BACKGROUND:Abnormal stress is an important factor causing intervertebral disc degeneration. To establish an ideal in vivoanimal model of intervertebral disc degeneration is of great significance for in-depth study on the related pathogenesis. OBJECTIVE:To develop anin vivo rabbit intervertebral disc model and to investigate the relationship between continuous tensile load and intervertebral disc degeneration. METHODS:Twenty-five New Zealand white rabbits aged 6 months old were randomly divided into three groups: blank control (n=5), sham (n=10) and experimental (n=10) groups. The blank control group received no intervention, and the L4/5 segments were removed at the 1st day. The intervertebral disc assistor was used to fix the L4 and L5 vertebral bodies in the experimental and sham groups, the L4/5 segments in the experimental group were loaded 1 MPa axial tensile force, and the L4/5 segments in both two groups were then removed at 14 and 28 days. The changes of L4/5 intervetebral space height and surrounding bone substance were observed by X-ray examination, the morphological changes of the intervertebral disc were observed by hematoxylin-eosin staining, the cell survival was detected by nitro blue tetrazolium staining and mRNA expression levels of aggrecan, collagen type Ⅱ and SOX9 in the intervertebral disc tissues were assessed by RT-PCR at each time point. RESULTS AND CONCLUSION:The radiological manifestations, histological changes, cell survival and mRNA expression levels of aggrecan, collagen type Ⅱ and Sox9 showed no significant difference between the blank control and sham groups. Comparied with the blank control group, in the experimental group, the L4/5 intervertebral space was narrowed with time, the articular surface was coarse, and the upper and lower corpus vertebrae edge appeared to have lip-shaped hyperplasia; the intervertebral disc cells distributed irregularly; the nucleus pulposus was in dehydration and deflation, annulus fibrosus arranged irregularly, and the vacuoles in notochord cells tended to disappear; the expression levels of aggrecan, collagen type Ⅱ and SOX9 were markedly downregulated. These findings suggest that the in vivo rabbit model of intervertebral disc is successfully established, in which continuous mechanical tensile load is further proved to directly cause intervertebral disc degeneration.

Objective To study the role of IL-2 in regulating allograft rejection mediated by alloantigen-specific CD8~+ T cell.Methods T cell proliferation in vivo at a single cell level was examined using the CFSE dilution assay. IL-2 expression by activated CD4~+ versus CD8~+ T cells was determined by intracellular cytokine staining. The ability of alloantigen-specific CD8~+T cells in mediating allograft rejection was studied using the islet transplantation model.Results CD8~+ T cells divided vigorously in vivo in the allogeneic hosts regardless the presence or absence of CD4~+ T cells. CD4~+ T cells, but not CD8~+ T cells, were the primary source of IL-2 when both subsets were present. However, CD8~+ T cells could express high levels of IL-2 in the complete absence of CD4~+ T cells. In 2C TCR transgenic (Tg) mice in which the 2C TCR transgene was selectively expressed on the CD8~+ T cells that specifically recognized alloantigen (Ld) of Balb/c origin, islet allografts from Balb/c mice was promptly rejected by the 2CTg recipients with mean survival time of only 8 days. In contrast, in 2CTg mice with a genetic deletion of the IL-2 gene (2CTg-IL-2KO mice), the alloantigen specific CD8~+ T cells failed to mediate the islet allograft rejection and all the Balb/c islets survived for more than 50 days.Conclusions CD8~+ T cells appear to be very plastic in producing and utilizing IL-2. In the presence or absence of CD4~+ T cells, CD8~+ T cells can use CD4~+ derived or self derived IL-2 for proliferation and effector function respectively. In an alloantigen specific TCR transgenic model, the effector function of CD8~+ T cells is strictly IL-2 dependent. Thus, in situations where graft rejection is mediated solely by the CD8~+T cells, blocking IL-2/IL-2R pathway may be critically important in preventing transplant rejection.

OBJECTIVE To determine the distribution of bacterial flora in hospital infection and to provide laboratory(evidence) for controlling hospital infection and selecting rationally antibiotics in clinic practice.METHODS All(isolates) were identified by routine procedure.MRSA and ESBLs-producing rate of Escherichia coli and Klebsiella pneumoniae were(examined.) RESULTS Among all these clinical infectious specimens,there were 202 strains of Gram negative bacilli,(accounting) for 40.9%(202/495);166 strains of fungi,accounting for 33.5%;621 strains of Gram positive cocci,for 20.6%(102/495).Candida albicans,E.coli,Pseudomonas aerugionosa,C.tropicalis and C.glabrata took the first five bacteria in infection.Analysis of drug resistant bacteria suggested that the isolated rate of ESBLs-producing strains in Staphylococcus aureus be 47.6%,be CNS in MRCNS 78.1% and MRSA in SA be 42.3%.CONCLUSIONS Multidrug resistance and fungus infection are the main risk factors in our hospital.We must improve means of treatment on clinical work and use antibiotic rationally to reduce the infection rate.

Epidemics of hand, foot and mouth disease (HFMD) have mainly been caused by Coxsackievirus A16 (CVA16) and Enterovirus A 71 (EV-A71), which circulated alternatively or together in the affected area. CVA16 has caused numerous outbreaks and epidemics in multiple countries and geographical regions, and has become an important public health problem. Based on an analysis of the complete VP1 coding region, all CVA16 strains can be divided into genotypes A, B1, and B2. Furthermore, genotype B1 can be divided into subgenotypes B1a, B1b, and B1c. After 2000, no reports of genotype B2 virus strains have been reported. All of the CVA16 strains reported in mainland China have belonged to subgenotypes B1a and B1b. Most CVA16-associated infections cause only mild symptoms; however, some CVA16 infections can lead to severe complications and even death. Vaccination is considered to be the most effective method to control the transmission and infection rate of this virus. A number of research groups are studying various vaccine types, including inactivated vaccines, genetic engineering vaccines, and DNA vaccines, amongst others. In this review, an overview is provided of the research advances in molecular epidemiology and vaccines of CVA16.
Animals ; China ; Coxsackievirus Infections ; epidemiology ; immunology ; prevention & control ; virology ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Humans ; Molecular Epidemiology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

Objective:To investigate the effect of IL-35 on inflammatory response and T cell response in allergic rhinitis.Methods: 37 patients(observation group) with allergic rhinitis and 35 healthy volunteers(control group) after allergen detection of allergic rhinitisin in our hospital from Jan 2012 to Jan 2016 were selected as study subjects.The peripheral blood of observation group and control group were collected,and the serum levels of IL-35 were detected by ELISA.The animal model of allergic rhinitis in mice was established,the peripheral blood of mice was collected,and the serum level of IL-35 and IgE were detected by ELISA.The eosinophils that infiltrated in nasal mucosa were detected after tissue biopsy in mice.The mouse spleen cells were isolated and the ovalbumin antigen was added in the culture medium,IL-35 was or was not added into the culture medium,the ovalbumin specific T cell responses was detected.The cytokines IL-2,IL-4,IL-5,IL-10,IL-13,IL-17,IL-23,IL-27 and TNF-α in culture supernatant of ovalbumin specific T cells were detected by ELISA.The expression of IL-2,IL-4,IL-5,IL-10,IL-13,IL-17,IL-23,IL-27 and TNF-α in ovalbumin specific T cells were detected by Real-time PCR.The activation of JNK,Erk1/2 and p38 signal pathway in ovalbumin specific T cells were detected by Western blot.Results: The serum level of IL-35 in observation group was significantly lower than control group(P<0.05).The results showed that the number of eosinophils which infiltrated in AR mice nasal mucosa was significantly higher than normal mice(P<0.05),while the serum level of IL-35 in AR mice was significantly lower than normal mice(P<0.05).Ovalbumin specific T cell reactivity assay showed that IL-35 could significantly inhibit the T cell response.ELISA and Real-time PCR results showed that IL-35 could significantly down regulate the expression of IL-4,IL-5,IL-13,IL-17,IL-23 and TNF-α,and up regulate the expression of IL-2,IL-10 and IL-27.The Western blot results showed that IL-35 can inhibit the activation of JNK,Erk1/2 and p38 signal pathway of ovalbumin specific T in cells.Conclusion: IL-35 can regulate the expression of inflammatory cytokines in inflammatory response and inhibit T cell response,thus reducing allergic rhinitis,the mechanism may be through regulation of JNK,Erk1/2 and p38 signal pathway activation.

Objective To investigate the correlation between the formation of pigmented biliary calculus and biliary H.pylori infection.Methods Bile from 35 patients with pigmented biliary calculus and 10 healthy controls was cultured for aerobic,anaerobic and H.pylori.The expression of H.pylori- DNA in bile,bile duct mucosa and pigmented calculus were determined by PCR.The expression of H. pylori associated protein in bile duct mucosa was determined by Western-blot and Warthin-Starry staining.Results H.pylori culture was negative in all bile samples.In 35 patients with biliary pigmen- ted calculus,H.pylori was detected by PCR in the center of calculus,bile and bile duct mucosa of 14.29%,31.43% and 56.67% patients,respectively.Among H.pylori-DNA positive bile samples,7 contained anti-CagA antibodies,and 6 contained Vac A.in addition to Vacuolating cytotoxin(35000), glycoprotein(30000),Urase Band Urase A.Bacteria resembling H.pylori by Warthin-Starry stainning were found in 7 of 30(23.33%)bile duct mueosal samples from patients with biliary pigmented calculus. H.pylori-DNA and its associated protein were not detected in all bile and bile duct mucosae samples from the healthy controls.Conclusions The evidence of H.pylori-DNA and its associated protein in biliary system might indicate the role of H.pylori in the formation of biliary pigmented calculus.

OBJECTIVETo explore the effect of Ginkgo biloba extract (GBE) on the function of alveolar polymorphonuclear neutrophils (PMN) in severe acute pancreatitis (SAP) rats complicated with lung injury (LI).

METHODSForty-eight adult SD rats were randomly divided into three groups, i.e., the sham-operation group, the SAP group, and the GBE treatment group, 16 in each group. The SAP model was successfully induced by retrograde injection of 5% sodium taurocholate solution into the biliopancreatic duct. Rats in the sham-operation group only received flipping of the duodenum. Those in the GBE treatment group received GBE intervention based on SAP model. Equal volume of normal saline was given to rats in the sham-operation group and the SAP group. Rats were sacrificed at 6 and 12 h after operation respectively. The lung tissue was sampled to evaluate the LI score. The wet/dry ratio (W/D) of lung tissues was detected. The activity of myeloperoxidase (MPO) was measured. Alveolar PMN was harvested by bronchoalveolar lavage. The content of neutrophil elastase (NE) in bronchoalveolar lavage fluid (BALF) was measured by enzyme-linked immunoabsorbent assay (ELISA). The percentage of CD11b/CD18 double positive PMN was detected using flow cytometry. The expression of intercellular adhesion molecule-1 (ICAM-1) and NE protein in the lung tissue was detected by Western blot.

RESULTSCompared with the sham-operation group, significant pathologic lesion occurred in the lung tissue of rats in the SAP group; the pathologic LI score, lung tissue W/D ratio, MPO, and NE content in BALF significantly increased, the expression of ICAM-1 and NE in the lung tissue was obviously up-regulated, and the percentage of CD11b/CD18 double positive PMN significantly increased (P < 0.01). Compared with the SAP group, pathological lesion of the lung tissue was obviously attenuated, and the above indices were all significantly declined in the GBE treatment group (P < 0.01).

CONCLUSIONSExpression of ICAM-1 in the lung tissue and the percentage of D11b/ CD18 double positive PMN were up-regulated in SAP rats complicated with LI, resulting in the adherence of PMN to pulmonary vascular endothelial cells, and then activating PMN to release NE and aggravate LI. GBE could alleviate LI through down-regulating the expression ICAM-1 and CD11b/CD18, and hindering the adherence and activation of PMN to pulmonary vascular endothelial cells.

Animals ; Bronchoalveolar Lavage Fluid ; cytology ; Ginkgo biloba ; chemistry ; Intercellular Adhesion Molecule-1 ; metabolism ; Lung Injury ; drug therapy ; etiology ; metabolism ; Neutrophils ; metabolism ; Pancreatic Elastase ; metabolism ; Pancreatitis ; complications ; drug therapy ; metabolism ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

C.citratus has been usedin many countries with a long history.Traditionally,it is applied as a food seasoning in cooking.It is also used in tea beverage and folk medicine as well.Modern application of C.citratus is focused on the development of citronella oil,which can be used for food additives,disinfectants,cosmetics,drugs and etc.C.citratus is also a potential plant in landscaping.Its special lemony flavor contains chemical constituents,mainly including citral,myrcene,linalool,geraniol,nerol,citronellol,and etc.The modern research showed that C.citratus had the main effects of anti-microbial,anti-inflammation,analgesia,anti-oxidation,anti-tumor,anti-anxiety,anti-hypertension,antihyperglycemia,and etc.With further studies,some new pharmacological properties of C.citrates are going to be discovered gradually.It is worthy of further research and development to meet the needs of the health industry.

Objective To analyse the effects of serum estradiol levels during controlled ovarian hyperstimulation (COH) on the outcomes of in vitro fertilization and embryo transfer (IVF-ET). Methods The clinical data of 472 patients undergoing IVF-ET with GnRH analogues recombinant FSH long protocol were retrospectively analysed. The area under the curve (AUC) of estradiol (E2) level was calculated during COH, and patients were categorized into groups according to the percentile of AUC of E2(AUCE2) during COH. The general characteristics and parameters related to the outcomes of IVF-ET were compared among groups. Results The 10th percentile and 90th percentile of AUCE2 were 3 347.0 pmoL/L and 14 414.3 pmol/L, respectively. Four hundred and seventy-two patients were divided into lower reaction group (AUCE2 3 347.0 pmol/L, n=48), normal reaction group (14 414.3 pmol/L>AUCE2 > 3 347.0 pmol/L, n=376) and higher reaction group (AUCE2≥14 413.3 pmol/L, n=48). There was no significant difference in age, body mass index, baseline follicle stimulating hormone level, time of treatment with gonadotropin, endometrium thickness on day of transfer and embryos transferred(P>0.05). Compared with lower reaction group and normal reaction group, the number of oocytes per retrieval and number of embryos frozen were significantly larger(P<0.01) and the mild/severe ovarian hyperstimulation syndrome rate was significantly higher in higher reaction group(P<0.05). There was no significant difference in fertilization rate, cumulative embryo score, high-grade embryo rate, clinical pregnancy rate and implantation rate among groups (P> 0.05). Conclusion Sustained snpraphysiological serum E2 levels during the COH process do not adversely affect the quality of oocytes and embryos, clinical pregnancy rate and implantation rate to some extent in IVF-ET.

Objective: To analyze the epidemic trend of viral hepatitis in Nanjing from 1989 to 2019 and predict the incidence in 2020, so as to provide reference for the prevention and control of viral hepatitis. Methods: The incidence data of viral hepatitis in Nanjing from 1989 to 2019 was retrieved from Nanjng Center for Disease Control and Prevention and National Infectious Disease Reporting System. The epidemic trend was analyzed by estimating the annual percent change ( APC ) and the average annual percent change ( AAPC ). The seasonal incidence of different types of viral hepatitis was analyzed by seasonal index. The autoregressive integrated moving average model ( ARIMA ) was built to predict monthly incidence rate of viral hepatitis in 2020. Results: The annual incidence rate of viral hepatitis was 62.00/100 000 in Nanjing from 1989 to 2019, showing a downward trend ( AAPC=8.4%, P<0.05 ). From 1998 to 2019, the annual incidence rates of hepatitis A, B, C and E were 1.98/100 000, 14.31/100 000, 2.30/100 000 and 2.60/100 000. The incidence of hepatitis A and B showed downward trends ( AAPC=-11.81%, -6.02%, both P<0.05 ); the incidence trend of hepatitis C was not obvious ( P>0.05 ); the incidence of hepatitis E showed an increasing trend ( AAPC=4.82%, P<0.05 ). From 2015 to 2019, the third and fourth quarters were the epidemic seasons of hepatitis A, B and C, while the first and second quarters were the epidemic seasons of hepatitis E. The ARIMA model predicted that the monthly incidence rates of viral hepatitis in 2020 would range from 1.26/100 000 to 3.69/100 000, among which hepatitis B ranged from 1.21/100 000 to 2.58/100 000, hepatitis C from 0.20/100 000 to 0.48/100 000, hepatitis E from 0.09/100 000 to 0.25/100 000. Conclusions The incidence of viral hepatitis in Nanjing shows a downward trend. Among different types of hepatitis, hepatitis B has a higher incidence. All types of hepatitis have epidemic seasons. It is predicted that the monthly incidence rates of viral hepatitis will be 1.26/100 000 to 3.69/100 000 in 2020.