The cDNA encoding the precursor of cobrotoxin was cloned from the venom gland of the Chinese continental cobra ( Naja naja atra ) by RT-PCR. Its deduced amino acid sequence analysis showed that the mature protein was identical to that identified from the Taiwan cobra ( Naja naja atra ) by protein sequencing technique. The cDNA encoding the mature protein was then subcloned into the expression vector pMAL-P2. The gene of CMl 1, which was formed by ligation of the fragments of the synthetic oligonucleotides, was also cloned into the expression vector pMAL-P2. After induction of IPTG, both of the neurotoxins were overexpressed as soluble fusion proteins which were confirmed by SDS-PAGE and western blotting. The expressed fusion proteins was purified by sepharose 6B-amylose affinity chromatography and DEAE-sepharose FF chromatography. Both of the recombinant proteins achieved after digestion by factor Xa showed the in vivo toxicity.

Aim To compare high-content cell imaging system and other methods in detecting cell proliferation, including the traditional thymidine ( 3H-TdR) incorporation method, methyl thiazolyl tetrazolium ( MTT ) method and cell counting kit-8 (CCK-8) method. Methods The fibroblast-like synovial cells (FLS) were used as the study object to observe the sensitivity and stability of FLS proliferation in different methods by using the usual proliferative stimulant tumor necrosis factor-α ( TNF-α) and the known proliferation inhibitor methotrexate at different concentrations. Results The 3H-TdR method and high-content cell imaging could detect a significant inhibitory effect of 1 nmol ·L-1 MTX on FLS cell proliferation, MTT assay and CCK-8 method could detect the significant inhibitory effect of 10 nmol· L-1 MTX on FLS cell proliferation. 3H-TdR method was found to have a large degree of discretization in the data set, with a standard deviation of 32.61% ~61.36% , and the MTT method was 11.9% ~ 17.8% , the CCK-8 method was 17.15% ~32.88% , and the high-content cell imaging system method was 12.66% ~26.54%. Conclusion The method of high-content cell imaging system is more accurate and stable for detecting cell proliferation.

OBJECTIVETo explore the effect of acupuncture-drug compound anesthesia on immune function in patients with extracorporeal circulation undergoing cardiac surgery.

METHODSThirty cases undergoing cardiac surgery which included atrial septal defect neoplasty, ventricular septal defect neoplasty, mitral valve replacement and pulmonary valve coarctotomy were randomly divided into group A and group B, 15 cases in each group. Group A was given general anesthesia plus acupuncture at Neiguan (PC 6), Lieque (LU 7) and Yunmen (LU 2), and group B was given simple general anesthesia. Tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2) and interleukin-10 (IL-10) levels before and after surgery were compared.

RESULTSThe level of TNF-alpha was increased and the levels of IL-2 and IL-10 in the serum were decreased in both groups after extracorporeal circulation for 2 h and 24 h, and the ranges of all changes were more less in group A (all P < 0.05).

CONCLUSIONCompared with simple general anesthesia, acupuncture-drug compound anesthesia can improve immune suppression partially in the perioperative periods under the same conditions of controlling anesthesia degree.

Acupuncture Analgesia ; Adult ; Anesthesia, General ; Cardiac Surgical Procedures ; Female ; Heart Diseases ; blood ; immunology ; surgery ; Humans ; Inflammation Mediators ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Male ; Middle Aged ; Perioperative Care ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

Objective To detect Human Cytomegalovirus (HCMV) DNA in urine samples from the following groups:pregnant women,sick newborns,hospitalized nephropathy patients,renal transplant recipients and normal population.Preliminarily study the relationship of HCMVinfection and renal disease.Methods To detect HCMV DNA in morning urine samples by Real-time fluorescence quantitative PCR (FQ-PCR).Results The positive rates of HCMV DNA in the urine of pregnant women,sick newborns,hospitalized nephropathy patients,renal transplant recipients and normal population are 8.1 8%,3.45%,18.54%,25.42%,0.56%.Conclusion The infection rates of HCMV in the urine of pregnant women and sick newborns are very high in Guangxi,it should take serious measures to prevent and control the situation.HCMV probably participate in the iniury of kidney,and worsen the disease.It should be one of the causes of renal disease.

Objective To identify and verify the distribution of Telocytes derived from heterogeneous interstitial cells in the vital organs of ApoE mice.Methods Heart,kidney,and liver tissues were harvested from ApoE adult mice. Immunohistochemical assays were performed by using different immunobiological markers.Results Telocytes were found in these vital organs. The expressions of immunobiological markers differed among different organs. CD34,CD117,and CD28 were positively expressed in Telocytes in cardiac tissue;CD117 and plateled-derived growth factor-Α were negatively expressed in Telocytes in renal tissue;and CD117 and plateled-derived growth factor receptor-Α had negative expression in Telocytes in hepatic tissue. Furthermore,the distribution of Telocytes also differed in the same organ.Conclusions Telocytes exist in the vital organs of ApoE mice,as demonstrated by immunohistochemisty assay. The expressions of immunobiological markers differ among Telocytes in different organs.
Animals ; Antigens, CD34 ; metabolism ; CD28 Antigens ; metabolism ; Kidney ; cytology ; Liver ; cytology ; Mice ; Mice, Knockout, ApoE ; Myocardium ; cytology ; Proto-Oncogene Proteins c-kit ; metabolism ; Telocytes ; cytology

OBJECTIVETo evaluate the long-term effect of sodium glycididazole (CMNa) as a hypoxic radiosensitizer on the radiotherapy for nasopharyngeal carcinoma.

METHODSBetween May 1999 and May 2002, 211 patients with pathologically confirmed nasopharyngeal carcinoma were randomized into group-A treated by radiotherapy plus CMNa or group-B by radiotherapy alone. The staging was determined according to 92' Fuzhou staging systerm. The type, procession and dosage of radiotherapy were identical in both groups. The early adverse effect grade was assessed based on the CTC2.0 criteria and the late adverse effects were evaluated according to the RTOG/EORTC criteria. The median follow-up time was 52 months. All the data was analyzed by the SPSS 13.0 software. Characteristics and adverse events of these patients were compared between the two groups using t-test and the Wilcoxin rank sum test. Time-to-event curves were estimated using the Kaplan-Meier method. The prognostic parameters were analyzed using univariate analysis and the Cox multivariate regression analysis.

RESULTSThe clinical data of the two groups were comparable. The 3-year survival was 88.4% in group-A, while 75.2% in group-B, with a statistically significant difference between two groups (P = 0.010). Univariate analysis showed that the 3-year survival was statistically correlated with N-staging ((N0-1, 86.9%, N2-3 73.8%, P < 0.001), T-staging (T1-2 85.6%, T3-4 79.3%, P = 0.014), TNM staging (P = 0.039), and whether using CMNa or not during rediotherapy (Group-A 88.4%, Group-B 75.2%, P = 0.010). The 5-year recurrence-free survival, 5-year metastasis-free survival and 5-year overall survival were 75.8%, 74.9% and 77.7% in Group-A, while 63.0%, 63.0% and 62.4% in Group-B with a statistically significant difference between two groups (0.013, 0.022 and 0.010, respectively). If stratified in the subgroups, the overall survival of stage III - IV patients was statistically different between group A and B (P = 0.009), however, not of stage I - II patients (P = 0.502). Cox multivariate regression analysis showed that the independent prognostic parameters for survival were N-stage (RR = 3.288) , T-stage (RR = 2.147) and use of CMNa during rediotherapy (RR = 0.407). However, there was no statistically significant difference between two groups in acute or late adverse effects on nervous system or heart, which suggested that use of CMNa during radiotherapy would not aggravate the toxicity caused by radiotherapy.

CONCLUSIONSodium glycididazole is well tolerable effective as a hypoxic radiosensitizer, which can improve the efficacy of radiotherapy and the long-term result of nasopharyngeal carcinom a patients, especially for the stage III - IV patients.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Male ; Metronidazole ; adverse effects ; analogs & derivatives ; therapeutic use ; Middle Aged ; Multivariate Analysis ; Nasopharyngeal Neoplasms ; pathology ; radiotherapy ; Neoplasm Staging ; statistics & numerical data ; Prognosis ; Proportional Hazards Models ; Radiation-Sensitizing Agents ; adverse effects ; therapeutic use ; Time Factors ; Treatment Outcome ; Vomiting ; chemically induced

BACKGROUND: Albuvirtide is a once-weekly injectable human immunodeficiency virus (HIV)-1 fusion inhibitor. We present interim data for a phase 3 trial assessing the safety and efficacy of albuvirtide plus lopinavir-ritonavir in HIV-1-infected adults already treated with antiretroviral drugs. METHODS: We carried out a 48-week, randomized, controlled, open-label non-inferiority trial at 12 sites in China. Adults on the World Health Organization (WHO)-recommended first-line treatment for >6 months with a plasma viral load >1000 copies/mL were enrolled and randomly assigned (1:1) to receive albuvirtide (once weekly) plus ritonavir-boosted lopinavir (ABT group) or the WHO-recommended second-line treatment (NRTI group). The primary endpoint was the proportion of patients with a plasma viral load below 50 copies/mL at 48 weeks. Non-inferiority was prespecified with a margin of 12%. RESULTS: At the time of analysis, week 24 data were available for 83 and 92 patients, and week 48 data were available for 46 and 50 patients in the albuvirtide and NRTI groups, respectively. At 48 weeks, 80.4% of patients in the ABT group and 66.0% of those in the NRTI group had HIV-1 RNA levels below 50 copies/mL, meeting the criteria for non-inferiority. For the per-protocol population, the superiority of albuvirtide over NRTI was demonstrated. The frequency of grade 3 to 4 adverse events was similar in the two groups; the most common adverse events were diarrhea, upper respiratory tract infections, and grade 3 to 4 increases in triglyceride concentration. Renal function was significantly more impaired at 12 weeks in the patients of the NRTI group who received tenofovir disoproxil fumarate than in those of the ABT group. CONCLUSIONS: The TALENT study is the first phase 3 trial of an injectable long-acting HIV drug. This interim analysis indicates that once-weekly albuvirtide in combination with ritonavir-boosted lopinavir is well tolerated and non-inferior to the WHO-recommended second-line regimen in patients with first-line treatment failure. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT02369965; https://www.clinicaltrials.gov.Chinese Clinical Trial Registry No. ChiCTR-TRC-14004276; http://www.chictr.org.cn/enindex.aspx.
Adult ; Anti-HIV Agents/adverse effects* ; Antiretroviral Therapy, Highly Active ; China ; Drug Therapy, Combination ; HIV Infections/drug therapy* ; HIV-1 ; Humans ; Maleimides ; Peptides ; Ritonavir/therapeutic use* ; Treatment Outcome ; Viral Load