OBJECTIVE To explore the potential mechanism of the effect of Xuebijing injection （XBJ） on neurological function and survival of rats after cardiac arrest （CA）/cardiopulmonary resuscitation （CPR） based on the S-nitrosoglutathione reductase （GSNOR）/S-nitrosoglutathione （GSNO） pathway. METHODS The CA/CPR rat model was established by ventricular fibrillation. Using a sham operation group as control， high-throughput sequencing was employed to analyze and mine the differentially expressed genes （DEGs）. Enzyme-linked immunosorbent assay was used to determine the contents of GSNOR and GSNO in the hippocampus； the active components of XBJ were screened and subjected to molecular docking analysis with GSNOR. The rats successfully modeled using the same method were divided into model group （n＝30）， inhibitor （GSNOR inhibitor） group （n＝30）， XBJ group （n＝30） and XBJ+inhibitor group （n＝30）， and a sham operation group （n＝30） was set up. Neurological function was evaluated and survival status was recorded at 3 hours， 24 hours and 3 days after the first 89） drug intervention. The contents of GSNOR and GSNO in the hippocampus of rats were determined in each group at the 0191） above time points， and the relationship of the contents of GSNOR and GSNO with modified neurologic severity scale （mNSS） score was analyzed. RESULTS GSNOR coding gene was differentially expressed between the model group and the sham operation group. Compared with the sham operation group， GSNOR content increased significantly in the hippocampus of rats in model group， while GSNO content decreased significantly （P＜0.05）. The active components of XBJ， such as 4- methylenemiltirone and salviolone， could be bound to GSNOR protein， with the binding energy lower than －6 kcal/mol， mainly connected by hydrogen bonds. Animal experiments revealed that mNSS score and GSNOR levels in the hippocampus of rats in the model group were significantly higher than those in the sham operation group （P＜0.05）， while GSNO levels and survival rate were significantly lower than those in the sham operation group （P＜0.05）. The above indexes of rats were improved significantly in administration groups， the mNSS score in the XBJ group was significantly lower than that in the inhibitor group， the content changes of GSNOR and GSNO in the inhibitor group were more obvious than those in the XBJ group， and the various indicators in the XBJ+inhibitor group were significantly better than the XBJ group and the inhibitor group （P＜0.05）. GSNOR content was positively correlated with the mNSS score， and GSNO content was negatively correlated with the mNSS score （P＜0.05）. CONCLUSIONS XBJ can improve the neurological function of rats and enhance their survival rates after CA/CPR， the mechanism of which may be associated with the down-regulation of GSNOR and the up-regulation of GSNO.

AIM:To investigate the ameliorative effect of nociceptin/orphanin(N/OFQ)on anxiety-like be-havior in nicotine(NIC)withdrawal-induced rats and its regulatory mechanisms on the expression of neurotransmitters as-sociated with the hypothalamic-pituitary-adrenal(HPA)axis and inflammatory factors.METHODS:Thirty-two adult male Sprague-Dawley rats were randomly divided into 4 groups:normal control group,NIC withdrawal model group,low-dose N/OFQ treatment group,and high-dose N/OFQ treatment group,with 8 rats in each group.To establish NIC with-drawal model,the rats in the NIC model and N/OFQ treatment groups were subcutaneously injected with NIC(0.4 mg/kg),twice a day for 7 consecutive days followed by 3 days of withdrawal.During the withdrawal period,the rats in the low and high-dose N/OFQ treatment groups received intracerebroventricular injection of N/OFQ(1 nmol or 10 nmol)once per day for 3 consecutive days.Ten minutes after the third administration,all rats underwent open filed(OF)and elevated plus maze(EPM)tests to detect behavioral changes.The serum concentrations of corticotrophin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),corticosterone(CORT),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 were measured by ELISA.The mRNA expression levels of TNF-α,IL-1β and IL-6 in the central nu-cleus of the amygdala(CeA)of the brain were detected by RT-qPCR.Histological changes in neuron morphology in the CA1 region of the hippocampus were observed under a light microscope following hematoxylin and eosin(HE)staining.Norepinephrine(NE)levels in the CeA of the brain were determined by HPLC.The protein expression of tyrosine hydroxy-lase(TH)in the CeA of the brain was detected by Western blot.RESULTS:Compared with the NIC withdrawal model group,rats in the low and high-dose N/OFQ treatment groups showed significant increase in the distance and time spent in the central area of the open field(P<0.05 or P<0.01),as well as significant increase in the number of entries and the per-centage of time spent in the open arms of the EPM(P<0.05 or P<0.01).Furthermore,both low and high-dose N/OFQ treatment groups significantly inhibited serum concentrations of CRH,ACTH and CORT in NIC withdrawal rats(P<0.01).N/OFQ administration also significantly reduced the levels of inflammatory cytokines TNF-α,IL-1β,and IL-6 in the serum,as well as expression levels of TNF-α,IL-1β and IL-6 mRNA in the CeA(P<0.05 or P<0.01).The treatment with N/OFQ at both doses significantly alleviated neuronal damage in the CA1 region of the hippocampus and markedly re-duced thelevels of NE and TH protein expression in the CeA of NIC withdrawal rats(P<0.01).CONCLUSION:N/OFQ alleviates anxiety-like behavior in NIC withdrawal rats through mechanisms related to the regulation of HPA axis hormone levels and inflammatory factors.

Objective To investigate the inhibitory effect of Sauchinone(Sch)on renal tissue fibrosis and to explore its mechanism in unilateral ureteral obstruction(UUO)mice.Methods Forty male ICR mice were divided into Sham,UUO-Model,Sch Low-dose,Sch High-dose,and positive control(Val)groups.Mice in the sham group underwent isolation of the left renal ureter without ligation,and mice in the UUO-Model group underwent stripping of the left proximal ureter for double ligation.Starting on day 2 after surgery,the Sch Low-dose group received 10 mg/kg Sch,the Sch High-dose group received 30 mg/kg Sch,and the Val group received 100 mg/kg valsartan by gavage once daily for 4 weeks.At the end of drug administration,serum creatinine(SCr),blood urea nitrogen(BUN),and the inflammatory factors tumor necrosis factor(TNF)-α and interleukin(IL)-6 were measured.Superoxide dismutase(SOD)activity,and malondialdehyde(MDA)and reactive oxygen species(ROS)content were detected in renal tissues.TNF-α and IL-6 mRNA levels were detected in renal tissues by reverse transcription-quantitative polymerase chain reaction.Pathological changes and collagen deposition,as well as transforming growth factor(TGF)-β1,Smad3,and connective tissue growth factor(CTGF)protein expression in mouse renal tissues were detected by hematoxylin and eosin and Masson staining,and immunohistochemistry.Collagen Ⅰ,CTGF,Smad3,nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and nuclear factor(NF)-κB protein expression in renal tissues were detected by Western Blot.Results SCr,BUN,TNF-α,and IL-6 were all significantly reduced in Sch Low-dose and Sch High-dose mice compared with the UUO-Model group(P＜0.05 or P＜0.01),while SOD activity was significantly higher and MDA and ROS levels were significantly lower in renal tissues(P＜0.05 or P＜0.01),and TNF-α and IL-6 mRNA in renal tissues were significantly reduced(P＜0.05 or P＜0.01).Microscopic observation of swollen renal tubules with thylakoid hyperplasia and collagen fiber deposition was significantly improved.Immunohistochemistry showed a significant reduction in TGF-β1,Smad3,and CTGF protein expression in renal tissues,and Western Blot showed significantly reduced levels of Collagen Ⅰ,CTGF,Smad3,and NF-κB protein expression(P＜0.05 or P＜0.01),while Nrf2 and HO-1 protein expression levels were significantly elevated(P＜0.05 or P＜0.01).Conclusions The amelioration of renal tissue fibrosis by sauchinone may be mediated via anti-inflammatory and antioxidative stress effects that modulate the expression of pro-fibrotic proteins in the TGF-β1/Smad3 signaling pathway.

【Objective】 To determine the reference range of thromboelastogram(TEG) and establish a TEG feature for local population by measuring TEG parameters in healthy adults in Shenzhen comparing the difference between gender and age, and analyzing the reference data provided by reagent manufacturer. 【Methods】 A total of 916 healthy adults, aged between 19 to 59, who did their regular health checks in our hospital from September 2020 to August 2021 were selected. The TEG(from Lepu Medical Technology Co., Ltd.) was performed, and the clot reaction time(R), clot formation time(K), coagulation angle(α-Angle), maximum amplitude(MA), coagulation index(CI), fibrinolysis index LY30 and the estimated percent lysis (EPL) were analyzed. 【Results】 The reference ranges of TEG parameters, including R, K, α-Angle, MA, CI, LY30 and EPL, of 916 healthy adults from Shenzhen were 3.25~8.19 min, 0.66~3.18min, 47.70~76.56deg, 50.05~72.91mm, -4.3~3.4, 0~2.2% and 0~3%, respectively. The value of α-Angle, CI, K, LY30, MA and R didn’t all meet the given range provided by the manufacturer; some were exceeding and some inferior to. A total of 227 out of 916 individuals presented abnormal results, relative to the references, in at least one parameter, and 78 were diagnosed of abnormal coagulation based on the given reference range, with a specificity of 75.2%. 【Conclusion】 The reference range of TEG parameters of Shenzhen locals is significantly different from that provided by manufacturers. And it is imperative for local TEG laboratories to establish their own reference ranges according to age and gender groups based on local population characteristics.

Objective To explore the method and effect of applying just-in-time-teaching (JiTT) mode in the reform of the teaching method of biochemical comprehensive experiment (Isolation, Purification and Identification of γ-Globulin From serum). Methods A total of 100 undergraduate students in our 2016 clinical medicine were divided evenly into two groups: the control group and the experimental group. The control group used traditional teaching, and the experimental group used JiTT-based teaching. In the experimental group, an online environment for pre-classroom discussion and feedback was constructed by uploading learning resources such as short video and tutorials to the online teaching platform and the online learning exchange group 3-5 days before the class; the class session itself consisted of personal tests, face-to-face teaching by teachers, group discussion, finishing experimental report and answering questions etc., and was intended to evaluate the students' self-study. SPSS 22.0 was used to collect exam scores and questionnaire results, and the data between groups were compared using t test. Results The scores of individual test scores in the experimental group (9.44±0.59) were higher than those in the control group (8.77±0.41) (P=0.00), and the difference was statistically significant. In all the six questionnaire items pertaining to teaching effect , JiTT teaching method received higher scores than the traditional teaching method (P<0.05), and the difference was statistically significant. Conclusion JiTT can improve the quality of biochemistry experiment teaching. It is worthwhile to try it in the teaching of other basic specialized courses.

Objective To observe the biological characteristics of db/db mice, and to provide the basis for application of db/db mice in experimental research.Methods Spontaneous type 2 diabetes BKS.Cg-Dock7m +/+ Leprdb/JNju mice and wild type mice of the same age were used in this study.Their fasting blood glucose was determined at 8,12,16, 20 and 24 weeks of age, the body weight was recorded at 10, 12,16, 20 and 24 weeks of age, the levels of serum insulin, total cholesterol and triglyceride were measured, the organ weight and liver coefficient were determined, and the liver and pancreas were taken for pathological examination at 24 weeks of age.Results The db/db mice maintained a high level of fasting blood glucose and body weight.Levels of serum insulin, total cholesterol, and triglyceride were significantly higher than the wild type mice.Compared with wild type mice, liver weight and kidney weight were also significantly increased.Obvious pathological changes of liver and pancreas were observed in 24-week old db/db mice.Conclusions db/db mouse has obvious characteristics of type 2 diabetes, such as hyperglycemia, hyperlipidemia, and hyperinsulinemia, can maintain stable levels of high blood glucose,and is an ideal animal model for experimental study of type 2 diabetes mellitus.

TOF-MS has been more and more widely used in clinical and scientific research due to its advantages such as fast and easily operated.Diabetic nephropathy is the most common complications of diabetes.But clinical existing detection meth-ods to some extent have some hysteresis.The applications and developments of TOF-MS coupled with other proteomics technologies provided new ideas and methods for early diagnosis and prognosis of diabetic nephropathy.In this paper,the lat-est research results in nearest years about the application of TOF-MS to diabetic nephropathy will be reviewed.

Objective To investigate the relationship between Hepatitis B patients with different viral loads and immunoglob-ulin A,G,M and complement C3,C4.Methods Firstly,followed by real-time fluorescence quantitative PCR detection 210 cases of hepatitis B patients with HBV-DNA levels,according to 10n copies/ml different viral load detection results,it was divided into 102 ~108 copies/ml of the experimental groups.Then the experimental groups and control group were simulta-neously detected in immunoglobulin A,G,M and complement C3,C4.Analysed the correlation between HBV loads and im-munoglobulin A,G,M and complement C3,C4.Results When the viral loads of hepatitis B patients were 105 ~108 copies/ml,the testing results of IgA,IgG and IgM were both increasing (U =12.43,10.96,6.42,P <0.01),while C3 and C4 were both decreasing (U =8.37,6.0,P <0.01).When the viral loads of hepatitis B patients were 102 ~ 104 copies/ml,only IgA and IgM were increasing (U =2.36,2.04,P <0.05),the other testing results had no statistical significance.Between the test of 7 experimental groups compared with each other,only 104 group and 105 group had significantly changed (IgA and IgM were increasing,C4 was decreasing,U =2.39,2.46,2.09,P <0.05,IgG was increasing,U = 3.25,P <0.01),but between other low viral loads or high viral loads were not significantly differences.Conclusion The different viral loads of hepatitis B patients could cause the different changes of immunoglobulin A,G,M and complement C3,C4,especially in the 4 groups from 105 to 108 copies/ml.Followed by increasing in viral loads,there were immunoglobulin A,G,M increasing and comple-ment C3,C4 decreasing,and also serious impaction on the immune function of organism.There was a positive correlation be-tween viral loads in vivo and immune damages,correlation coefficient (γ =0.967,P <0.01).When the viral loads from 104 to 105 copies/ml,the testing results had changed significantly.It suggest that should control viral loads under 104 copies/ml in the hepatitis B antiviral treatment,so the effect of immune function damage will be the minimum.

BACKGROUNDWe previously demonstrated that the aqueous extract of the Schizandra chinensis fruit (AESC) ameliorated Cd-induced depletion of monoamine neurotransmitters in the brain through antioxidant activity. In the present study, we investigated the effect of AESC on anxiety-like behavior and the levels of norepinephrine and 3-methoxy-4-hydroxy-phenylglycol (a metabolite of norepinephrine) in different brain regions during ethanol withdrawal in rats.

METHODSMale Sprague-Dawley rats were treated with 3 g/kg of ethanol (20%, w/v) or saline by daily intraperitoneal injection for 28 days followed by three days of withdrawal. During withdrawal, rats were given AESC (100 mg × kg(-1)× d(-1) or 300 mg × kg(-1)× d(-1), P.O.) once a day for three days. Thirty minutes after the final dose of AESC, the anxiogenic response was evaluated using an elevated plus maze, and the plasma corticosterone levels were examined by radioimmunoassay. Meanwhile, the concentrations of norepinephrine and 3-methoxy-4-hydroxy-phenylglycol in the hypothalamic paraventricular nucleus and hippocampus were also measured by high performance liquid chromatography.

RESULTSRats undergoing ethanol withdrawal exhibited substantial anxiety-like behavior, which was characterized by both the decrease in time spent in the open arms of the elevated plus maze and the increased level of corticosterone secretion, which were greatly attenuated by doses of AESC in a dose-dependent manner. The high performance liquid chromatography analysis revealed that ethanol withdrawal significantly increased norepinephrine and 3-methoxy-4-hydroxy-phenylglycol levels in the hypothalamic paraventricular nucleus, while not significantly altering them in the hippocampus. Similar to the results from the elevated plus maze test, the AESC significantly inhibited the elevation of norepinephrine and its metabolite in the hypothalamic paraventricular nucleus in a dose-dependent manner.

CONCLUSIONSThese results suggest that AESC attenuates anxiety-like behavior induced by ethanol withdrawal through modulation of the hypothalamic norepinephrine system in the brain.

Animals ; Anxiety ; drug therapy ; etiology ; Behavior, Animal ; drug effects ; Ethanol ; adverse effects ; Fruit ; chemistry ; Male ; Plant Extracts ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Schisandra ; chemistry ; Substance Withdrawal Syndrome ; drug therapy

Objective:To evaluate the stability and bio-safety of warming moxibustion gel-unguentums( WMG) . Methods: With the commercially available package and at 40℃ ± 2℃, RH (75% ± 5%) for six months,the stability of WMG was inspected. Guinea pig skin sensitization test, rabbit skin irritation test and cell toxicity test were used to investigate the safety. Results:Compared to the results of 0 month, there was no difference in the sample character, containing paste percentage, adhesion, shape, weight difference and all theindices in TLC identification. Cell toxicity test showed that WMG had slight cytotoxicity. Guinea pig skin sensitization test and rabbit skin irritation test indicated that the grade of skin reaction was zero and the primary stimulus index of skin tissues was also zero. Conclusion:The quality of WMG is stable and the production preparation technology is feasible. Moreover, it is safe in transder-mal administration.