell count kit 8 (CCK-8) experiment. Expression of tumor proliferation antigen Ki-67 was detected by RT-qPCR.The Hep3B cells were inoculated in mice and miR-29 mimics and miR-29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki-67 in tumor tissues were detected by immunohistochemical staining. Results: The expression of HULC was significantly up-regulated while the expression of miR-29 was significantly down-regulated in HCC tissues and cells (P<0.01). The level of HULC was negatively correlated with miR-29 in tumor tissues (r=-0.754, P<0.01) and HCC cells (r=-0.865, P<0.05). The in vitro experiments showed that, compared with the blank control group, the expression of miR-29 in HULC overexpressed Huh7 cells was significantly reduced, while the mRNA level of miR-29 target gene SETDB1 was increased (P<0.01). The expression of miR-29 was significantly increased in HULC deleted Hep3B cells, while the mRNA expression of SETDB1 was decreased (P<0.01). Double luciferase reporter gene assay showed that miR-29 mimics significantly inhibited the luciferase activity of Hep3B cells transfected with HULC wide type (psi-HULC-WT) plasmid but had no effect on Hep3B cells transfected with mutant plasmid (psi-HULC-Mut). However, the miR-29 inhibitor antagonized the inhibitory effect of miR-29 mimics on luciferase activity of psi-HULC-WT (P<0.01). Cell proliferation experiments showed that, compared with the control group, the proliferation ability of miR-29 mimics overexpressed Huh7 cells was significantly reduced.After 24, 48 and 72 hours of treatment, the proliferation rates of Huh7 cells in the HULC overexpressed group were (43.87±3.82)%, (83.45±7.46)% and (123.34±8.67)%, respectively, significantly higher than (13.45±1.77)%, (23.54±1.37)% and (38.21±2.09)% of control group (P<0.05). After treatment for 48 and 72 hours, the proliferation rates of miR-29 mimics transfected Huh7 cells were (57.10±1.94)% and (73.76±3.46)%, respectively, significantly lower than (83.45±7.46)% and (123.34±8.67)% of control group (P<0.05). After treatment for 48 and 72 hours, the proliferation rates of Huh7 cells transfected with miR-29 mimics and miR-29 inhibitor group were (76.45±3.24)% and (89.37±4.37)%, respectively, significant higher than (57.10%±1.94)% and (73.76±3.46)% of the control group (P<0.05). After 48 h transfection, the expression of Ki-67 in Huh7 transfected with miR-29 mimics was significantly inhibited compared with the control group (P<0.01). However, the expression of Ki-67 mRNA was increased in Huh7 cells transfected with miR-29 inhibitor (P<0.01). The results of in vivo experiments showed that the tumor volumes of the control group, miR-29 mimics group and miR-29 mimics + miR-29 inhibitors group were (504.0±19.6) mm(3), (310.0±24.3) mm(3) and (483.7±21.2) mm(3), respectively. Injection of miR-29 mimics reduced while miR-29 inhibitor promoted tumorigenesis ability of Huh7 in nude mice (P<0.01). The immunohistochemical staining showed that the average optical density values of Ki-67 protein in tumor tissues of the control group, miR-29 mimics group and miR-29 analogue+ miR-29 inhibitor group were 0.65±0.08, 0.36±0.07 and 0.56±0.06, respectively. The expression level of Ki-67 protein in miR-29 mimics group was significantly reduced (P<0.01) while increased in the miR-29 mimics+ miR-29 inhibitor group (P<0.01). Conclusion: LncRNA HULC promotes HCC growth by down-regulating miR-29.

The Effect of Buddleja officinalis Maxim Eye Drops on Morphology and Apoptosis in Lacrimal Gland of Experimental Dry Eye Rabbit Model.

The purpose of this study was to investigate the effects of Buddleja officinalis Maxim eye drops on morphology and apoptosis in lacrimal glands of the experimental dry eye rabbit model. A total of thirty-six male rabbits were divided into six study groups, consisting of the control group and the dry eye rabbit model group (without any treatment), the dry eye rabbit model group treated with testosterone, and the dry eye rabbit model group treated with different concentrations of Buddleja officinalis Maxim eye drops (1.0 mg/ml, 1.5 mg/ml and 3.0 mg/ml). The lacrimal glands were evaluated by hematoxylin-eosin staining and immunohistochemistry. Buddleja officinalis Maxim eye drops can improve the morphological structure of the lacrimal gland in the dry eye model of castrated rabbits. The average optical density values of PI3K, Akt, and caspase-9 protein in the lacrimal gland tissue of the 3 mg/ml Buddleja officinalis Maxim eye drops group were significantly different from those in the model group (P < 0.01) and similar to the testosterone control group and the control group (P > 0.05). Buddleja officinalis Maxim eye drops can improve the morphological structure of the lacrimal gland in the dry eye model of castrated rabbits.

Mimic Cochlear Implant Surgery-Induced Cochlear Infection Fails to Further Damage Auditory Pathway in Deafened Guinea Pigs.

BackgroundKanamycin and subsequent furosemide administration was applied to the healthy guinea pigs to induce deafness.Material/MethodsOf the deafened guinea pigs, 10 were further infused with anti-infection procedures (Group B) and the other 10 animals did not undergo anti-infection procedures (Group C). In Group B, the deafened animals were able to restore cochlear and middle ear functions following the anti-infection procedure. In Group C, all animals developed cochlear and middle ear infections.ResultsCompared to the healthy guinea pigs, hair cells and spiral ganglion neurons (SGN) of deafened animals (in Group B and Group C) were severely damaged. SGN density of deafened animals was significantly lower than that of healthy control animals in all ear turns except the basal turn. There was no significant difference between Group B and Group C in SGN density. The average optical density value of neurofilaments of deafened animals was also significantly decreased after the ototoxic drug administration. Notably, the density of the neurons in the cochlear nucleus region (CNR) of the brainstem were not significantly different between the healthy control guinea pigs and deafened animals.ConclusionsMimic cochlear implant surgery-induced cochlear infection caused no significant damage to the auditory pathway in ototoxic drug-induced deafened guinea pigs.