Obvious Inflammatory Reaction Research Articles

A silk-based hydrogel containing dexamethasone and lipoic acid microcrystals for local delivery to the inner ear

Local drug delivery has been exploited recently to treat hearing loss, as this method can both bypass the blood-labyrinth barrier and provide sustained drug release. Combined drug microcrystals (MCs) offer additional advantages for sensorineural hearing loss treatment via intratympanic (IT) injection due to their shape effect and combination strategy. In this study, to endow viscous effects of hydrogels, nonspherical dexamethasone (DEX) and lipoic acid (LA) MCs were incorporated into silk fibroin (SF) hydrogels, which were subsequently administered to the tympanic cavity to investigate their pharmaceutical properties. First, we prepared DEX and LA MCs by a traditional precipitation technique followed by SF hydrogel incorporation (SF+DEX+LA). After characterization of the physicochemical features, including morphology, rheology, and dissolution, both a suspension of combined DEX and LA MCs (DEX+LA) and SF+DEX+LA were administered to guinea pigs by IT injection, after which the pharmacokinetics, biodegradation and biocompatibility were evaluated. To our surprise, compared to the DEX+LA group, the pharmacokinetics of the SF+DEX+LA hydrogel group did not improve significantly, which may be ascribed to their nonspherical shape and deposition effects of the drugs MCs. The cochlear tissue in each group displayed good morphology, with no obvious inflammatory reactions. This combined MC suspension has the clear advantages of no vehicle, easy scale-up preparation, and good biocompatibility and outcomes, which paves the way for practical treatment of hearing loss via local drug delivery.

Bionic artificial penile Tunica albuginea

Bionic artificial penile Tunica albuginea

Preparation of Cross-Linked Bovine Tendon Acellular Fibers and Study of Their Biophysical and Chemical Properties

To explore the effect of glutaraldehyde (GA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) cross-linking on the biophysical and chemical properties of acellular scaffold to better provide suitable donor materials for tendon reconstruction. GA and EDC with different concentrations and action time gradients were used to cross-link the acellular scaffold. By detecting the collagen content in the cross-linked scaffold and the cytotoxicity, the cross-linking scheme with minimal damage to the scaffold and minimal cytotoxicity was explored for subsequent studies. The biomechanical properties (durability, elastic modulus, stressmax) of the scaffolds in GA, EDC, acellular scaffold, and tendon groups were compared, and the scaffold rat models were constructed to further evaluate their in vivo histocompatibility. Under different concentration gradients, the collagen content of the scaffolds in the GA and EDC groups had no obvious difference. When 0.5% GA was cross-linked for 24 h, and the mass ratio of EDC (1:2) was cross-linked for 4 h, the inhibition rate of the scaffold extract on fibroblasts was the lowest. In the mechanical property test, the Stressmax, durability, and elastic modulus of the cross-linked acellular scaffolds were significantly improved than those before cross-linking, and the elastic modulus of the EDC acellular scaffold was similar to that of the bovine tendon. In the compatibility test, compared with the acellular scaffold group, fibroblast activity in the GA group decreased obviously, and the scaffold implanted in rat models led to a persistent chronic inflammatory reaction. However, cells in the EDC group could maintain good activity. Moreover, the scaffold had good compatibility with rats and did not cause an obvious inflammatory reaction. EDC cross-linking scheme will not damage the acellular scaffold, and the cytotoxicity of the obtained scaffold is controllable. Additionally, EDC cross-linked acellular scaffold has mechanical properties similar to normal tendons and excellent histocompatibility.

In Vitro and In Vivo Analysis of the Effects of 3D-Printed Porous Titanium Alloy Scaffold Structure on Osteogenic Activity.

The effect of titanium scaffold geometry on the bone regeneration ability of the scaffold remains unclear. Here, selective laser melting as a 3D printing technology was used to create porous titanium alloy scaffolds with two unit structures: a hollow hexagonal prism (group A) and a hollow triangular prism (group B). The structures and morphologies of the scaffolds were characterized before mechanical properties were simulated. Cell adhesion behaviors, osteoblast activity and proliferation, and alkaline phosphatase (ALP) activity were evaluated, in addition to in vivo testing in an animal model. The results showed that the two scaffolds made of Ti6Al4V had compression moduli similar to that of human cortical bone (116.91 ± 0.01 and 174.29 ± 2.21 MPa vs. 89–164 MPa). The two scaffolds were nontoxic to cells and had good biocompatibility, while group A scaffolds facilitated cell adhesion. The number of cells increased gradually in culture. The ALP activity of cells on group A scaffolds demonstrated higher osteogenic ability than that of group B scaffolds. The in vivo tests showed that all scaffolds retained their shape well after implantation, and no obvious inflammatory reaction or infection in surrounding tissues was found. Based on fluorescence staining, mature new bone formation was found at week 12. Group A scaffolds showed better bone integration ability compared with group B scaffolds. The percentage of new bone area in group A (7.5%) was higher than that in group B (6.5%). This research suggests that the hollow hexagonal prism structure of porous scaffolds can promote osteogenic differentiation and osseointegration better than the triangular prism structure.

The influence of yttrium and manganese additions on the degradation and biocompatibility of magnesium-zinc-based alloys: In vitro and in vivo studies

The repair and regeneration of bone defects are highly challenging orthopedic problems. Recently, Mg-based implants have gained popularity due to their unique biodegradation and elastic modulus similar to that of human bone. The aim of our study is to develop a magnesium alloy with a controllable degradation that can closely match bone tissue to help injuries heal in vivo and avoid cytotoxicity caused by a sudden increase in ion concentration. In this study, we prepared and modified Mg-3 Zn, Mg-3Zn-1Y, and Mg-2Zn-1Mn by hot extrusion, and used Mg-2.5Y-2.5Nd was as a control. We then investigated the effect of additions of Y and Mn on alloys’ properties. Our results show that Mn and Y can improve not only compression strength but also corrosion resistance. The alloy Mg-2Zn-1Mn demonstrated good cytocompatibility in vitro, and for this reason we selected it for implantation in vivo. The degraded Mg-2Zn-1Mn implanted a bone defect area did not cause obvious rejection and inflammatory reaction, and the degradation products left no signs of damage to the heart, liver, kidney, or brain. Furthermore, we find that Mg-2Zn-1Mn can promote an osteoinductive response in vivo and the formation of bone regeneration.

Bone Regeneration in Osteoporosis via Carbon Nanotube-Based Bone Morphogenetic Protein-2

We constructed a bone morphogenetic protein 2 (BMP-2)@Carbon nanotube (CNT) delivery system to explore the feasibility of a nanodrug delivery system in the treatment of osteoporosis (OP). Osteoblasts were cultured and OP mouse models were constructed to evaluate the osteogenesis of nano-BMP-2 in OP therapy. In physicochemical property tests, we found that BMP-2 was effectively loaded into CNT to form nanoparticles (NPs) with a particle size of 100 nm. Additionally, we found that nano-BMP-2 had good stability and could effectively prolong BMP-2 release time. In cellular experiments, we found that nano-BMP-2 could penetrate osteoblasts more effectively than BMP-2 alone, and with the increase of BMP-2 loading, the amount of BMP-2 penetrating osteoblasts increased with an optimal loading of 100 μg. We determined that nano-BMP-2 could increase proliferation activity of osteoblasts to better promote OP repair. In our vivo experiments, we found that nano-BMP-2 was effectively excreted through the kidney and mainly distributed in bone tissue. Moreover, CNT effectively prolonged the half-life of BMP-2 and was safe to introduce through intramuscular injection and did not cause obvious inflammatory reactions. Following treatment, nano-BMP-2 increased body weight, femur weight, and femoral head diameter in OP mouse models. Furthermore, bone trabecular was arranged in a close and orderly fashion and was uniform in thickness in OP mice treated with nano-BMP-2. OP mice had improved bone mineral density, trabecular thickness, trabecular number, and cortical bone thickness in their metaphyseal regions, implying nano-BMP-2 treatment led to improved OP symptoms. Therefore, BMP-2@CNT may be a beneficial choice for treatment of OP.

Human Umbilical Cord Mesenchymal Stem Cells Inhibit Coronary Artery Injury in Nice with Lactobacillus casei Wall Extract-Induced Kawasaki Disease

Kawasaki disease (KD) is a serious threat to children’s physical and mental health. This study investigated the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on KD coronary arteritis induced by Lactobacillus casei wall extract (LCWE) in an animal model. Sixty BALB/C mice were randomly assigned to three groups (n = 20 mice per group). Mice in the model and stem-cell groups were injected with LCWE, while the control-group mice were injected with phosphate-buffered saline (PBS) for 2 days. At day 16 of modeling, PBS was injected into the control and model-group mice, and hUC-MSCs were injected into the stem-cell group mice for 10 days. At days 4, 15, 26, and 32 of modeling, echocardiography and histopathology were performed to examine the cardiac structure and the morphological changes in the coronary arteries in each group. B-ultrasonography showed that 57.5% (23/40) of the mice had coronary artery lesions, of which 5% (2/40) had right coronary artery aneurysm, 27.5% (11/40) had coronary artery wall thickening, a widened inner diameter of the main artery of the left coronary artery, and thickened intima. Histopathology showed slight swelling of the epicardium of the aortic valve, mitral valve, right ventricle, and atrium, as well as scattered infiltration of a few neutrophils. Following hUC-MSCs intervention treatment, B-ultrasonography showed a decrease in the main coronary artery diameter, while histopathology showed no obvious vascular inflammatory reaction or other obvious abnormalities. These findings highlight that hUC-MSCs inhibit coronary artery injury in animal models of KD induced by LCWE.

A Novel Drug Self-Delivery System from Fatty Alcohol Esters of Tranexamic Acid for Venous Malformation Sclerotherapy.

Venous malformation (VM), which causes severe damage to patients’ appearance and organ function, is one of the most common vascular malformations. At present, many drugs in clinical treatment cause various adverse reactions. Herein, we synthesized cationic amphiphilic gelators (TA6, TA8, and TA9) by introducing saturated carbon chains of different lengths to tranexamic acid (TA), which could self-assemble into low-molecular-weight gels (LMWGs) as drug delivery carriers by hydrogen bonds, van der Waals forces, and hydrophobic interactions. The rheological properties, gelation driving force and drug release profiles of TA6, TA8, and TA9 hydrogels were characterized, and the results indicated that the hydrogels prepared in this study possessed the typical characteristics of a gel and could release drugs slowly. More importantly, the TA9 gelator showed significant pharmacological activity, in that it served as both an active drug compound and a drug carrier. The in vitro experiments demonstrated that TA9 induced HUVECs death and hemolysis by destroying cell membranes in a dose-dependent manner, and caused cell death and hemolysis at a concentration of 0.09 µM/mL. Meanwhile, we found TA9 could interact not only with fibrinogen, but also with other endogenous molecules in the blood. After the administration of TA9 hydrogel for 15 days, macroscopic imaging and histological evaluation in mice and rabbits displayed obvious thrombi, inflammatory reactions, and venous embolization, indicating that the mechanism of the TA9 hydrogel in treating VM was involved in two processes. Firstly, the TA9 hydrogel relied on its mechanical strength to physically block veins and continuously release TA9, in situ, for targeted therapy. Then, TA9 destroyed endothelial cells and damaged venous walls critically, causing thrombi. Most excitingly, TA9 was hydrolyzed to TA by enzymes that inhibited the degradation of thrombi by plasmin to prolong the embolization time and to promote venous fibrosis. Compared with other clinically available sclerosants, the degradation of TA9 also empowered a better biocompatibility and biodegradability for the TA9 hydrogel. In conclusion, we synthesized a potentially safe and effective derivative of TA and developed a low-molecular-weight gel as a self-delivery system for TA in treating VM.

In Vitro and In Vivo Degradation, Mechanical Properties, and Histocompatibility of Weft-Knitted Silk Mesh-Like Grafts

To investigate the degradation, mechanical properties, and histocompatibility of weft-knitted silk mesh-like grafts, we carried out the In Vitro and In Vivo silk grafts degradation assay. The In Vitro degradation experiment was performed by immersing the silk grafts in simulated body fluid for 1 year, and the results showed that the degradation rate of the silk mesh-like grafts was very slow, and there were few changes in the mechanical properties and quality of the silk mesh-like graft. In Vivo degradation assay was taken by implantation of the silk mesh-like grafts into the subcutaneous muscles of rabbits. At 3, 6, and 12 months postoperation, the rate of mass loss was 19.36%, 31.84%, and 58.77%, respectively, and the maximum load was 63.85%, 34.63%, and 10.76%, respectively of that prior to degradation. The results showed that the degradation rate of the silk graft and the loss of mechanical properties In Vivo were faster than the results obtained in the In Vitro experiments. In addition, there were no significant differences in secretion of serum IL-6 and TNF-α between the experimental and normal rabbits (P >0.05), suggesting no obvious inflammatory reaction. The findings suggest that the weft-knitted silk mesh-like grafts have good mechanical properties, histocompatibility, and In Vivo degradation rate, and therefore represent a candidate material for artificial ligament

Clinical characteristics of human adenovirus infection in hospitalized children with acute respiratory infection in Beijing

Objective: To compare the clinical characteristics of different types of human adenovirus (HAdV) infection in hospitalized children with acute respiratory infection in Beijing, and to clarify the clinical necessity of adenovirus typing. Methods: In a cross-sectional study, 9 022 respiratory tract specimens collected from hospitalized children with acute respiratory infection from November 2017 to October 2019 in Affiliated Children's Hospital, Capital Institute of Pediatrics were screened for HAdV by direct immunofluorescence (DFA) and (or) nucleic acid detection. Then the Penton base, Hexon and Fiber gene of HAdV were amplified from HAdV positive specimens to confirm their HAdV types by phylogenetic tree construction. Clinical data such as laboratory results and imaging data were analyzed for children with predominate type HAdV infection using t, U, or χ2 test. Results: There were 392 cases (4.34%) positive for HAdV among 9 022 specimens from hospitalized children with acute respiratory infection. Among those 205 cases who were successfully typed, 131 were male and 74 were female, age of 22.6 (6.7, 52.5) months,102 cases (49.76%) were positive for HAdV-3 and 86 cases (41.95%), HAdV-7, respectively, while 17 cases were confirmed as HAdV-1, 2, 4, 6, 14 or 21. In comparison of clinical characteristics between the predominate HAdV type 7 and 3 infection, significant differences were shown in proportions of children with wheezing (10 cases (11.63%) vs. 25 cases (24.51%)), white blood cell count >15 ×109/L (4 cases (4.65%) vs.14 cases (13.73%)), white blood cell count <5×109/L (26 cases (30.23%) vs.11 cases (10.78%)), procalcitonin level>0.5 mg/L (43 cases (50.00%) vs. 29 cases (28.43%)), multilobar infiltration (45 cases (52.33%) vs.38 cases (37.25%)), pleural effusion (23 cases (26.74%) vs. 10 cases (9.80%)), and severe adenovirus pneumonia (7 cases (8.14%) vs. 2 cases (1.96%)) with χ²=5.11, 4.44, 11.16, 9.19, 4.30, 9.25, 3.91 and P=0.024, 0.035, 0.001, 0.002, 0.038, 0.002, 0.048, respectively, and also in length of hospital stay (11 (8, 15) vs. 7 (5, 13) d, Z=3.73, P<0.001). Conclusions: HAdV-3 and 7 were the predominate types of HAdV infection in hospitalized children with acute respiratory tract infection in Beijing. Compared with HAdV-3 infection, HAdV-7 infection caused more obvious inflammatory reaction, more severe pulmonary symptoms, longer length of hospital stay, suggesting the clinical necessity of further typing of HAdVs.

Acupuncture combined with smokeless or smoky moxibustion for regulating immune function of experimental chronic rhinosinusitis mice

To observe the therapeutic effect of moxa fume in the treatment of chronic rhinosinusitis（CRS） and the effect of acupuncture plus smokeless moxibustion or smoky moxibustion on the expression of thymic stromal lymphopoietin (TSLP) and pituitary adenylate cyclase activating polypeptide (PACAP) proteins in the sinus mucosal tissue in CRS mice. Sixty male C57BL/6J mice were randomly divided into 6 groups, namely normal control, sham operation, CRS model, medication, acupuncture plus smokeless moxibustion (Acu+smokeless Moxi) and acupuncture plus smoky moxibustion (Acu+smoky Moxi) groups, with 20 mice in each group. The CRS model was established by inserting a piece of polyporous sponge filled with streptococcus pneumoniae into the maxillary sinus after operation. The mice in the sham operation group received skin incision after opening the maxillary sinus. Mice of the medication group received gavage of clarithromycin 0.103 g·kg-1·d-1 for 21 days. For mice of the Acu+smokeless Moxi and Acu +smoky Moxi groups, manual acupuncture stimulation was applied to bilateral "Zusanli" (ST36), "Shenshu" (BL23) and "Hegu" (LI4) with the needles retained for 30 min, once every other day, and on the following day, moxibustion was applied to "Guanyuan" (CV4) and "Shenque" (BL23) for 20 min, once every other day. The treatment was given for 21 days. Mice of the normal, sham operation and model groups received gavage of normal saline (200 μL/d) for 21 days. Histopathological changes of the nasal mucosa were observed after H.E. staining, the TSLP and PACAP contents and expression were determined by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, separately. At the end of the treatment, mice of the model group still had symptoms of nasal obstruction and runny nose, but those of the 3 treatment groups were obviously relieved in the nasal symptoms. H.E. staining showed an obvious chronic inflammatory reaction in the sinus mucosa, uneven distribution of the mucosal epithelium and necrotic and exfoliated epithelial cells, hyperplasia of fibrous tissue in the submucosa, etc. in the model group, which were relatively milder in the medication, Acu+smokeless Moxi and Acu+smoky Moxi groups, while no obvious inflammation was found in the normal group and sham operation group. In comparison with the normal group, no significant changes were found in the expression levels of PACAP and TSLP in the sham operation group (P>0.05). The expression level of PACAP was significantly lower (P<0.05) and that of TSLP significantly higher in the model group than in the normal and sham operatin groups (P<0.05). Compared with the model group, no significant changes were found in the expression of PACAP in the medication, Acu+smokeless Moxi and Acu+smoky Moxi groups (P>0.05), and the expression of TSLP was further obviously increased in the Acu+smokeless Moxi group (P<0.01), but obviously decreased in the Acu+smoky Moxi group (P<0.01). Acupuncture combined with smoky moxibustion can down-regulate the expression of TSLP protein in the nasal sinus mucosa in CRS mice, which maybe contribute to its effect in reducing the inflammatory reaction and nasal symptoms.

Decellularized tendon matrix membranes prevent post-surgical tendon adhesion and promote functional repair

Adhesion often occurs after tendon injury, and results in sliding disorder and movement limitation with no ideal solution for it in clinic. In this study, an anti-adhesion membrane, i.e., decellularized tendon matrix (DTM) for tendon is successfully prepared by an optimized tendon decellularization method from homologous extracellular matrix. Microsection technology has been used to optimize the method of decellularization in order to better preserve the bioactive components in tissues and reduce the chemical reagent residues on the premise of effective decellularization with relatively shorter time and less reagents for decellularization. The physic-chemical properties and biological functions of DTM are evaluated, and high-throughput and high-precision tandem mass tags (TMT) labeling proteomics technology is used to analyze protein components of DTM, which may provide the scientific support for application of the innovative product. In vitro biosafety tests show that DTM not only is non-toxic but also promote cell proliferation. Subcutaneous implantation test confirms that DTM is completely degraded after 12 weeks and there is no obvious inflammatory reaction. The results of Achilles tendon repair in rabbits show that DTM can not only prevent tendon adhesion but also improve the quality of tendon repair, which demonstrates its tremendous application potential. Statement of SignificanceThere is no ideal solution for adhesion after tendon injury. In this study, a dense tendon anti-adhesion membrane (DTM) was successfully prepared from homologous extracellular matrix (ECM). This DTM could effectively retain bioactive ingredients, and prevent adhesion as well as improve the quality of tendon repair in vivo. An optimized decellularization method was used which could effectively decellularize tendon in a short time, better preserve bioactive components, and reduce reagent residues. For the first time, high-throughput and high-precision tandem mass tags (TMT) labeling proteomics technology was used to qualitatively and quantitatively analyze the protein composition of fresh tendon, acellular tendon and DTM, which provided not only scientific support for the application of DTM, but also comprehensive and accurate data support for related research of bovine tendons and decellularization.

Comparative Evaluation of Neuropathological Lesions Progression in Different Brain Areas in A Rat Model of Chronic Hepatic Encephalopathy

Hepatic encephalopathy (HE) is considered a serious clinical complication of acute and chronic liver injury. Previous researches elucidated HE but the characterization of neuropathology in different brain areas is limited. The present study is conducted to evaluate the progression of neuropathological lesions in different brain areas in a rat model with liver fibrosis induced by thioacetamide (TAA). For this experiment, 20 male rats were divided into two groups control and TAA-treated group that was I.P injected twice a week with 100 mg/kg.b.w TAA dissolved in saline for 3 months. Three rats from each group were euthanized after 3 months and the liver specimens were collected for ensuring the development of a rat model for liver fibrosis. The remained rats were kept without further treatment with TAA for 2 months, rats were carefully monitored for abnormal signs. After the end of the experimental period, rats were euthanized and the brains were collected for histopathological examination. Microscopic examination of the brain revealed various neuropathological lesions that varied in severity between different brain areas. The most severe lesions were recorded in the midbrain and pons while mild lesions were detected in the cerebral cortex and striatum. The brain lesions comprised varied degrees of neuronal degeneration with marked astroglial and microglial reactions in the different brain regions. The obvious inflammatory reaction indicates that neuroinflammation plays an important role in the mediation of HE.

ВЫЯВЛЕНИЕ УСТОЙЧИВОСТИ К АНТИБИОТИКАМ У ВОЗБУДИТЕЛЯ ГИСТОФИЛЕЗА КРУПНОГО РОГАТОГО СКОТА Histophilus somni

ВЫЯВЛЕНИЕ УСТОЙЧИВОСТИ К АНТИБИОТИКАМ У ВОЗБУДИТЕЛЯ ГИСТОФИЛЕЗА КРУПНОГО РОГАТОГО СКОТА Histophilus somni

PROPERTIES OF EXPERIMENTAL SAMPLES OF VACCINE AGAINST AVIAN INFECTIOUS CORYZA

PROPERTIES OF EXPERIMENTAL SAMPLES OF VACCINE AGAINST AVIAN INFECTIOUS CORYZA

Esophagitis dissecans superficialis associated with acute transoral paraquat poisoning: Clinical study of 15 cases.

To analyze the clinical characteristics and therapeutic effects of transoral paraquat poisoning combined with Esophagitis dissecans superficialis (EDS). A retrospective observational study was conducted on paraquat poisoning patients between January 1, 2011 and August 30, 2016 in Qilu hospital. Fifteen patients with EDS were enrolled in this study. The clinical characteristics, prognosis, and pathological features of esophageal necrosis mucosa of these patients were retrospectively analyzed and summarized. Esophageal mucosal dissection occurs mainly within 3-8 days after transoral paraquat poisoning in 15 patients. Dosage of paraquat is range from 50 to 100 ml. Most patients have physical problems with swallowing before the intramural esophageal dissection occurred. And there are other symptoms, including sore throat or dysphagia (100%), nausea and vomiting (86.7%), heartburn or upper abdominal pain (73.3%), hematemesis (60%), abdominal distension (20%) and cough frequently (6.7%). In death group, most patients demonstrate features of the multiple organ failure when the esophageal mucosal stripping happened, including lung injury, renal failure, and hepatic failure. The shape of esophageal dissection was tubular in 60%, irregular in 40%, and they vary in size. Pathological examination showed extensive injury, necrosis and hemorrhage of digestive tract epithelium, and obvious inflammatory reaction of epithelial tissue. Transoral paraquat poisoning has certain damage to the patient's esophageal mucosa, and some may be complicated with EDS, and the prognosis is poor, especially when combined with multiple organ dysfunction. Esophageal damage is mainly located in the esophageal mucosa and have different degrees. Special attention should be paid on such patients.

Effects of Umbilical Cord Blood Stem Cells on Coronary Artery Damage in Mice with Kawasaki Disease By High-Frequency Ultrasound

Objective: to use lactobacillus casein cell wall extract (LCWE) to induce Kawasaki disease (KD) mouse model, and analyse the process of coronary artery damage and effects of umbilical cord blood stem cells by high-frequency ultrasound in small animals. Methods: LCWE was prepared and 18 BALB/c pups were randomly divided into two groups: 15 in KD model group and 3 in the normal control group. KD model was induced by a single intraperitoneal injection of 0.5ml LCWE in the model group, and changes of the coronary artery were observed at 2d, 21d and 30d after the injection, respectively. From the 16th day of modeling, 300 μL PBS was injected intraperitoneally daily in the control group and model group. The hUC-MSCs 300 μL (105/mL) were intraperitoneally injected daily for 10 consecutive days in the stem cell group. The mice were sacrificed in batches on day 2, 15, 21 and 30, and the morphological changes of coronary arteries were observed by echocardiography and histopathology. Results: the change of coronary artery diameter could be accurately measured by high-frequency small animal ultrasound. At 21d, the coronary arteries of the model group were widened compared with those of the control group. At 30d, there was no significant difference between the model group and the previous model group. Histopathology showed slight swelling of the epicardium of aortic valve, mitral valve, right ventricle and atrium, scattered infiltration of a few neutrophils, dilatation of the coronary artery lumen, necrosis and disintegration of a small number of myocardial cells, and significant hyperplasia of local fibrous connective tissue accompanied by solid calcium salt deposits. After hUC-MSCs intervention treatment, B-ultrasound showed a decrease in the main coronary artery diameter, histopathology showed multiple lymphocytes, eosinophilic granulocytes, and mononuclear cell infiltration in the left atrial appendage of mice, and no obvious vascular inflammatory reaction or other obvious abnormalities were observed. Conclusion: high-frequency ultrasound can be used to clearly obtain the coronary artery image of KD mice and dynamically observe the evolution process of coronary artery diameter, which provides more diagnostic basis for the treatment of clinical KD. And hUC-MSCs intervention reduced the pathological lesion of coronary artery inflammation in mice compared with the model group.

Lentiviral vector-mediated expression of C3 transferase attenuates retinal ischemia and reperfusion injury in rats

AimsOur previous study showed that intravitreal delivery of self-complementary AAV2 (scAAV2)-mediated exoenzyme C3 transferase (C3) can attenuate retinal ischemia/reperfusion (I/R) injury. The current study investigated the neuroprotective effects of lentivirus (LV)-mediated C3 transgene expression on rat retinal I/R injury. Main methodsThe LV encoding C3 and green fluorescent protein (GFP) together (LV-C3-GFP) or GFP only (LV-GFP) was intravitreally injected to SPRAGUE-DAWLEY rats. On day 5 post-intravitreal injection, eyes were evaluated by slit-lamp examination. The GFP expression on retina was confirmed by in vivo and ex vivo assessments. RhoA GTPase expression in retina was examined by western blot. Retinal I/R injury was generated by transiently increasing intraocular pressure (110 mmHg, 90 min). Eyes were then enucleated, and retinas processed for morphological analysis and TdT-dUTP terminal nick-end labeling (TUNEL) assay. Key findingsNo obvious inflammatory reactions or surgical complications were observed after intravitreal injection of LV vectors. There was a significant decrease of total RhoA GTPase level in the retina treated with LV-C3-GFP. Compared to the blank control group, LV-C3-GFP and LV-GFP did not affect the retinal thickness, cell density in ganglion cell layer (GCL), or numbers of apoptotic cells in retinal flat-mounts. In the LV-GFP-treated retinas, I/R decreased the retinal thickness and GCL cell density and increased apoptotic retinal cell numbers. LV-C3-GFP significantly protected against all these degenerative effects of I/R. SignificanceThis study indicated that LV-mediated C3 transgene expression exhibits neuroprotective effects on the retinal I/R injury and holds potential as a novel neuroprotective approach targeting certain retinopathies.

Differences in the microcirculation disturbance in the right and left ventricles of neonatal rats with hypoxic pulmonary hypertension

Microcirculation disturbance is a crucial pathological basis of heart damage; however, microcirculation alterations induced by hypoxic pulmonary hypertension (HPH) remain unknown, and the left ventricle (LV) in HPH is conventionally ignored. Herein, we investigated the changes in the cardiac structure, function and microcirculation after HPH and further compared the differences between the right ventricle (RV) and LV. Using a neonatal rat model of HPH, we found RV myocardial hypertrophy, dysfunction and poor myocardial perfusion in HPH rats. Additionally, RV microcirculation disturbance manifested as the abnormal expression of endothelin-1/eNOS and increased expression of intercellular cell adhesion molecule-1 (ICAM-1) or E-selectin 3 days after hypoxia, followed by vascular inflammation, coronary arterial remodeling and microvascular sparseness. Impairment in LV vasodilation was detected in rats after 3 days of hypoxia; however, no obvious microvascular rarefaction or inflammatory reaction was observed in the LV. In conclusion, our results suggest that HPH mainly triggers RV microcirculation disturbances, causing low myocardial perfusion damage and cardiac dysfunction. Despite the differences in the RV and LV, their impaired microvascular function, mediated by endothelial cells, occurs almost simultaneously after HPH, earlier than cardiac functional or structural abnormalities.

Clinical significance and prognostic value of cellular immunity, inflammatory factors and myocardial enzymes in septic shock

Objective To investigate the clinical significance of cellular immune function, inflammatory factors and myocardial enzyme detection in evaluation of prognosis of septic shock, so as to provide a reference for clinical diagnosis and treatment of septic shock. Methods From June 2015 to June 2018, 73 patients with septic shock treated in the First People's Hospital of Yongkang were selected as the research subjects.According to the 28d prognosis after the patients entered the ICU, the patients were divided into the death group and the survival group.Another 67 cases of health examination from June 2015 to June 2018 were selected as control group.The changes of T lymphocyte subsets includingCD3, CD4 and CD8 were measured by BD FACSCanto I I flow cytometry, the content of procalcitonin(PCT) was determined by immunoluminescence assay, the content of C-reactive protein(CRP) was determined by immunoturbidimetry, and the content of creatine kinase MB(CK-MB), cardiac troponin I(cTnI) and lactate dehydrogenase(LDH) was determined by enzymatic kinetics. Results The APACHE Ⅱ score and MODS score of the survival group and the death group were higher than those of the control group [(2.20±0.61)points and (4.86±1.29)points]. The APACHE Ⅱ score and MODS score of death group [(16.45±4.28)points and (27.63±4.97)points] were higher than those of survival group [(9.84±2.45)points and (19.84±3.28)points](all P<0.05). The CD3+, CD4+ and CD4+/CD8+ in the survival group and the death group were significantly lower than those in the control group[(71.32±6.96)%, (42.63±4.26)%, (1.67±0.31)%](all P<0.05). The contents of PCT and CRP in the survival group and the death group were higher than those in the control group [(0.19±0.03)ng/mL and (2.19±0.76)mg/L], and the contents of PCT [(15.93±3.26)ng/mL] and CRP [(184.32±29.80)mg/L] in the death group were higher than those in the survival group [(6.87±1.94)ng/mL and (69.49±17.42)mg/L] (all P<0.05). The CK-MB content in the survival group and the death group of septic shock was lower than that in the control group [(1.97±0.21)μg/L], and the cTnI and LDH contents were higher than that in the control group [(0.03±0.01)μg/L and (168.93±16.52)U/L], the content of CK-MB [(0.68±0.10)μg/L] in the death group was lower than that in the survival group [(1.27±0.13)μg/L], while the contents of cTnI [(0.39±0.06)μg/L] and LDH [(384.52±39.89)U/L] in the death group were lower than those in the survival group [(0.17±0.04)μg/L and (257.18±25.47)U/L] (P<0.05). Conclusion The immune function of the patients with septic shock is obviously reduced, there are obvious inflammatory reactions and abnormal myocardial enzymes, and the detection of cellular immune function, inflammatory factors and myocardial enzymes can be used as an effective index to judge the shock of septic shock. Key words: Shock, septic; Immunity, cellular; C-reactive protein; Creatine kinase, MB form; Troponin I; Lactate dehydrogenases; Prognosis