Objective To explore the characterization of mitochondrial DNA and cytochrome b(Cytb) gene from Martes zibellina L..Methods The mtDNA were extracted from fresh heart and liver of Martes zibellina L.by alkaline lysis method respectively,and Cyt b gene segment was amplified by PCR technique and sequenced.Results The complete 16800 bp mtDNA was separated from both samples,and the Cytb gene with 310 bp segments was amplified by PCR and the sequence indicated that the homologous similarity was 99% with sable Martes zibellina L.(GenBank:AB026109.1) and it was 45% with other animal similarity.Conclusion There is complete mtDNA in tissues of Martes zibellina L.and Cytb partial gene from martes zibellina L.may be used as the genetic marker for identification of different species.

Objective To study the relationship between lymph node micrometasis and clinicopathological parameters.Methods Immunohistochemical method was adopted to detect cytokeratin pan(CK)、Epithelial Membrane Antigen(EMA) and Carcinoembryonic Antigen(CEA) in 389 lymph nodes from 36 cases of breast cancer, in which conventional pathological diagnosis showed no lymph node metastasis.Results Micrometastasis was found in 38 lymph nodes (9.2) of 12 cases (33.3) .The incidence of micrometasis was correlated with tumor size. Pathological diagonosis and metastasis lymph nodes. Conclusion Micrometastasis detection in negative lymph nodes of breast cancer is recommended to precisely determine the tumor stage, in order to direct cancer therapy and predict prognosis.

OBJECTIVETo obtain recombinant baculovirus containing full-length structural gene of HEV and to express HEV structural protein.

METHODSFull-length structural gene of HEV (5147-7126 nt) was inserted into baculovirus expression vector pAcUW51. The recombinant plasmid and Baculo Gold DNA were co-transfected into insect cell line Sf9. Single plaque was picked and amplified and expression of HEV structural protein was tested by SDS-PAGE, Western blot and immunofluorescence methods.

RESULTSSDS-PAGE analysis showed HEV structural protein was highly expressed; Western blot and immunofluorescence assay showed that the expression product could specifically react with HEV positive sera, confirming the protein possessing HEV specific antigenicity.

CONCLUSIONSHEV structural protein was successfully expressed using baculovirus expression system.

Animals ; Baculoviridae ; genetics ; Cells, Cultured ; Gene Expression ; Genes ; genetics ; Genetic Vectors ; Hepatitis E virus ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Viral Structural Proteins ; biosynthesis ; genetics

Coronary heart disease is one of the leading causes of death worldwide. Advanced age is associated with increased prevalence, morbidity and mortality of coronary heart disease. Older adults are at higher risk of complications and accelerated physical deterioration following cardiovascular events than youn-ger patients. In recent years, researchers have proposed that exercise-based cardiac rehabilitation is an im-portant part of the treatment of patients with coronary heart disease, especially contributing to the improve-ment of the quality of life of elderly patients. This review summarizes the current status and influence of car-diac rehabilitation on elderly patients with coronary heart disease.

BACKGROUNDTo observe the protective effect of hepatitis E virus (HEV) ORF2 recombinant protein expressed in prokaryote cell cynomolgus macaques (cynos) against challenging with wild-type HEV.

METHODSCynos were immunized with HEV ORF2 recombinant protein and then challenged with wild-type HEV, the unimmunized cynos were used as control. Blood samples were collected and tested to see if there were dynamic changes of ALT and antibody to HEV before and after challenge with wild-type HEV.

RESULTSAll the five unimmunized cynos re-presented hepatitis 3 weeks after challenging with wild-type HEV. However, all the five immunized cynos showed no hepatitis and pathological changes.

CONCLUSIONSCynos can be efficiently protected by immunization with HEV ORF2 recombinant protein against wild-type HEV. This protein can be a promising candidate for HEV vaccine.

Animals ; Female ; Hepatitis Antibodies ; blood ; Hepatitis E ; immunology ; prevention & control ; Hepatitis E virus ; immunology ; Macaca mulatta ; Male ; Recombinant Proteins ; immunology ; Viral Proteins ; immunology

BACKGROUNDTo obtain thermal stable, soluble, biologically active hepatitis E virus recombinant antigen using thioredoxin fusion expression system.

METHODSHEV ORF2 gene fragment (6964-7126 nt) was inserted into thioredoxin fusion expression vector pThioHisA. The recombinant plasmid was transformed into E. coli BL21 strain. After induction with IPTG, cells were lysed and the supernatant was subjected to 80 degree treatment for 10 minutes. After centrifugation, the supernatant was tested by ELISA.

RESULTSSDS-PAGE analysis showed the thioredoxin. HEV fusion protein was highly expressed and was thermally stable, soluble. HEV specific ELISA confirmed this fusion protein possessing HEV specific antigenicity.

CONCLUSIONSUsing thioredoxin fusion expression system, a soluble, thermal stable, biologically active HEV recombinant antigen was successfully expressed.

Antigens, Viral ; biosynthesis ; genetics ; Gene Expression ; Genetic Vectors ; Hepatitis E virus ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Thioredoxins ; genetics ; Viral Proteins ; biosynthesis ; genetics

BACKGROUNDTo get early laboratory study of type specific antigenicity of herpes simplex virus II.

METHODSPCR and prokaryotic expression technique.

RESULTSHerpes simplex virus II type specific gene fragment was expressed in E.coli and the products can be used as specific antigen for the detection of anti\HSV in the recovery sera.

CONCLUSIONSCloning and express of HSVII type specific antigen found the basis for developing specific diagnosis methods and vaccine of HSV.

Antigens, Viral ; immunology ; Cloning, Molecular ; Gene Expression ; Herpesvirus 2, Human ; genetics ; Humans ; Immunoglobulin G ; blood ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology

Objective: To investigate the correlation between CXC chemokine receptor 6 (CXCR6) and prognosis of renal clear cell carcinoma. Methods: A total of 100 patients with renal clear cell carcinoma who underwent surgery in the First People's Hospital of Ziyang from January 2013 to October 2014 were selected. Immunohistochemistry was used to detect the expression of CXCR6 in 100 cases of renal clear cell carcinoma and adjacent normal tissues. The patients were followed up to observe the positive rate of CXCR6 expression and the factors affecting overall survival (OS) and progression-free survival (PFS) of renal clear cell carcinoma and adjacent normal tissues. Results: The positive rate of CXCR6 expression in renal clear cell carcinoma tissues was higher than that in adjacent tissues, and the difference was statistically significant [46.0% (46/100) vs. 20.0% (20/100), χ² = 15.287, P < 0.01]. The positive expression rate of CXCR6 in patients with clinical stage Ⅲ-Ⅳ, lymph node metastasis and pathological grade Ⅲ-Ⅳ was higher than that in patients with clinical stage Ⅰ- Ⅱ, pathological grade Ⅰ- Ⅱ and without lymph node metastasis, and the difference was statistically significant (all P < 0.05). A total of 92 patients were followed up and 40 died. The follow-up time reached to (35-60) months and the median follow-up time was 48.9 months. Kaplan-Meier and log-rank test showed that patients with positive CXCR6 expression had lower 3-year OS and PFS rate compared with patients with negative CXCR6 expression, and the difference was statistically significant (3-year OS rate: 52.1% vs. 78.6%, χ² = 10.027, P = 0.001; 3-year PFS rate: 48.3% vs. 67.8%, χ² = 4.344, P = 0.037). Cox regression multivariate analysis showed pathological grade (OS: OR = 2.154, 95% CI 1.547-9.517, P = 0.023; PFS: OR = 1.235, 95% CI 1.109-5.917, P = 0.042), lymph node metastasis (OS: OR = 1.412, 95% CI 1.109-5.917, P = 0.041; PFS: OR = 1.841, 95% CI 1.354-8.994, P = 0.010), CXCR6 expression (OS: OR = 1.864, 95% CI 1.358-6.813, P = 0.031; PFS: OR = 1.457, 95% CI 1.127-6.884,P = 0.025) were independent risk factors of OS and PFS for renal clear cell carcinoma patients treated by the surgery. Conclusions The positive expression of CXCR6 is an independent risk factor for OS and PFS of renal clear cell carcinoma. The prognosis of patients with positive CXCR6 expression is poor.

Objective To compare the difference in dose of general anesthesia between pastoral patients and urban pa-tients.Methods The patients who performed abdominal surgery under general anesthesia in Peoples hospital of Bortala Mongo-lian Autonomous Prefecture from January 1st to April 30th 2022 were included and divided into pastoral patient group and ur-ban patient group according to the residential place of patients.Anesthesia and surgery were carried out according to hospital regulations.The general characteristics of all patients were recorded and the dosages of general anesthesia drugs were compared between the two groups.Results There was no significant difference between the two groups in general and intraoperative con-ditions.Patients'performances were stable,and no severe adveres reaction was observed.The dosages of Propofol,Remifen and Rocuronium in pastoral patients were higher than that in urban patients(all P＜0.05).Conclusion The dosage of general anes-thesia for patients in pastoral areas is higher,which ensures the stability of anesthesia depth and vital signs of patients,creates good conditions for surgery and promotes rapid recovery of patients.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.