Subcutaneous Injection Of Normal Saline Research Articles

Controlling redox state by edaravone at transplantation site enhances bone regeneration

In cell-based bone augmentation, transplanted cell dysfunction and apoptosis can occur due to oxidative stress caused by the overproduction of reactive oxygen species (ROS). Edaravone (EDA) is a potent free radical scavenger with potential medical applications. This study aimed to investigate the effect of controlling oxidative stress on bone regeneration using EDA. Bone marrow-derived cells were collected from 4-week-old rats, and EDA effects on cell viability and osteogenic differentiation were evaluated. Collagen gels containing PKH26-prelabeled cells were implanted into the calvarial defects of 12-week-old rats, followed by daily subcutaneous injections of normal saline or 500 μM EDA for 4 d. Bone formation was examined using micro-computed tomography and histological staining. Immunofluorescence staining was performed for markers of oxidative stress, macrophages, osteogenesis, and angiogenesis. EDA suppressed ROS production and hydrogen peroxide-induced apoptosis, recovering cell viability and osteoblast differentiation. EDA treatment in vivo increased new bone formation. EDA induced the transition of the macrophage population toward the M2 phenotype. The EDA group also exhibited stronger immunofluorescence for vascular endothelial growth factor and CD31. In addition, more PKH26-positive and PKH26-osteocalcin-double-positive cells were observed in the EDA group, indicating that transplanted cell survival was prolonged, and they differentiated into bone-forming cells. This could be attributed to oxidative stress suppression at the transplantation site by EDA. Collectively, local administration using EDA facilitates bone regeneration by improving the local environment and angiogenesis, prolonging survival, and enhancing the osteogenic capabilities of transplanted cells.

Anise (Pimpinella anisum L.) attenuates azoxymethane-induced colorectal cancer by antioxidant, anti-inflammatory, and anti-apoptotic pathways in rats.

Herbal medicine is one of the most common fields explored for combating colon cancers, and Pimpinella anisum L. seeds (PAS) have been utilized widely as medicinal agents because of their increased essential oil (trans-anethole) contents. In this essence, our study investigates the toxic effect and chemoprotective potentials of PAS against azoxymethane (AOM)-induced colon cancer in rats. The toxicity trial for PAS conducted by clustering fifteen rats into three groups (five rats each): A, normal control had 10% Tween 20; B, ingested with 2g/kg PAS; and C, supplemented with 4g/kg PAS. The in vivo cancer trial was performed by using 30 rats (Sprague-Dawley) that were randomly adapted in five steel cages (six rats each): group A, normal controls received two subcutaneous injections of normal saline 0.09% and ingested orally 10% Tween 20; groups B-E, rats received two injections of 15mg/kg of azoxymethane (AOM) subcutaneously in 2weeks and treated orally with 10% Tween 20 (group B) or intraperitoneal injection of 5-fluorouracil (35mg/kg) (group C), or orally given 200mg/kg PAS (group D) and 400mg/kg PAS (group E) for 8weeks. After the scarification of rats, the colon tissues were dissected for gross and histopathological evaluations. The acute toxicity trial showed the absence of any toxic signs in rats even after 14days of ingesting 4g/kg of PAS. The chemoprotective experiment revealed significant inhibitory potentials (65.93%) of PAS (400mg/kg) against aberrant crypto foci incidence that could be correlated with its positive modulation of the immunohistochemically proteins represented by a significant up-regulation of the Bax protein and a decrease of the Bcl-2 protein expressions in colon tissues. Furthermore, PAS-treated rats had notably lower oxidative stress in colon tissues evidenced by decreased MDA levels and increased antiradical defense enzymes (SOD, CAT, and GPx). The outcomes suggest 400mg/kg PAS as a viable additive for the development of potential pharmaceuticals against colorectal cancer.

Umbilical Cord Wharton's Jelly-Derived Mesenchymal Stem Cells Inhibit the TGF-β1 Pathway in Hepatic Fibrosis Rats Through a Paracrine Regulation Process

The aim is to verify the therapeutic effect and possible mechanism of human umbilical cord Wharton's jelly-derived transplantation of mesenchymal stem cells (UMSCs) on CCl4-induced hepatic fibrosis rats through in vivo studies and to explore the regulatory mechanism of UMSCs on fibrosis of hepatic stellate cells (HSCs) through in vitro experiments. In vivo experiment: Rats were randomly divided into blank control group and hepatic fibrosis group. During the entire trial, the blank control group received subcutaneous injection of normal saline, while in the hepatic fibrosis group received injections of 50% CCl4-olive oil subcutaneously for 10 weeks to establish the rat model of liver fibrosis. Hepatic fibrosis rats were then randomly and evenly divided into umbilical cord mesenchymal stem cell (UMSC) group, bone marrow mesenchymal stem cell (BMSC) group, UMSC-culture medium (CM) group, and control group. Rats in each group were infused with the following substances through the caudal vein as follows: 1 mL UMSCs (2 × 106/mL) in UMSC group, 1 mL BMSCs (2 × 106/mL) in BMSC group, 1 mL UMSCs-CM in CM group, and 1 mL saline in control group. Rats of each group were closely observed (weight, hair condition, activity, appetite, diarrhea, etc.), venous blood samples were collected, the number of white blood cells and lymphocytes were measured, and liver function indicators (ALT, AST, TBIL, ALB) were determined. Three weeks later, rat liver specimens were taken, HE stained, pathological changes were examined and quantified. In vitro experiments: HSCs were seeded in 6-well plates at 1.0 × 105/mL, with a serum-free medium for 24 hours. Then, 2 mL of UMSCs-CM was added in the study group, while an equal amount of complete medium was added to the control group. RT-PCR was used to detect TGF-β1, Collagen-I, TIMP-2 mRNA expression in HSCs, and western blot was used to detect TGF-β1 protein expression in HSCs. In vivo experiment: Compared with the control group, after the transplantation, the activity status (weight, spirit, appetite, movement, hair, diarrhea, etc.) of rats in the UMSC group, BMSC group, and CM group were improved. The liver function indexes of these groups, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (TBIL) were significantly decreased (p < 0.05), while albumin (ALB) levels were mildly but not significantly increased (p > 0.05). The Knodell score (reflecting the degree of liver inflammation) and Chevallier score (reflecting the degree of liver fibrosis) of liver specimens in pathological examination were also significantly reduced, and the difference in the quantitative scores of those indexes was statistically significant (p < 0.05). There was no statistically significant difference in the number of venous white blood cells and lymphocytes, liver function indexes (ALT, AST, TBIL, ALB), Knodell score, and Chevallier score of liver samples among the UMSC group, BMSC group, and CM group. In vitro experiments: After treatment with UMSCs-CM, the expression of TGF-β1, Collagen-I, and TIMP-2 mRNA in HSCs was significantly down-regulated compared with that of the control group (treated with complete medium), and it gradually decreased with the extension of the treatment time. Compared with the control group, the expression of TGF-β1 protein in the HSCs of the experimental group was down-regulated, and this effect was time-dependent, specifically, the control group (2.49 ± 0.43) > the experimental group at 48 hours (1.98 ± 0.26) > the experimental group at 72 hours (1.62 ± 0.20) (F = 7.796, p < 0.05). In rats with liver fibrosis, transplantation of UMSCs can improve liver function and reduce the inflammatory activity and fibrosis of the liver, possibly through the paracrine mechanism. UMSCs inhibit HSCs fibrosis through a paracrine mechanism, which is time-dependent, possibly by targeting TGF-β1 and its downstream gene products.

The effects of crude alkaloid extract of Matricaria chamomilla L. on convulsions induced by pentylenetetrazole in chicks

The present research was designed to evaluate the protective effect of crude alkaloid extract of Matricaria chamomilla L. (MC) against pentylenetetrazole (PTZ) induced convulsions in chicks. A total of seventy-two chicks were randomly divided into six groups each one consisting of 12 chicks. The first group was considered to be a negative group that received subcutaneous injection of normal saline, the second group (positive control) received subcutaneous injection of PTZ (90 mg/kg), and the third group received sodium valproate (200 mg/kg) orally for six days. The other groups (fourth, fifth and sixth) were treated with single intraperitoneal injection of crude alkaloids extract of MC (20, 40, 80 mg/kg) respectively. After 30 minutes of the treatment, the groups (third to sixth) received PTZ and observed the convulsion signs for the next 30 minutes. At the end of the experiment, various biochemical parameters were assessed including, brain gamma-aminobutyric acid (GABA) and glutamate, serum catalase (CAT), glutathione reductase (GR), malondialdehyde (MDA), and 8-isoprostane, and some electrolytes (potassium K+, sodium Na+, chloride Cl-, ionized calcium iCa2+, total calcium TCa2+), pH, and glucose levels in serum. The results indicated that the group treated with crude alkaloids extract at higher dose (80 mg/kg) showed a significant increase in CAT and significantly decreased in 8-isoprostane, and an increase in Na+ ions concentration in serum. It was concluded that pretreatment of crude alkaloids extract (80 mg/kg) of MC offers anticonvulsant activity by reducing the mortality rate induced by PTZ, as well as it plays an important role in inhibiting oxidative stress.

An in vivo rabbit joint injury model to measure trauma-induced coagulopathy and the effect of timing of administration of ketotifen fumarate on posttraumatic joint contracture.

Objectives:Using a rabbit in vivo joint injury model, the primary objective of the study was to determine if a relationship exists between earlier time to initiation of ketotifen fumarate (KF) treatment and posttraumatic joint contracture (PTJC) reduction. The secondary objective was to determine if a coagulation response could be detected with serial thrombelastography (TEG) analysis following acute trauma in this model.Methods:PTJC of the knee were created in 25 skeletally mature, New Zealand White rabbits. Five groups of 5 animals were studied: a control group that received twice daily subcutaneous injections of normal saline and 4 treatment groups that received twice daily subcutaneous injections of KF (0.5 mg/kg) starting immediately, 1-, 2-, and 4-weeks post-injury. After 8 weeks of immobilization, flexion contractures were measured biomechanically. Serial TEG analysis was performed on the control group animals pre-injury and weekly post-injury.Results:The average joint contracture in the Control Group (43.1° ± 16.2°) was higher than all KF treatment groups; however, the differences were not statistically significant. The average joint contracture was lowest in the 2-week post-injury treatment group (29.4° ± 12.1°), although not statistically significant compared to the other treatment groups. Serial TEG analysis demonstrated significantly higher mean maximal amplitude (maximal amplitude = 68.9 ± 1.7 mm; P < .001), alpha-angle (81.9° ± 0.9°; P < .001), and coagulation index (4.5 ± 0.3; P < .001) 1-week post-injury, which normalized to pre-injury values by 5-weeks post-injury.Conclusions:The use of the mast cell stabilizer KF within 2 weeks of injury demonstrated a nonsignificant trend towards reducing joint contracture in a rabbit in vivo model of PTJC. TEG and the in vivo rabbit joint injury model may be valuable in future preclinical studies of venous thromboembolism prevention and furthering our understanding of the pathophysiology of posttraumatic hypercoagulability.

Analgesia for major laparoscopic abdominal surgery: a randomised feasibility trial using intrathecal morphine

Effective pain control enhances patient recovery after surgery. Laparoscopic techniques for major abdominal surgery are increasingly utilised to reduce surgical trauma. Intrathecal morphine is an attractive analgesic option that is gaining popularity. However, limited evidence guides its use in the setting of laparoscopic surgery. In addition, enhanced recovery after surgery pathways advocate opioid-sparing techniques. We conducted a feasibility trial to compare intrathecal morphine with non-neuraxial analgesia in laparoscopic or laparoscopic-assisted major abdominal surgery to inform the design of a future large clinical trial. This multicentre, double-blind, randomised controlled trial was conducted at two tertiary hospitals in Australia. Fifty-one patients were randomly allocated to receive either intrathecal morphine (intervention group) or a sham subcutaneous injection of normal saline in the lumbar area (control group) immediately before the induction of general anaesthesia. Co-primary outcomes were patient recruitment and successful adherence to treatment allocation as per the study protocol. The primary endpoints of feasibility and protocol adherence were met with a 46% recruitment rate (51 of 110 eligible patients) and 96% protocol adherence. There was only one patient with failed access to the intrathecal space. For secondary endpoints, fewer patients in the intrathecal morphine group required opioids in the post-anaesthesia care unit, their postoperative pain scores at rest were lower across the four time-points measured (p = 0.007), but not dynamic pain scores (p = 0.061), and pruritus was more common following intrathecal morphine (p = 0.007). Total oral morphine equivalents until postoperative day 3 were less in the intrathecal morphine group (median (95%CI) difference 82 (-13 to 168) mg), but this reduction was not statistically significant (p = 0.10). These findings support conducting a definitive clinical trial.

Study on Toll-Like Receptor 2-Mediated Inflammation-Induced Familial Hypertension Combined with Hyperlipemia and Its Mechanism.

According to the latest clinical data, cardiovascular diseases have ranked first in prone diseases, causing 40% of the premature deaths of China's population. This study aimed to investigate the influence of Toll-like receptor 2- (TLR2-) mediated inflammation on the occurrence and development of familial hypertension combined with hyperlipemia and its related mechanism. Blood specimens from 66 patients undergoing coronary atherosclerosis were collected and grouped, including 22 patients into the control group, 25 into the familial hypertension group, and 19 into familial hypertension combined with hyperlipemia group. In this study, ELISA was conducted for determining the levels of four inflammatory factors of TLR2 and IL-1β, IL-6, TNF-ɑ, and CCL2 in serum and the levels of relevant indicators in mice. C57Bl/6j and genetically engineered C.129(B6)-Tlr2tm1Kir/J mice were given subcutaneous injection of normal saline (wild-saline group), 8-week 40% high-fat diet (wild-high-fat group), and subcutaneous Alzet-implanted angiotensin II micropump supplemented with the research diet (wild-high fat-Ang II group, Tlr2-/--high fat-Ang II group). Blood pressure in mice was recorded consecutively with a noninvasive hemopiezometer for eight weeks. TLR2 and IL-1β, IL-6, TNF-ɑ, and CCL2 in serum of patients with familial hypertension combined with hyperlipemia and the hypertension combined with hyperlipemia mouse model were higher than those in the normal group. Under combined intervention of Ang II and the research diet, mRNA expression related to blood pressure, blood lipid, and fat metabolism in Tlr2-/- genetically engineering mice was significantly lower than that in the wild-high fat-Ang II group. The phosphorylation levels of AKT, IKK, and p65 in mice with hypertension combined with hyperlipidemia were significantly higher than those in normal group. The levels of blood pressure and blood lipid in mice after blocking the AKT or NF-κB pathway were significantly downregulated compared with those in the wild-high fat-Ang II group, with statistically significant differences (both P < 0.05). In conclusion, TLR2 regulates inflammation through Akt-NF-κB pathway, thus inducing the occurrence and development of familial hypertension combined with hyperlipemia.

SO076EVOLOCUMAB HAS THERAPEUTIC EFFECTS ON EXPERIMENTAL MODEL OF FSGS

Abstract Background and Aims Proprotein convertase subtilisin/kexin type 9 (PCSK9), which accelerates LDL-receptor degradation, plays a central role in dyslipidemia in nephrotic syndrome (NS). Intracellular LDL-C accumulation is supposed to damage podocytes through oxidative stress, foam cell formation, and apoptosis. In the present study, we investigate the effects of evolocumab (EVO), which is anti-PCSK9 monoclonal antibody, on a murine model of adriamycin-induced focal segmental glomerulosclerosis (FSGS) with NS in order to clarify the potential of EVO as a therapeutic agent in FSGS. Method Male BALB/c mice aged 9 to 11 weeks were divided into vehicle and intervention groups. The mice underwent subcutaneous injection of normal saline (FSGS-Vehicle group) or EVO (30 mg/kg) (FSGS-EVO group) and immediately after that, they were administered adriamycin (11.5 mg/kg) from tail vein. All mice were sacrificed on Day 14, and then morphological and functional analyses were performed. Results EVO treatment significantly reduced serum levels of LDL-C (mg/dl, 305.3±77.1 vs. 159.1±31.7, p&amp;lt;0.01) and PCSK9 (ng/ml, 652.9±230.0 vs 311±116.5, p&amp;lt;0.001). In addition, EVO treatment significantly improved renal function, including albuminuria (albumin/creatinine (Cr), 15.73±1.26 vs. 7.32±1.02, p&amp;lt;0.001), serum Cr (mg/dl, 0.48±0.13 vs. 0.18±0.044, p&amp;lt;0.001) and BUN (mg/dl, 80.08±20.22 vs. 46.86±9.60, p&amp;lt;0.05). Compatibly with the clinical data, the severity of glomerulosclerosis score (semi-quantification, 2.44 vs. 1.95, p&amp;lt;0.001) was ameliorated and podocyte density (number of podocyte/glomerulus, 2.61 vs. 7.54, p&amp;lt;0.001) was maintained in the FSGS-EVO mice compared with the FSGS-Vehicle mice. Conclusion Our findings provided novel insights into the protective effects of EVO on adriamycin nephropathy, indicating EVO as a potent therapeutic agent for FSGS.

Action mechanism of microRNA-1a-3p in mouse neurogenic bladder

Objective To establish a mouse model of neurogenic bladder and to investigate the changes of fibronectin 1 (FN1) and the action mechanism of microRNA (miRNA, miR)-1a-3p. Methods Mice were divided into model group and control group. The model group received subcutaneous injection of myelin oligodendrocyte glycoprotein and mycobacterium tuberculosis, and intraperitoneal injection of pertussis toxin at 48 h. The control group received subcutaneous injection of normal saline. The frequency of urination and the amount of urine output were observed. The mice were sacrificed and the bladder tissue was taken. Three bladders were selected from the model group and the control group for high-throughput sequencing. The changes of FN1 and miR-1a-3p in mouse bladder tissues were detected by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blotting. Results The sequencing results were analyzed. As the urinary system symptoms worsened, FN1 increased (4.569±0.426, t=-5.142, P<0.01), while miR-1a-3p decreased (1.623±0.312, t=-3.945, P<0.05). Compared with the control group, the expression of FN1 mRNA [2.501 (1.301, 4.251), F=99.183, P<0.01] and protein [2.937 (1.174, 4.262), F=63.834, P<0.01] in the bladder tissue of the model group was up-regulated, and that of miR-1a-3p was down-regulated [2.401 (1.250, 3.001), F=35.951, P<0.01]. The dual luciferase gene report indicated that FN1 was a direct target gene of miR-1a-3p (0.514±0.027, t=13.355, P<0.01). Conclusion The animal model of neurogenic bladder has urinary frequency, urgency, urinary retention and other symptoms, and the expression of FN1 is up-regulated, while the expression of miR-1a-3p is down-regulated. Therefore, miR-1a-3p may participate in neurogenic bladder formation through regulation of FN1. Key words: Neurogenic bladder; Fibronectin 1; MicroRNA-1a-3p

Effects of Nicotine Administration on the Structure of Auditory Cortex of Adolescent Male Guinea Pigs, a Histological and Ultrastructural Study

Background: Nicotine, the main ingredient in tobacco smoke, has always been linked to degenerative changes to the nervous system and several areas in the brain were reported to be injured due to nicotine. The effect of nicotine on the auditory system is only being recognized recently with few studies assessed the morphology. The effect of nicotine on the primary auditory cortex of young adolescent animals was addressed in this study.Materials and Methods: Twenty young male guinea pigs of two months old were divided into two groups of 10 animals each. Group I, the control group, received daily subcutaneous injections of normal saline for one month. Group II, the nicotine-treated group, received 3 mg/Kg body weight of nicotine subcutaneously daily. After animal sacrifice, brains were removed and processed for light and electron microscopic evaluations. Morphometry was also done to light microscopic histological sections.Results: In the nicotine-treated group, there were degenerative changes affecting the neurons, glia as well as blood capillaries. There was a darkening of neurons and disruption of their dendrites and organelles. The glial cells revealed reactivity, swelling, and cytoplasmic disruption. Blood capillaries showed collapse and thickening of their basement membrane. Morphometry revealed that the thickness of the auditory cortex has decreased as well as the dark neuronal number has increased in the treated group versus the control.Conclusion: Nicotine administration to adolescent male guinea pigs resulted in degenerative changes affecting the auditory cortex of the brain, which emphasizes the hazardous effects of cigarette smoking, especially at a young age.

Abatacept induced granulomatous hepatitis with a sarcoidosis- like reaction: a blinded trial in mice

BackgroundAbatacept is increasingly used for rheumatoid arthritis (RA) and juvenile idiophathic arthritis (JIA) treatment. However little is known about the risk of hepatotoxicity. The aim of this study was to determine whether the inhibition of the T cell CD28 receptor by abatacept results in acute hepatitis in BALB/c mice.MethodsTwenty BALB/c mice were studied. Ten mice received subcutaneous (SC) injection of abatacept (0.25mg per 25g body weight per 0.03 ml normal saline) at 0, 2, 4 and 8 weeks. For the control group, 10 mice received a SC injection of normal saline (NS) (0.03 ml). At the 10th week post injection, the mice were sacrificed, and histopathological studies were conducted.ResultsOf the abatacept-treated group, 3/10 mice died. Liver histology for the abatacept-treated group showed that 6/7 displayed histopathological changes in the lobular cellular infiltrates of eosinophils, lymphocytes and histiocytes, in addition to granuloma formation. In contrast, only minimal inflammation was observed in 3/10 mice in the control group (p=0.036).ConclusionAbatacept may play a role in inducing granulomatous hepatitis with a sarcoidosis-like reaction. Additional data including transaminases, antinuclear antibodies (ANA), Antimitochondrial antibodies (AMA) and other auto antibodies should be tested.

Electroacupuncture at "Zusanli" (ST36) and "Chize" (LU5) of mother rats exposed to nicotine during pregnancy and lactation has a protective effect on development of lung function and morphology in neonatal rats

To compare the different effect of electroacupuncture (EA) at "Zusanli" (ST36) and "Chize" (LU5) of mother rats exposed to Nicotine during pregnancy and lactation on lung function and morphological changes in offspring rats, so as to explore the most effective acupoint for improving the development of lung in neonatal rats. A total of 24 female pregnancy SD rats were randomly divided into 4 groups: normal control, model, EA-ST36 and EA-LU5 (n＝6 rats in each group). Rats of the normal group were treated by subcutaneous injection of normal saline, and those of the other 3 groups treated by subcutaneous injection of nicotine (1 mg•kg－1•d－1) beginning from the 6th day to about the 21st day of pregnancy (childbirth day) for nicotine exposure during pregnancy and lactation. The daily EA treatment (2 Hz /15 Hz，1 mA) was applied to bilateral ST36 and LU5 for 20 min, beginning from the 6th day of pregnancy to the 21st day (childbirth day). The lung function of the offspring rats including the peak inspiratory flow (PIF), peak expiratory flow (PEF), lung resistance (RL), exhalation resistance (RE)and lung dynamic compliance (Cdyn) was detected by using a lung function analysis system. Histopathological changes (severity of alveolarization) of the offspring rats' lung tissue were observed under microscope after H．E. stain. Compared with the normal group, the PIF, RL and RE values were significantly increased (P<0.01), and PEF and Cdyn values significantly decreased in the model group (P<0.01). The alveolar diameter in the model group was evidently increased relevant to the normal group (P<0.01). Following the intervention, modeling induced increase of PIF, RL, RE and alveolar diameter and decrease of PEF and Cdyn values in the EA-ST36 group, and the increased PIF, RL and RE levels in the EA-LU5 group were obviously suppressed relevant to the model group (P<0.01, P<0.05). Additionally, modeling induced obvious congestion and edema of the alveolar wall, alveolar deformation, rupture and fusion, and reduction of the number of the pulmonary alveoli were evidently milder in both EA-ST36 and EA-LU5 groups. No significant differences were found between the EA-ST36 and EA-LU5 groups in the abovementioned 5 indexes of pulmonary function and alveolar diameter (P>0.05)．. EA of ST36 and LU5 of mother rats experiencing nicotine exposure during pregnancy and lactation can improve the lung function and morphological changes in neonatal rats, and the effect of ST36 is relatively better.

The Effect of Subcutaneous and Intracutaneous Injections of Sterile Water and Normal Saline on Pain Intensity in Nulliparous Women: A Randomized Controlled Trial.

Background:Normal vaginal delivery is considered a painful process and it is difficult to tolerate the pain. The goal of this study was to compare the effect of injection of sterile distilled water and normal saline on pain intensity in nulliparous women.Materials and Methods:This triple-blind clinical trial was conducted on 164 nulliparous women randomly selected from among those who were hospitalized in Motahari Hospital of Jahrom, Iran, from 1 May 2012 to 1 October 2013. Women with a gestational age of 37–42 weeks, dilatation of 4–6 cm, and delivery 180 min after the intervention were selected. The subjects were randomly allocated to four groups of intracutaneous and subcutaneous sterile water and normal saline injections. Pain severity was measured 5 min before the injection and every 30 min up to 3 h after the injection using a visual analog scale. The data were analyzed using Chi-square, Scheffe, and Spearman's correlation tests in SPSS software.Results:There was no significant difference among the four studied groups concerning gestational age and other demographic characteristics. Chi-square test showed lower pain intensity 120 min after the injection in group 4 (subcutaneous injection of normal saline) (F3 = 14.75, p < 0.001) and 150 min after the injection in group 3 (intracutaneous injection of normal saline) (F3 = 14.75, p < 0.001). Chi-square test showed that the duration of the second stage of labor was shorter in group 4 participants (subcutaneous injection of normal saline) (F3 = −12.23, p < 0.001).Conclusions:The study showed that subcutaneous and intracutaneous injection of normal saline reduced the intensity of pain during childbirth.

Histological Survey of the Effect of Granulocyte-colony-stimulating Factor(G-CSF) on Bacterial Translocation and Wound Healing in Burned Mice

Background: Burn wound is an important cause of morbidity and mortality worldwide. Improving the host's immune system and removing the infection can be effective in healing wounds caused by burns. Granulocyte-colony-stimulating factor (G-CSF) stimulates both the bone marrow to produce granulocytes and the function of neutrophil precursors. The aim of this study was to examine the effect of G-CSF on removing infection and healing wound. Materials and Methods: A burn model was used to induce burns in 18 adult Balb/c mice, and their wounds were infected by Acinetobacter baumannii strains. Burned mice were divided into two groups (control and G-CSF) and treated daily by subcutaneous injections of normal saline (0.1 mL) and G-CSF (10 I¼g/kg). The wound healing process was evaluated by the morphological and histological assessments. Results: In morphological assay, the mean size of the wounds in the 3rd and 7th days of the treatment was significantly lower in the G-CSF treated group compared to the control group. Some of the histological parameters were evaluated, including the level of inflammation, re-epithelialization, angiogenesis, collagen deposition, the amount of granulation tissue, and fibroblast maturation. The results showed that inflammation was reduced in the G-CSF-treated group, and re-epithelialization and collagen deposition were increased insignificantly compared to the normal saline-treated group. Furthermore, bacterial translocation was reduced significantly in the G-CSF-treated group. Conclusion: G-CSF enhances wound closure and helps in wound healing by improving the immune system. It has also an anti-inflammatory role and reduces bacterial translocation.

Acute Histopathologic Findings Related to Needle Puncture Trauma during Subcutaneous Injection in the Sprague-Dawley Rat Model.

It is important to detect injection site reactions during the nonclinical phases of drug development. However, differentiating between normal changes following needle trauma and changes due to the toxicity of injected drugs can be challenging. Therefore, we used the Sprague-Dawley rat model to evaluate the pathological findings expected following a single subcutaneous injection of normal saline. Rats were subcutaneously administered with normal saline, and the injection sites were examined microscopically. Inflammation was evident in most of the injection sites, mostly in minimal severity. Parakeratosis/epithelial crust was also seen in several sites, and necrosis was observed in a minority of the cases. These findings indicate that needle puncture trauma can present with some degree of inflammation and necrosis. Although limited to a specific time point and strain, this study shows that inflammation following subcutaneous injection can be attributed in part to the needle trauma and not necessarily to the drug itself.

Effect of parathyroid hormone on cardiac function in rats with cardiomyopathy.

The present study investigated the role of parathyroid hormone (PTH) in non-ischemic cardiomyopathy (CM) and its underlying mechanism. A total of 30 Sprague-Dawley male rats were randomly divided into a control group (n=6) and an experimental group (n=24). To induce CM in the rats of the experimental group, 2 mg/kg Adriamycin (ADR) was administered intraperitoneally with 5 equal injections every third day followed by 5 weekly injections resulting in a cumulative dose of 20 mg/kg. Following establishment of the model, rats in the experimental group were subdivided into a PTH-untreated CM group that received daily normal saline subcutaneous injections for 7 days and three treated CM groups that received daily subcutaneous injections of 5, 10, or 20 µg/kg of recombinant PTH for 7 days. Rats in the control group accordingly received intraperitoneal and subcutaneous injections of normal saline. Blood sample analysis revealed that B-type natriuretic peptide (BNP), troponin T, C-reactive protein (CRP), creatinine and phosphorus concentrations were increased in the PTH-untreated CM group compared with that in the control group, whereas PTH and calcium concentrations were decreased. Administration of PTH dose-dependently decreased BNP, CRP, creatinine and phosphorus levels, and increased PTH and calcium levels. Notably, there were significant differences in PTH, BNP, troponin T, CRP, creatinine, calcium, and phosphorus levels among the rats in the five groups (P<0.01). Cardiac ultrasonography results indicated that the left ventricular ejection fraction (LVEF) was significantly decreased in rats treated with ADR compared with the rats from the control group (P<0.01). However, the LVEF gradually recovered with elevated PTH treatment doses. The overall differences of LVEF and left ventricular end-systolic volume in the five experimental groups were statistically significant (P<0.01). Furthermore, there were dose-dependent increases in LV mass and left ventricular end-diastolic volume in PTH-treated rats; however, the differences between any two groups did not reach statistical significance (P>0.05). Immunohistochemical staining and western blot analysis using an anti-PTH polyclonal antibody was performed to evaluate the protein expression levels of PTH in myocardial tissues. The mRNA expression levels of PTH and BNP were measured using reverse transcription-quantitative polymerase chain reaction. The results demonstrated that the mRNA and protein expression levels of PTH in myocardial tissues were significantly decreased in ADR-treated rats compared with the levels in the control group rats. Injection of recombinant PTH significantly increased PTH expression and reduced BNP expression in dose-dependent manners (P<0.05). These findings demonstrated that PTH can improve cardiac function in rats with ADR-induced CM, suggesting a potential therapeutic application for PTH in non-ischemic CM.

Effects of naloxone on opioid receptors in spinal cord dorsal horn of neuropathic pain rat models

Objective To investigate the effects of naloxone on the levels of opioid receptors in spinal cord dorsal horn of neuropathic pain rat models, established with chronic constriction injury(CCI). Methods Male SD rats (200-250 g) were subjected to CCI to establish neuropathic pain models. The rats were divided into 4 groups, and respectively received sham operation and saline treatment (Sham-S group, n=8), CCI and saline treatment (CCI-S group, n=8), CCI and naloxone treatment (CCI-N group, n=24), and CCI and morphine treatment (CCI-M group, n=8). Drug treatments (subcutaneous injection of normal saline, 10 mg·kg-1·d-1 naloxone, or 10 mg·kg-1·d-1 morphine in hindpaw) started from 7th day after surgery for 8 consecutive days. Paw withdrawal mechanical threshold (PWMT) and paw withdrawalthermal latency(PWTL) respectively were measured with von Frey tests and thermal plantar tests before and after CCI. Spinal cord dorsal horn at L3-L5 levels from naloxone-treated CCI rats were collected 10, 12, and 14 days after surgery, and western blot assay was employed to measure the levels of three opioid receptor subtypes. Results CCI rats exhibited ipsilateral hindpaw hyperalgesia, showing reduced PWMT and shortened PWTL (P 0.05) . Conclusions Naloxone, an opioid receptor antagonist, showed antinociceptive effects, similar to morphine, an opioid receptor agonist, on CCI neuropathic pain. This effect probably is relevant to naloxone-induced elevation of DOR in spinal cord dorsal horn. Key words: Naloxone; Neuropathic pain; Opioid receptors

Effect of Electroacupuncture on Pain Reaction and Content of Proteinase-activated Receptors 2 in Dorsal Root Ganglion in Hyperalgesia Rats

To observe the effect of electroacupuncture (EA) on mechanical hyperalgesia threshold (MHTs) and thermal hyperalgesia threshold (THTs) and content of proteinase-activated receptors 2 (PAR 2) in dorsal root ganglia (DRG) in rats with inflammatory pain, so as to explore its peripheral mechanism underlying improvement of inflammatory pain. The present study contains two parts. 1) In the first part, 27 male SD rats were randomized into sham hyperalgesic priming (sham-HP) group and real hyperalgesic priming (HP) group (n＝5 in the sham-HP group and n＝6 in the HP group for the test of MHTs, n＝8 in the two groups for the test of THTs). The sham-HP model was established by subcutaneous injection of normal saline into the left plantar part of the hind-paw, and the HP model established by subcutaneous injection of 1% carragenan (the first injection) into the same left hind paw, followed by injection of PGE2 (100 ng/25 μL, the second injection) into the dorsum pedis of the same hind paw 7 days after the first injection. The ipsilateral paw withdrawal latencies (MHTs and THTs) were detected before and 5 h, 3 d and 6 d after the first injection, 0.5, 4 and 24 h after the second injection. 2) In the second part, 64 male SD rats were randomly divided into sham-HP, HP, sham-EA and EA groups (n＝16 in each group). The sham-HP and HP models were made in the same way as the first part. Both"Zusanli"(ST 36)and "Kunlun"(BL 60) were punctured with filiform needles in the sham-EA group and also stimulated with EA: 2 Hz/100 Hz, 0.5－1.5 mA (0.5 mA increase per 10 min) for 30 min in the EA group, 1 time/d for 7 d. Both ipsilateral MHTs and THTs were observed at the same time-points of the first part and the PAR 2 protein content in the L 4－L 6 DRGs was assayed by ELISA 24 h after the second injection. 1) In the first part of the study, compared with the sham-HP group, the MHTs at 5 h and 3 d, and THT at 5 h after the first injection, and MHTs, and THTs at 4 and 24 h after the se-cond injection were significantly decreased in the HP group (P<0.01, P<0.05). 2) In the second part of the study, compared with the HP group, the MHTs at 4 and 24 h after the second injection and the THTs at 3 d after the first injection, 4 and 24 h after the second injection were significantly up-regulated in the EA group (P<0.01, P<0.05). The content of PAR 2 in the DRGs (L 4－L 6) was significantly higher in the HP group than in the sham-HP group (P<0.05), but considerably lower in the EA group than in the HP group (P<0.05). EA can suppress hyperalgesia priming in inflammatory pain rats which may be related to its effect in down-regulating PAR 2 level in the lumbar DRGs.

Wound Healing and Anti-inflammatory Effects of Topical Hyaluronic Acid Injection in Surgical-Site Infection Caused by Staphylococcus aureus.

Surgical-site infection (SSI) is a common postoperative complication, primarily caused by Staphylococcus aureus. S aureus produces hyaluronidase which degrades hyaluronic acid (HA). HA prevents bacterial proliferation and has anti-inflammatory effects to promote wound healing. We evaluated the effect of HA injection with systemic antibiotics for prevention and treatment of SSIs caused by S aureus. An open wound was created on the dorsum of 40 rats. The wound bed was sutured with S aureus inoculated thread. The test group was injected with HA (HA group), and the control group received a subcutaneous injection of normal saline (NS group). All groups were then treated with intraperitoneal cefazolin injection. The sutures were removed 2 days after the procedure. Gross pathology, bacterial count, and wound histology were assessed at days 2, 4, 6, and 8 postprocedure. The HA group showed a significant reduction in the wound area compared with the control group on gross pathology (at days 8 postprocedure, 36.54% ± 6.12% vs 50.59% ± 5.50%, P < .001). The HA group showed significantly better wound healing than the control group on histological analysis, including assessment of abscess, neutrophilic infiltration, and necrosis (4.2 ± 1.2 vs 11.5 ± 2.1, P < .001). The HA group showed a lower bacterial count compared with the NS group, but the result was not significant statistically (at days 6 postprocedure, 5.11 ± 0.31 vs 5.91 ± 0.35 logCFU/mL, P = .706). In conclusion, immediate local injection of HA in wounds can reduce SSI occurrence and promote wound healing in an animal model.

Effects of tumor necrosis factor-α monoclonal antibody on nuclear factor-κB activation and inducible nitric oxide synthase expression in rats with silicotic fibrosis

Objective: To investigate the effects of tumor necrosis factor-α (TNF-α) monoclonal anti-body on nuclear factor-κB (NF-κB) activation and inducible nitric oxide synthase (iNOS) expression in rats with pulmonary fibrosis induced by silica dust. Methods: A total of 48 male Wistar rats were randomly divided into intervention group, silica dust exposure group, and control group, with 16 rats in each group. The rats in the intervention group were given intratracheal injection of 50 mg silicon dioxide dust once to establish a rat model and then treated with subcutaneously injected TNF-α monoclonal antibody 15 mg/kg for 5 consecutive days at 2-6 days after the establishment of the model. The rats in the silica dust exposure group were treated with the same method to establish the model and then given subcutaneous injection of the same volume of normal saline. The rats in the control group were given intratracheal and subcutaneous injection of normal saline. In both groups, 8 rats each were sacrificed at 7 and 14 days after the establishment of the model. Hematoxylin-eosin staining or Masson staining was used to observe morphological changes in lung tissue, ELISA was used to measure the serum level of TNF-α, IHC was used to measure the expression of NF-κBp65 in lung tissue, Western blot was used to measure the protein expression of I-κB in lung tissue, and RT-qPCR was used to measure the transcriptional level of iNOS mRNA in lung tissue. Results: Compared with the control group, the silica dust exposure group had significant increases in the lung inflammation score (3.375±1.061 and 2.500±0.535) , serum TNF-α level (86.405±20.494 and 77.064±11.829) , absorbance of cells with positive NF-κBp65 in lung tissue (0.297±0.05 and 0.287±0.039) , and mRNA expression of iNOS (12.906±0.590 and 12.600±0.517) at 7 and 14 days after dust exposure, a significant increase in pulmonary fibrosis score at 14 days (3.250±0.707) , and significant reductions in the protein expression of I-κB at 7 and 14 days (0.579±0.141 and 0.748±0.081) (P<0.05) . Compared with the silica dust exposure group, the intervention group had significant reductions in the lung inflammation score at 7 days (2.375±1.061) , pulmonary fibrosis score at 14 days (2.375±1.061) , serum level of TNF-α at 7 and 14 days (66.565±19.850 and 58.734±16.335) , absorbance of cells with positive NF-κBp65 in lung tissue at 7 and 14 days (0.248±0.028 and 0.238±0.027) , and mRNA expression of iNOS at 7 and 14 days (11.656±0.405 and 12.025±0.618) , as well as significant increases in the protein expression of I-κB at 7 and 14 days (0.802±0.165 and 0.888±0.144) (P<0.05) . Conclusion: TNF-α monoclonal antibody can inhibit the activation of the NF-κB signaling pathway and down-regulate the expression of iNOS, and thus exerts a certain protective effect on lung tissue in rats with pulmonary fibrosis induced by silica dust.