A 3D hydrogel loaded with exosomes derived from bone marrow stem cells promotes cartilage repair in rats by modulating immunological microenvironment / 南方医科大学学报

OBJECTIVE@#To assess the efficacy of GelMA hydrogel loaded with bone marrow stem cell-derived exosomes for repairing injured rat knee articular cartilage.@*METHODS@#The supernatant of cultured bone marrow stem cells was subjected to ultracentrifugation separate and extract the exosomes, which were characterized by transmission electron microscopy, particle size analysis and Western blotting of the surface markers. The changes in rheology and electron microscopic features of GelMA hydrogel were examined after loading the exosomes. We assessed exosome release from the hydrogel was detected by BCA protein detection method, and labeled the exosomes with PKH26 red fluorescent dye to observe their phagocytosis by RAW264.7 cells. The effects of the exosomes alone, unloaded hydrogel, and exosome-loaded hydrogel on the polarization of RAW264.7 cells were detected by q-PCR and immunofluorescence assay. We further tested the effect of the exosome-loaded hydrogel on cartilage repair in a Transwell co-culture cell model of RAW264.7 cells and chondrocytes in a rat model of knee cartilage injury using q-PCR and immunofluorescence assay and HE and Masson staining.@*RESULTS@#GelMA hydrogel loaded with exosomes significantly promoted M2-type polarization of RAW264.7 cells (P < 0.05). In the Transwell co-culture model, the exosome-loaded GelMA hydrogel significantly promoted the repair of injured chondrocytes by regulating RAW264.7 cell transformation from M1 to M2 (P < 0.05). HE and Masson staining showed that the exosome-loaded hydrogel obviously promoted cartilage repair in the rat models damage.@*CONCLUSION@#GelMA hydrogel loaded with bone marrow stem cell-derived exosomes can significantly promote the repair of cartilage damage in rats by improving the immune microenvironment.

Value of LncRNA PRNCR1 in peripheral blood mononuclear cells as a diagnostic marker for osteoporosis and its relationship with inflammatory factors in patients / 中华内分泌外科杂志

Objective:To investigate the value of LncRNA PRNCR1 in peripheral blood mononuclear cells (PBMC) as a diagnostic marker for osteoporosis and its relationship with inflammatory factors in patients.Methods:The expression of LncRNA PRNCR1 in PBMCs of healthy controls and OP patients was detected by qRT-PCR and its diagnostic value in OP was explored. Enzyme linked immunosorbent assay (ELISA) was used to detect the concentrations of IL-6 and TNF-α in PBMC, and the correlation between LncRNA PRNCR1 and IL-6 and TNF-α concentrations was analyzed. LncRNA PRNCR1 was overexpressed or knocked down and its effect on inflammatory factor concentration was verified. The relationship between the expression of LncRNA PRNCR1 and the treatment days, recurrence rate of patients was analyzed, and the possible mechanism of LncRNA PRNCR1 affecting the progression of OP was explored through bioinformatics.Results:Compared with healthy control group, the expression of LncRNA PRNCR1 in PBMCs of OP group was significantly enhanced and could be used as a marker for the diagnosis of OP ( t=8.42, P<0.001) (AUC=0.83, P<0.001) . In addition, the concentrations of IL-6 and TNF-α in PBMCs of OP group were significantly enhanced and positively correlated with the expression of LncRNA PRNCR1 ( r=0.25, P=0.016) ( r=0.21, P=0.037) . Overexpression of LncRNA PRNCR1 promoted the expression of inflammatory factors while knockdown of LncRNA PRNCR1 inhibited the expression of inflammatory factors. Patients with high expression of LncRNA PRNCR1 remained on treatment longer than those with low expression. The results of bioinformatics analysis showed that LncRNA PRNCR1 may affect OP progression by affecting stem cell differentiation pathway and estrogen signaling pathway. Conclusions:The expression of LncRNA PRNCR1 in PBMC of patients with OP is significantly enhanced. LncRNA PRNCR1 can be used as an important marker for the diagnosis of OP and affects the expression of inflammatory factors.

Therapeutic effect of oral vaccine based on glutamate decarboxylase 65 on streptozotocin-induced type 1 diabetic mice / 中国药科大学学报

@#To investigate the therapeutic effect of oral vaccine based on glutamate decarboxylase 65 (GAD65) on streptozotocin (STZ) -induced type 1 diabetic (T1D) mice, the mice model of T1D was established by intraperitoneal injection of low dose multiple STZ. CTB-GADIII encapsulated with calcium alginate (Ca-Alg-GADIII) was formulated using crosslinking technology with sodium alginate and calcium chloride, and was administered intragastric to T1D mice once a week for 5 consecutive weeks.Blood glucose and body weight of the mice were recorded weekly, and pharmacodynamics against T1D of Ca-Alg-GADIII were investigated by glucose tolerance assay (OGTT) and pancreatic histopathological analysis. The levels of glutamic acid decarboxylase antibody (GADA), and insulin autoantibody (IAA) and related cytokines (IL-4, IFN-γ, TGF-β1) in serum were detected by ELISA, and the CD4 + T cell subsets were detected by flow cytometry. The immunological mechanism of oral vaccine against T1D was preliminarily discussed. The results showed that the disease-related indicators improved in immunized mice: fasting blood glucose improved, glucose tolerance and insulin secretion increased, pancreatic injury decreased, autoantibodies like GADA and IAA titers significantly decreased, and CD4 + T cell immune balance in mesenteric lymph node (MLN) and pancreatic lymph node (PLN) improved to some extent. The results suggest that oral vaccine Ca-Alg-GADIII has some therapeutic effect on STZ-induced T1D mice.

Feasibility of repairing articular cartilage defects with particulated juvenile cartilage allograft / 中国组织工程研究

BACKGROUND: Particulated juvenile cartilage allograft is simple and easy to obtain, and relevant clinical studies are underway in the USA. However, how the transplanted juvenile cartilage fragments exert biological effects through biochemical mechanisms and genetic signal transduction is still unclear. There is as yet no report on this technology in China. OBJECTIVE: To explore the feasibility of articular cartilage defects repaired with particulated juvenile cartilage allograft. METHODS: The cartilage fragments were obtained from juvenile Pitman-Moore strains (provided by the Laboratory Animal Center of the Army Medical University in China) and cultured in vitro. Brdu immunofluorescence assay was performed at 1, 3, and 7 days of culture. The particulated juvenile cartilage allograft/fibrin gel composites were subcutaneously transplanted into the SCID rats (provided by the Laboratory Animal Center of the Army Medical University). The specimens were taken for hematoxylin-eosin staining, safranin O staining and immunohistochemistry after 1 month. Cartilage defects of 8 mm in diameter were made in the knee joint of 10 adult Pitman-Moore strains (Laboratory Animal Center of the Army Medical University), and were randomized into two groups, which were then transplanted with the particulated juvenile cartilage allograft/fibrin gel composites (experimental group) or nothing (control group). The specimens were taken for hematoxylin-eosin staining, safranin O fast green staining, toluidine blue and immunohistochemistry at 3 months after transplantation. RESULTS AND CONCLUSION: Little Brdu incorporation was detected in juvenile cartilage fragments at 1 day of culture, some Brdu incorporation was defected at 3 days of culture. At 7 days of culture, a progressive increase in the Brdu signal was detected in chondrocytes within the cultured cartilage fragments, which seemed to localize along the tissue edge. At 1 month after subcutaneous transplantation, the particulated juvenile cartilage allograft still survived and were surrounded by few proliferative chondrocytes. There was no obvious tissue repair in the control group at 3 months after transplantation. In the experimental group, there was obvious tissue repair, the color of the newly formed tissues was similar to the normal cartilage tissue, which integrated well with the surrounding normal cartilage tissue, and the cells distributed evenly. These results imply that particulated juvenile cartilage allograft can achieve good results in repairing articular cartilage defects.

Application of microbial vaccines in type 1 diabetes mellitus / 中国药科大学学报

@#Type 1 diabetes mellitus(T1DM)is an autoimmune disease characterized by the destruction of β cells and insulin absolutely deficiency upon antigen-specific T lymphocyte attack. The patients with T1DM need insulin therapy for the whole life. Although the mechanism of T1DM is still unclear now, environmental factors play a critical role in the T1DM development because the incidence of infectious diseases is opposite to that of allergies and autoimmune diseases. So far, a variety of bacteria, parasites, viruses and their composition have been proved functional in the prevention of T1DM in animal models. Therefore, microbial vaccine, if administrated early, can induce immune tolerance and prevent the destruction of pancreatic islet β cells, which is expected to be safe and effective novel approach in treating T1DM. This paper summarizes several types of T1DM microbial vaccine and their potential mechanisms including inactivated vaccine, attenuated vaccine, subunit vaccine, DNA vaccine and living-vector vaccine.

Enhanced tenogenic differentiation by Scleraxis overexpression in human amniotic mesenchymal stem cells / 中国组织工程研究

BACKGROUND: Human amniotic mesenchymal stem cells (hAMSCs) are adult stem cells with multipotential differentiation, which can be induced to differentiate into bone, cartilage and other connective tissues. Meanwhile, as a highly specific marker of tenocytes, Scleraxis is involved in aggregation and differentiation of tendon progenitor cells as well as the formation of tendon extracellular matrix. OBJECTIVE: To investigate whether hAMSCs have the ability of differentiation into tenocytes by ectopic expression of Scleraxis. METHODS: Agreed by puerpera, the amniotic membrane from the full-term placenta was separated, and hAMSCs were isolated by a two-step enzyme digestion, observed under inverted phase contrast microscope, and identified by flow cytometry. Passage 3 cells were induced via plasmid-mediated Scleraxis overexpression in overexpression group. Untransfected cells cultured in normal medium served as blank control group, and those with empty plasmid transfection were defined as empty plasmid group. Cell proliferation was tested in each group using cell counting kit-8 within 7 days of culture. Real-time quantitative PCR and western blot were used to assess the tenogenic differentiation of hAMSCs in each group at 3 and 7 days of culture. RESULTS AND CONCLUSION: Findings from the cell counting kit-8 indicated that the cell viability had no significant differences among the groups within 7 days of culture (P > 0.05). Western blot results showed the protein expression of Scleraxis in the treatment group was significantly higher than that in the other two groups (P < 0.05). Real-time PCR results showed, at 3 days of culture, the expression of collagen type I, collagen type III, Fibronectin and Tenascin-C in the overexpression group was significantly higher than that in the empty plasmid group (P < 0.05), but the expression of Tenomodulin had no difference (P > 0.05); at 7 days of culture, the expressions of collagen type I, collagen type III, Fibronectin, Tenascin-C and Tenomodulin in the overexpression group were significantly higher than those in the empty plasmid group (P < 0.05). In summary, hAMSCs can be differentiated into tenocytes by ectopic expression of Scleraxis.

Association between chronic kidney disease and dilated Virchow-Robin space / 中华老年心脑血管病杂志

Objective To study the association between chronic kidney disease (CKD) and dilated Virchow-Robin space (dVRS) in acute lacunar ischemic stroke patients.Methods A total of 1078 acute lacunar ischemic stroke patients admitted to our hospital were divided into mild dVRS group 1 (n=737) and moderate-severe dVRS group 1 (n=341) according to the severity of their dVRS in basal ganglia (BG),and into mild dVRS group 2 (n=789) and moderate-severe dVRS group 2 (n =289) according to the severity of their dVRS in centrum semiovale (CSO).Their kidney function was assessed according to the estimated glomerular filtration rate (eGFR).CKD was classified into stage 1,stage 2,stage 3a and stage 3b.The association between renal function and dVRS was analyzed by multivariate logistic regression analysis.Results The age was older,the number of females was greater,the incidence of hypertension and CKD was higher,the proportion of smoking was lower in moderate-severe dVRS group 1 than in mild dVRS group 1 (P＜0.05).The incidence of hypertension was higher in moderate-severe dVRS group 2 than mild dVRS group 2 (P＜0.05).Multivariate logistic regression analysis showed that age,hypertension,stage 2 CKD,stage 3a CKD and stage 3b CKD were the independent risk factors for severe dVRS in BG (P＜0.05,P＜0.01).Hypertension was an independent risk factor for severe dVRS in CSO (P=0.04).Conclusion CKD is an important risk factor for dVRS in BG.However,it is not associated with dVRS in CSO.This result highlights the different pathological mechanisms and risk factors for dVRS in BG and CSO.

Effect of water conservancy schistosomiasis control projects combined with molluscicide to control Oncomelania hupensis snails in rivers connecting with Yangtze River in Pukou District,Nanjing City / 中国血吸虫病防治杂志

Objective To evaluate the effect of the water conservancy schistosomiasis control projects combined with mollus-cicide to control Oncomelania hupensis snails in the rivers connecting with the Yangtze River. Methods The water conservancy schistosomiasis control projects of Zhujiashan River,Qili River and Gaowang River were chosen as the study objects in Pukou District,Nanjing City. The data review method and field investigation were used to evaluate the effect of the water conservancy schistosomiasis control projects combined with molluscicide to control O. hupensis snails. Results After the projects of the wa-ter level control and concrete slope protection and mollusciciding were implemented,the snails in the project river sections were completely eliminated. The snail diffusion did not happen in the inland irrigation area too. In the outside of the river beach , though the snails still existed,the snail densities plunged below 1.0 snail per 1.0 m2. Conclusion The comprehensive mea-sures of the combination of water level control,concrete slope protection and mollusciciding can effectively control and eliminate the snails,and prevent the snails from spreading.

Articular cartilage defect repair with particulated juvenile cartilage allograft: existing problems and prospects / 中国组织工程研究

BACKGROUND: Particulated juvenile cartilage allograft is simple and easy to obtain, and chondrocytes can migrate and proliferate as confirmed by in vitro culture.In the Unite States,this technique has been used in the repair of cartilage defects in the hip, knee, ankle, and elbow joints. OBJECTIVE: To review the present situation, application, and value of particulated juvenile cartilage allograft transplantation for articular cartilage repair. METHODS: A computer-based search of CNKI, PubMed, and Elsevier was performed for retrieving articles concerning particulated juvenile cartilage allograft transplantation for articular cartilage repair published from October 1983 to June 2017. The keywords were "allogeneic juvenile cartilage particles; cartilage tissue engineering; articular cartilage defects;repair" in Chinese and English, respectively. After initial screening of titles and abstracts and exclusion of irrelevant articles, 48 eligible articles were included in final analysis. RESULTS AND CONCLUSION: (1) Although a variety of treatments for cartilage repair have achieved good clinical outcomes in short-term follow-up, improving the motor function of patients and relieving pain, patients eventually develop progressive degeneration of the articular cartilage and suffer from osteoarthritis. (2) Chondrocytes from allogeneic juvenile cartilage particles have stronger ability of proliferating and repairing cartilage defects in vitro than mature chondrocytes,and have low antigenicity,which cannot cause a strong rejection after in vivo transplantation.What's more, particulated juvenile cartilage allograft transplantation can be performed as one-stage surgery if cartilage defects are confirmed under arthroscopy. (3) Particulated juvenile cartilage allograft transplantation has achieved good outcomes in basic and clinical studies in the United States. Its potential superiority has gradually been accepted by doctors and patients. (4) There are also risks for being contaminated and spreading diseases during the preparation of particulated juvenile cartilage allograft. This technology has been widely used in the United States, but there are rare data concerning its follow-up studies. Therefore, an investigation on its long-term follow-up is indispensable for the objective assessment of its long-term efficacy, with a view to the extensive promotion of this technology in the clinical practice.

Analysis of posterior lumbar interbody fusion (PLIF) in treating lumbar degenerative disease in elderly patients / 中国骨伤

<p><b>OBJECTIVE</b>To explore the clinical effects of PLIF surgery for elderly patients with lumbar degenerative disease.</p><p><b>METHODS</b>From March 2010 to May 2013, 28 patients with lumbar degenerative disease, aged more than 80 years were treated with PLIF surgery. There were 10 males and 18 females, aged from 80 to 93 years old with an average of (85.44±3.66) years. Course of disease was from 3 to 20 years. The operation time, intra-operative blood loss, operation complications were recorded and JOA scores and Macnab criteria were used to evaluate the clinical outcomes.</p><p><b>RESULTS</b>All patients were followed up from 12 to 40 months with an average of 26.5 months. The average operation time was (150.00±26.42) min and the average intra-operative blood loss was (373.33±99.88) ml. The pre-operative JOA score was 12.30±2.43, and the corresponding postoperative JOA score at the final follow-up was 24.81±2.09 which was much higher than the preoperative one (P<0.01). According to the modified Macnab criteria to evaluate at the final follow-up, 16 patients got an excellent result, 10 good, 2 fair. In the weeks postoperatively, injuries of nerve root happened in 3 cases, superficial wound infection with delayed healing in 3 cases, and tear of the dural sac accompanied with cerebrospinal fluid leakage in 1 case. After long term follow-up, adjacent segment degeneration and the corresponding spinal canal stenosis occurred in 1 case at 34 months after operation. All cases got successful fusion without any displacement of internal fixation and pseudoarthrosis formation.</p><p><b>CONCLUSION</b>With proper cases, fully preoperative preparation, perfect intra-operative manipulation and active treatment after operation, even advanced ages older than 80 years with lumbar degenerative disease could get satisfactory outcomes after PLIF surgery.</p>

Change s of haemagglutination inhibitionan tibody level within one month after in fluenza A (H1N 1) vaccination / 中华微生物学和免疫学杂志

Objective To find the changes of haemagglutination inhibition ( HI ) antibody level against A/California/07/2009 (H1N1) within one month after pandemic A/H1N1 influenza vaccine (A/H1N1InfV) vaccination, and to provide data for drawing up immunization protocols against novel influenza . Methods The HI antibodies against A/California/07/2009 (H1N1) in sera from the inoculated subjects were tested by HI test .The geometric mean titer ( GMT) , geometric mean increase ( GMI) , seroconversion (SC) rate, seroprotection (SP) rate of HI antibodies were compared among the sera collected on day 3, 7, 14, 30 post vaccination .Results 961 participants were injected with A/H1N1InfV.In subjects aged 3 to 11 years, the antibody level peaked on day 14 post vaccination, but neither on day 14 nor on day 30, the lower bound of the two -sided 95%CI for the SP rate could fulfill the criteria of the FDA for influenza vac-cine.In subjects aged 12 to 60 years, the antibody level peaked on day 14 post vaccination and the SC rate , SP rate and GMI fulfilled the criteria of the European Medicines Agency ( EMEA) and the FDA for influenza vaccine. In subjects aged more than 60 years, the antibody level peaked on day 30 post vaccination , and the SC rate, SP rate and GMI on day 30 fulfilled the criteria of the EMEA and the FDA .Conclusion One dose A/H1N1InfV vaccination was able to induce enough protection on day 14 for subjects aged 12 to 60 years, on day 30 for subjects aged more than 60 years;however , for subjects aged 3 to 11 years who were antibody-negative at baseline , the lower bound of the two-sided 95%CI for the SP rate on day 14 and day 30 couldn′t fulfill the criteria of the FDA for influenza vaccine .

Treatment of degenerative lumbar spondylolisthesis through posterolateral fusion and fixation with pedicle screws / 中国骨伤

<p><b>OBJECTIVE</b>To explore internal fixation whether can improve the clinical outcomes of decompression and posterolateral fusion in patients with degenerative lumbar spondylolisthesis.</p><p><b>METHODS</b>From December 1998 to December 2005, 49 patients who had degenerative lumbar spondylolisthesis underwent decompression and posterolateral fusion without (group A, 21 cases) or with (group B, 28 cases) internal fixation (CD HORIZON M8 system). There were 12 males and 37 females with a mean age of 58.5 years (range, from 49 to 68 years). Among them,32 cases were grade I and 17 were grade II according to Meyerding grade system. All patients were followed up with an average of 58 months (range from 12 to 90 months). The pain of low back and leg (VAS scoring), spinal active function and neurologic function were evaluated according the assessment system of Yuan.</p><p><b>RESULTS</b>The VAS score of low back in group A and B were respectively 41.9 +/- 7.5 and 32.8 +/- 6.2 at follow-up; and VAS score of leg in group A and B were respectively 33.9 +/- 7.3 and 30.8 +/- 6.2. Spinal active function of patients, 15 cases obtained improvement, 6 cases aggravation or no improvement in group A; 25 cases obtained improvement, 3 cases aggravation or no improvement in group B. Neurologic function of patients, 15 cases obtained improvement, 6 cases aggravation or no improvement in group A; 26 cases obtained improvement, 2 cases aggravation or no improvement in group B. The group B was better than group A in the aspect of low back pain, spine active function and neurologic function (P < 0.05). There was no statistically significant difference in improvement of leg pain between two groups (P > 0.05).</p><p><b>CONCLUSION</b>Using internal fixation in decompression and posterolateral fusion for degenerative lumbar spondylolisthesis can improve low back pain and clinical function. Decompression is necessary for the surgical treatment of degenerative spondylolisthesis, which is major effect on the improvement of leg pain.</p>

Cell-wall-deficient bacteria: a major etiological factor for psoriasis? / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Psoriasis is a common inflammatory skin disease, yet knowledge of the factors that may induce, trigger, or exacerbate psoriasis is not fully delineated. Recent advances have improved our understanding of the link between psoriasis and cell-wall-deficient bacteria (CWDB) infections. In the present study we assessed the prevalence of CWDB infection in patients with psoriasis.</p><p><b>METHODS</b>The carriage rate of CWDB in the tonsil or pharynx of psoriasis patients, chronic tonsillitis patients and controls were investigated using hypertonic medium. Psoriasis patients with CWDB were randomly assigned to two groups and respectively treated with antibiotics or systemic therapy without antibiotic. Human peripheral blood mononuclear cells (PBMC) from psoriasis patients, chronic tonsillitis patients and control subjects were stimulated with bacteria antigens and extra-cellular levels of interferon-gamma (IFN-gamma) and interleukin (IL)-10 were measured in the supernatants using the ELISA technique, in vitro. Meanwhile, the proliferation ability of PBMC to respond to bacteria antigens was detected by MTT assay.</p><p><b>RESULTS</b>CWDB were isolated from 74.2% of psoriasis patients, 23.5% of chronic tonsillitis patients and only 6.3% of controls. Antibiotic therapy was appropriate for approximately 80% of psoriasis patients with CWDB infection, and in only 8.9% psoriasis patients CWDB infection was detected after antibiotic therapy. Meanwhile, our study showed that CWDB and wide-type bacteria did remarkably enhance the production of IFN-gamma, in vitro, and PBMC proliferation.</p><p><b>CONCLUSION</b>CWDB infection may be a virtual triggering factor in psoriasis by regulating T-cell activation.</p>

Molecular Epidemiology and Sequencing of the G-L Intergenic Region of Rabies Viruses Isolated in China / 中国病毒学

A group of 25 rabies viruses (RABVs),recovered from 24 dogs and one human case,were collected from various areas in China between 2004 and 2006.Genetic and phylogenetic analyses of the G-L intergenic region were carried out in 25 street RABV isolates and CTN vaccine strains of 7 generations.The study was based on the comparison of a 519 bp nucleotide sequence,encompassing the G-L intergenic region.The nucleotide sequence homologies of Chinese street strains were from 95.5% to 100%.The phylogenetic analysis showed that all Chinese isolates clearly supported the placement of all Chinese viruses in Lyssavirus genotype 1 and they were distributed according to their geographical origins.All of the Chinese strains were closely related but they could still be divided into two groups:group of street strains and group of CTN strains.This study presents details about the molecular epidemiology of rabies viruses based on the sequences of the G-L Intergenic region.

Detection and Genetic Characterization of Rabies Virus from Human Patients / 中国病毒学

Saliva and blood were collected from two patients who had not received post exposure prophylaxis in the cities of Wenzhou and Xinning respectively. Both patients were confirmed as positive for rabies by detection of rabies virus specific nucleoprotein antibodies in the sera by Western Blot. However, rabies virus specific RNA was only identified in the saliva collected from the patient in Wenzhou. Furthermore, the isolate Zhejiang Wz0 (H) was obtained by inoculating one-day-old suckling mice. Both nucleoprotein (N) and glycoprotein (G) genes from the isolate were amplified by RT-PCR and sequenced. Phylogenetic analysis indicated that the isolate belonged to classic rabies virus, and shared a higher homology with the street viruses from dogs in the main endemic areas in China and the street virus from dogs in Indonesia than with other known strains. Further comparison of the deduced amino acid sequences between the isolate and the vaccine strains used in China showed that the virus had a higher level of homology with the vaccine strain CTN than with the other vaccine strains (3aG, PV, PM and ERA). In particular, amino acid residues substitutions located in antigenic site Ⅲ in the G protein, which could react with the neutralizing antibodies, were observed. These results suggested that the virus belonged to the classic rabies virus, and both N and G genes diverged from the current vaccine strains used in China at either the nucleotide or the amino acid level.

An epidemiological study of rabies virus in domestic dogs, cats and wildlife and the immunogenicity study for rabies vaccines derived from different cell cultured virus strains / 病毒学报

For epidemiological investigation of the rabies virus carrier rates of domestic dogs, cats and wild animals like rodent animals and bats,three kinds of regions where rabies had higher incidence (Hunan and Guizhou Provinces), lower incidence (Jiangsu Province, Wuhan City) and provisionally rabies-free (Shenyang City) were selected. Then the antigenic types, the genovariation of the isolaled viruses and the currently vaccine matching of the virus strains were analyzed. The results showed that in China the principal host of rabies is dog,the total virus carrier rate of the captured dogs was 2.56%, and the highest positive isolation rate was 20.0% in some monitoring site. However,there was no evidence about the rabies virus carrier rate in rodent animals,bats or other wild animals. The rabies vaccines which prepared from aG and CTN strains have already been produced successfully in China. The research showed that the nucleotide sequences of the newly isolated viruses were more similar with the glycoprotein gene of CTN strain. In order to evaluate the safety and the efficacy of the vaccines currently used, two groups (50 people each) were injected with vaccine of aG strain and CTN strain respectively in five surveillance points. The neutralizing antibody tested were 0.49 IU/mL-0.52 IU/mL and 6.7 IU/mL-7.53 IU/mL after the 7 and the 14 days of vaccine injection respectively. In addition, the rates of antibody positive seroconversion were 45.1%-47.9% and 100% respectively, and there was no moderate or severe adverse reactions observed. These data showed the vaccines have satisfactory effect on safety and protection.

An epidemiologle investigation of hantaviruses carried by rodent hosts in Hunan province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the hantavirus infection and their genotype in rodents in Hunan.</p><p><b>METHODS</b>Hantavirus antigens in the rat lungs from Hunan province were detected by immunofluorescence assay. Partial S and M segment in antigen-positive samples were amplified by RT-PCR, and then sequenced. The phyologenetic trees were constructed for the analysis of genetic characters of hantavirus.</p><p><b>RESULTS</b>A total of 344 rats were trapped in the main epidemic area of Hunan province, and hantavirus antigens were found in 6 of the 344 rats( 1.74% ).The phylogenetic trees constructed by partial S segment( nt 620-990) or partial G2 segment (nt 2001- 2301) showed that the hantaviruses carried by Rattus norvegicus, R . flabipectus and R. rattoides from Xiangxiang district were genetic subtype SEOV4. The virus carried by R. norvegicus in Ningyuan district was phylogenetically different from the known SEOV. The hantavirus carried by Mus musculus from Shimen district was genetic subtype HTNV4.</p><p><b>CONCLUSION</b>The hantaviruses in the main epidemic areas in Hunan province mainly belonged to SEOV, and R. flabipectus and R. rattoides carried the same genotype of SEOV as R. norvegicus.</p>

Molecule character of rabies virus in 3 counties in Hunan China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the rabies molecular biology features in animals between high incidence area of rabies and no rabies cases area in Hunan.</p><p><b>METHODS</b>detect saliva of dogs and brains of dogs and cats by direct immunofluorescence assay, review positive samples by RT-PCR, sequencing extract RNA virus for genetic analysis.</p><p><b>RESULTS</b>12 were detected rabies virus antigen and positive nucleoside acid in 82 dogs from Wugang city also 1 in 17 from Dongkou county; the positive rate: Wugang 14.63 percent, Dongkou 5.88 percent. No rabies virus was detected in 67 samples of dog brains from Fenghuang County. Also none in 28 samples of cat brains. Amplificating N gene of rabies virus from positive samples of dog brain's tissue (No Wg13, Dk13) by RT-PCR, it shows that homology of nucleoside acid between two strain of virus is 99.4 percent; also 99.1 percent of amino acid. The homology of nucleoside acid (amonio acid) among Wg13 stain and Chinese strain CTN and aG strain is 89.4 percent (98.2 percent) and 86.1 percent (95.1 percent); The homology of nucleoside acid (amonio acid) among Dk13 stain Chinese strain CTN and aG strain is 89.1 percent (98.0 percent), 86.1 percent (94.9 percent).Compare with isolated rabies virus from abroad, the homology between two strains and Indonesia is 92.8 percent and 93.2 percent, the most similar of them. The strains isolated from other countries including Japan, Sri Lanka and India are relatively lower; The sequence of gene Wg13 and Dk13 were taken replacement of amino acid.</p><p><b>CONCLUSION</b>Two strains are belong to type I rabies virus, comparing its N gene with current using vaccine strains, both are in same group, and homology are relatively higher.</p>

Expression of VEGF-C and angiogenesis, and lymphangiogenesis in papillary thyroid carcinoma / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationship between the expression of vascular endothelial growth factor C (VEGF-C) and angiogenesis and lymphangiogenesis in papillary thyroid carcinoma (PTC).@*METHODS@#Seventy-two PTC cases were divided into 3 groups according to the level of invasion: papillary microcarcinoma group (PMC group), intrathyroid carcinoma group (IPC group), and extrathyroid carcinoma group (EPC group). They were again divided into 2 groups according to lymph node metastasis: lymph node metastasis group and lymph node no-metastasis group. The expressions of VEGF-C, CD105 and vascular endothelial growth factor receptor-3 (VEGFR-3) were detected by SP method of immunohistochemical staining. The expression of VEGF-C was analyzed quantitatively by image analysis system, and the PI of VEGF-C (VEGF-C-PI), the number of MVD (microvessel density), and LVD (lymphaticvessel density) were obtained.@*RESULTS@#The VEGF-C-PI of lymph node metastasis group (23.15 +/- 3.75) was higher than that of lymph node non-metastasis group (14.54 +/- 2.93) (P <0.01). MVD was 35.25 +/- 2.06 in the PMC group, 41.75 +/- 5.46 in the IPC group, and 52.58 +/- 4.16 in the EPC group, which showed the elevatory tendency with the increase of invasion (P < 00.5). LVD was 6.00 +/- 0.81 in the PMC group, 13.80 +/- 1.81 in the IPC group, and 19.17 +/- 2.96 in the EPC group, which again showed the elevatory tendency with the increase of invasion (P <0.05). The LVD of lymph node metastasis group (19.56 +/- 2.45) was significantly higher than that of lymph node non-metastasis group (12.48 +/- 2.84) (P < 0.05). VEGF-C was positively correlated with MVD and LVD (r = 0.743, 0.90, P <0.01).@*CONCLUSION@#The expressions of VEGF-C and LVD are related to lymph node metastasis of PTC. MVD and LVD are related to the invasion of PTC. VEGF-C may play an important role in the angiogenesis and lymphangiogenesis.

Analysis on the epidemiology of rabies in Hunan province in the past 10 years / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the possible factors causing the increase of rabies cases and to provide references for the development of related prevention and control strategy in Hunan.</p><p><b>METHODS</b>Data was collected and analyzed on epidemic situation of rabies in the past ten years, and studies were carried in some counties.</p><p><b>RESULTS</b>From 1994 to 1999, the annual cases sustained between 17 and 78. However, the number of cases has continued to increase since 2000. The annual reported cases in 2001 and 2002 were 311 and 313, and accounted for 34.8% and 30.0% of the total cases in the whole country respectively. The epidemic areas were mainly located in the south and middle parts of Hunan. Furthermore, the epidemic areas had been expanding. In 1997, human rabies cases were only reported in 7 cities but increased to 12 cities in 2004.</p><p><b>CONCLUSION</b>Factors as increasing number but low inoculating rate to dogs, and incorrect treatment on the wounds etc. might have served important roles for the recurrence of rabies. Hence, it is necessary to take comprehensive preventive measures to control and prevent the epidemics of rabies in Hunan.</p>