The symbiotic bacterium exists in the intestines of entomopathogenic nematodes and is a potential biological agent.Systematic classification of these bacteria is scarce in China.In this paper,seven strains of symbiotic bacteria from local entomopathogenic nematodes were identified by both observation of mor-phology,physiological,biochemical characteristics and sequence analysis of 16S rDNA fragments.

Objective To investigate the epidemiological characteristics of Brucella aggregation of disease outbreaks, and explore the causes and possible routes of transmission in Tongxiang City for proposing prevention and control measures.Methods A case investigation was done to the first case of bovine brucellosis, while search of new case was carried out in the population.Epidemiological survey was carried out in accordance with the case study form from Chinese Center for Disease Control and Prevention that developed the unified form, at the same time, survey of risk factors were carried out.According to the principle of informed consent, blood samples were collected, brucellosis rose bengal plate agglutination test (RBPT) and tube agglutination test (SAT) were used, and positive specimens were identified by Brucella culture.Brucellosis diagnosis was based on Brucellosis Diagnostic Criteria (WS 269-2007).Results The first case was a male Fifty-six years old wine salesman from the farmers market.Serum of the patient was (+) in RBPT test, and 1 : 400 (++) in SAT test.Cattle Brucella was check out by blood culture.Totally 66 human serum samples from the case search in key groups were tested, three people were detected seropositive but Brucella culture result was negative.Three cases were determined latent infection who had contacted beef sales staff.Conclusions The survey has shown that it is possible to get infected through contact with respiratory, skin, mucous membranes of Brucella infected beef or contaminated environment.Epidemic form should be concerned of non-professional personnel who are infected with different isoforms of brucellosis.The training of clinicians should be strengthened to improve their diagnosis and treatment technology.Health education should also be carried out to raise awareness of key population of disease prevention.

ObjectiveTo detect the difference of cytokines and antibodies productions by immunologic system from mice and rabbits vaccinated with the M.RCAg-1 chimeric protein,expressed in E.coli,formulation with different adjuvants,including Freund's adjuvant and three clinically acceptable adjuvants,namely,Al(OH)3,Montanide ISA720 and Montanide ISA51.MethodsSix weeks female BALB/c mice were vaccinated with recombinant protein formulated with different adjuvants through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual Epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; Affinity assay between protein and immune IgG of rabbits with the biosensor,and the growth of Plasmodiumfalciparum in vitro to evaluate by growth inhibition assay(GIA).ResultsDifferent formulation can induce different levels of antibody titers,the effection of ISA51 adjuvant was most closely with Freund's adjuvant,and can induce a higher specific antibody of 11 epitopes within proteins,can effectively stimulate cellular immune response based on the IFN-γ,to avidity Montanide ISA51 adjuvant immune antibodies and M.RCAg-1 protein affinity than the other two adjuvants;and Montanide ISA720 adjuvants and Al (OH)3 adjuvant group in mice can't induce a significant IFN-γresponse(P＞0.05).On avidity assay,the Montanide ISA51 formulation group was better than the other two adjuvants; and Montanide ISA720 and Al (OH)3 adjuvant formulation group can't induce a significant IFN-γresponse in mice(P＞0.05) ; the inhibition rates were 60% and 100% in 3D7 and Dd2 Plasmodium falciparum at a concentration of 2 mg/ml IgG by Montanide ISA51 formulated protein,and IgG of Al( OH)3 formulation could not effectively inhibit the in vitro growth of Plasmodium falciparum( 10% ),while IgG of Montanide ISA720 formulation could not inhibit growth of parasite in vitro.ConclusionBy comparing three clinically acceptable adjuvants and Freund's adjuvant in BALB/c mice and New Zealand rabbit,Montanide ISA51 adjuvants can be acceptable formulated M.RCAg-1 protein induced humoral and cellular immune responses,can be used as one of the candidate adjuvants.

ObjectiveTo explore the effects of vector fusion peptides on the conformation and immune reactivity of recombinant polyepitope antigens,M.RCAg-1.MethodsWe subcloned polyepitope artificial antigen gene,M.RCAg-1,into prokaryotic expression vectors,pDS-ex,that contain different fusion tags at the N-terminus or no any tag by different restriction enzyme cutting site.Three recombinant proteins expressed by these vectors,named P312-1,P312-2,and P312-3,were purified and purity is greater than 95％.Then BALB/c mice were vaccinated with the three proteins formulated with Freund's adjuvant through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; and the growth of Plasmodium falciparum in vitro to evaluate by growth inhibition assay(GIA),and analyze secondary and tertiary structures of recombinant proteins from different expression vectors by bioinformatics and circular dichroism technique.ResultsThe P312-1,P312-2 have almost the same amino acid sequence,and the three proteins have the same immunogenicity in animal models(P＞0.05),however,the different proteins elicited various T-cell responses,the rabbit antibody induced by these proteins showed diverse efficacy in malaria parasite growth inhibition assaysin vitro ( respectively,93.9％,14.7％,54.3％ ).The significant differences of secondary and tertiary structures were shown in recombinant proteins from different expression vectors,analyzed by bioinformatics and circular dichroism technique,which demonstrated the change of protein molecule spaces conformation,and the obviously change of some epitope locations.ConclusionThese results suggest that the expressed polyepitope artificial antigens originating from the different vector fusion peptides indeed affect the protein folding and,subsequently,the epitope exposure.Thus,these proteins are able to induce both distinct humoral and cellular immune responses in animal models,and they affect the efficacy of immune inhibition against the parasite,the enhancement or suppresses.

Adult ; China ; Emergency Medical Services ; organization & administration ; Female ; Health Knowledge, Attitudes, Practice ; Health Personnel ; Humans ; Male ; Middle Aged ; Poisoning ; prevention & control ; therapy ; Professional Competence ; Sampling Studies ; Surveys and Questionnaires

As the largest research-oriented specialty department in national traditional Chinese medicine hospitals, the Department of Critical Care Medicine in Guangdong Provincial Hospital of Chinese Medicine insists on the development mode combined with clinical medicine and scientific research. By taking clinical and basic researches for integrative medicine preventing and treating acute myocardial in-farction and sepsis as a breakthrough, authors explored key problems of Chinese medicine in improving the prognosis related diseases and patients' quality of life. In recent 3 years our department has successively become the principal unit of the national key specialties cooperative group of critical care medicine (awarded by State Administration of Traditional Chinese Medicine), the key clinical specialties (awarded by National Health and Family Planning Commission), and Guangzhou key laboratory construction unit, and achieved overall lap in clinical medical treatment, personnel training, scientific research, and social service.
Biomedical Research ; China ; Clinical Medicine ; Critical Care ; Hospital Departments ; organization & administration ; Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; Quality of Life

OBJECTIVETo explore the relation of body mass index (BMI) and waist circumference (WC) of the aged with clustering of other cardiovascular risk factors.

METHODSTotal 654 old people were checked up, and the systolic blood press (SBP), diastolic blood press (DBP), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and fasting plasma glucose (FPG) of them were investigated according to different BMI and WC, then the other cardiovascular risk factors analysis was carried out on the detection rates of SBP, DBP, TC, HDL-C, FPG level and high blood pressure, high TC, high TG. The clustering of the other cardiovascular risk factors was analyzed.

RESULTSThe rising of BMI and WC was regular cardiovascular risk factor in the aged. The average level of SBP, DBP, TC, TG, and FPG of the aged was obviously rising along with BMI and WC increasing, meanwhile the HDL-C was obviously decreased, and the detection rates of high blood pressure, high TC, TG and diabetes mellitus were obviously increased. When the BMI was 24 kg/m(2)-27.9 kg/m2, the detection rates of high blood pressure, high TC, TG and diabetes mellitus were 59.74%, 3.89%, 28.57%, 10.06% respectively, and their risks of suffering high blood pressure, high TC and TG were 1.65, 1.88, 1.85 times of those with normal BMI. When the BMI > or = 28.0 kg/m2, the detection rates of high blood pressure, high TC, TG and diabetes mellitus were 83.05%, 5.08%, 35.59%, 15.25% respectively, and their risks of suffering high blood pressure, high TC and TG were 5.44, 2.60, 2.98 times of those with normal BMI. When the man WC > or = 85 cm or women WC > or = 80 cm, the detection rates of high blood pressure, high TC, TG and diabetes mellitus were 66.15.%, 4.47%, 29.57%, 10.12%, and their risks of suffering high blood pressure, high TC and TG were 3.52, 6.51, 1.68 times of those with normal BMI. The rate of the aged with several cardiovascular risk factors was significantly increased for those with BMI > or = 24.0 kg/m2 or man WC > or = 85 cm or women WC > or = 80 cm.

CONCLUSIONThe rising of BMI and WC should be important factor for the clustering of cardiovascular risk factors in the aged.

Age Factors ; Aged ; Blood Pressure ; Body Mass Index ; Cardiovascular Diseases ; blood ; etiology ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Female ; Humans ; Male ; Middle Aged ; Obesity ; epidemiology ; Risk Factors ; Triglycerides ; blood ; Waist Circumference

Objective:To study the combined use of endoscopic balloon dilation with endoscopic biliary brushings in diagnosis of bile duct strictures.Methods:A prospective single center study was conducted at the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology. All patients with suspected malignant bile duct strictures shown on CT or MRI imaging from January 2018 to January 2020 were reviewed. All patients gave informed consent to the endascopic retrograde cholangiopancreatography procedures. Their clinical and follow-up data were analyzed. All patients underwent endoscopic balloon dilation of bile duct strictures. Before and after balloon dilation, biliary brush cytology was performed, and the results were used to classify the patients into the control group and the experimental group. Pathological examination of the brush cytology samples was carried out by a single chief pathologist. Presence of cancer cells or significant heterogeneous cells indicated a positive brush cytology test. Negative patients who still highly consider cholangiocarcinoma and agree to surgery and whose gross specimen is confirmed to be malignant after surgery should be considered as false negative by brush examination; it is difficult to judge that patients with cholangiocarcinoma have progress after 2 months of follow-up should be considered as false negative by brush examination. Any progression of disease indicated that the brush test was wrong and the test was again classified as false negative. Only when there was no progression of strictures was the possibility of a benign biliary stricture being considered. The advantage test (McNemar test) was used to analyze the difference between the two diagnostic methods.Results:Of 39 patients who were included in this study, there were 26 males and 13 females, with an age of (68.0 ± 5.2) years. Cholangiocarcinoma was diagnosed by histopathology, surgery or at 2 months follow-up in 35 patients. In the control group, 17 patients had a positive brush test (sensitivity rate was 48.6%, 17/35). In the experimental group, 26 patients had a positive brush test (sensitivity rate was 74.2%, 26/35). In addition, 2 patients in the control group had a positive brush test, while in the experimental group, a negative brush test. A total of 28 patients were positive in the two groups. The sensitivity rate of the brush test was 80.0% (28/35). There were significant differences between the two groups ( P<0.05). Conclusion:Endoscopic balloon dilation combined with endoscopic biliary brushings improved the sensitivity of pathological diagnosis of cholangiocarcinoma, and endoscopic biliary brushings before and after balloon dilation improved the sensitivity of diagnosis.

OBJECTIVE: To explore the correlation between postmortem interval (PMI) and five RNA markers of rat's skin--β-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S ribosomal RNA(18S rRNA), 5S ribosomal RNA (5S rRNA), and microRNA-203 (miR-203), at different temperatures. METHODS: Eighteen SD rats were randomly divided into three environmental temperature groups: 4 °C, 15 °C and 35 °C, respectively. Skin samples were taken at 11 time points from 0 h to 120 h post-mortem. The total RNA was extracted from the skin samples and the five RNA levels were detected by real-time fluorescent quantitative PCR. Proper internal reference was selected by geNorm software. Regression analysis of the RNA markers was conducted by GraphPad software. RESULTS: 5S rRNA and miR-203 were most suitable internal references. A good linear relationship between PMI and RNA levels (β-actin and GAPDH) was observed in two groups (4 °C and 15 °C), whereas the S type curve relationship between the expression levels of the two markers (β-actin and GAPDH) and PMI was observed in the 35 °C group. The partial linear relationship between 18S rRNA and PMI was observed in the groups (15 °C and 35 °C). CONCLUSION Skin could be a suitable material for extracting RNA. The RNA expression levels of β-actin and GAPDH correlate well with PMI, and these RNA markers of skin tissue could be additional indice for the estimation of PMI.
Actins ; Animals ; Autopsy ; Glyceraldehyde-3-Phosphate Dehydrogenases/genetics* ; Postmortem Changes ; RNA ; RNA Stability ; RNA, Ribosomal, 18S ; Rats ; Real-Time Polymerase Chain Reaction ; Regression Analysis ; Skin ; Temperature

OBJECTIVE: To investigate the process of apoptosis in lungs and liver induced by crushing hindlimbs of rat, and study the mechanism of crush injury. METHODS: The rat experimental model of hindlimbs crush injury was established. The cell apoptosis in lungs and liver was detected by TUNEL assay, and the expression of Bax, Bcl-2 and caspase-3 apoptin was examined by immunohistochemistry. RESULTS: Compared with the control group, the partial muscle injury of rat's hindlimbs was more serious with more apoptosis observed in lungs and liver (P < 0.05). The expression of Bax was up-regulated and Bcl-2 was down-regulated, whereas caspase-3 expression was activated (P < 0.05). CONCLUSION The cell apoptosis has increased significantly in lungs and liver after crush injury of hindlimbs in rat. The correlation factor released during tissue injury may mediate apoptosis process.
Animals ; Apoptosis/physiology* ; Caspase 3/metabolism* ; Genes, bcl-2 ; Hindlimb/injuries* ; Immunohistochemistry ; Liver/physiopathology* ; Lung/physiopathology* ; Rats ; Up-Regulation ; bcl-2-Associated X Protein