Objective To express and purify recombinant and biologically active Clostridium difficile toxin B (rTcdB). Methods The genes of TcdB were amplified by polymerase chain reaction (PCR) using chromosomal DNA from a toxigenic strain, and cloned into a shuttle vector pHis1522.The sequences of TcdB genes in the vector were verified by DNA sequencing. The construction was transformed into Bacillus megaterium protoplasts and the protein expression was driven by a xylose promoter. The purified protein was tested for biological activity. Results rTcdB was successfully purified from bacterial crude extracts. Approximately 5-10 mg of highly purified recombinant toxin was obtained from one liter of bacterial culture. The expressed rTcdB had molecular mass similar to the native toxin, and its biological activity was proved to be similar to its native counterpart after an extensive examination. Conclusion rTcdB with biological activities is successfully expressed in Bacillus megaterium.

AIM: The auxology-related indexes are important for tissue construction. In this article, we investigated the incidence of overweight and obesity and their effects on physical functions and qualities of Guangzhou adults in 2005. METHODS: 2 056 adults including 1 027 males and 1 029 females were randomly selected from Guangzhou in 2005 for physical examination. The anthropometric data such as body mass and height were collected to calculate body mass index (BMI). The subjects were divided according to the BMI criteria recommend by Group of China Obesity Task Force, and their physical functions and qualities were analyzed. RESULTS: In 2005, the detection rate of overweight and obesity in adult males were 28.7% and 5.2%, and in females were 18.6% and 5.3%. The values of diastole pressure and systolic pressure were increased with BMI, but the change tendency of step index and vital capacity/body mass was decreased with BMI. The rate of hypertension was increased with BMI, and male subjects had higher rate of hypertension than that of female subjects in all groups. Obese adult had the poorest values in vertical jump and sit-and-reach test. Those people with normal BMI had better balance ability than that of overweight and obese people. CONCLUSION: The rates of overweight and obesity in Guangzhou adult show a high level. The physical functions and qualities may be improved when the body mass increases or decreases in a certain range, but obesity has impact to those values.

Background: Although the erector spinae plane block has been used in various truncal surgical procedures, its clinical benefits in patients undergoing spinal surgery remain controversial. The aim of this meta-analysis was to evaluate the clinical benefits of erector spinae plane block in patients undergoing spinal surgery. Methods: We searched the Cochrane Library, PubMed, EMBASE, and China National Knowledge Infrastructure for randomized controlled trials comparing the erector spinae plane block with a nonblocked control for spinal surgery. Results: Twelve studies encompassing 696 subjects were included in our systematic review and meta-analysis. We found that the erector spinae plane block decreased postoperative pain scores and opioid consumption in the postoperative and intraoperative periods. Moreover, it prolonged the time to the first rescue analgesic, reduced the number of patients who required rescue analgesia, and lowered the incidence of postoperative nausea and vomiting. However, it did not exhibit efficacy in decreasing the incidence of urinary retention and itching or shortening the length of hospital stays, or the time to first ambulation. Conclusions Erector spinae plane block improves analgesic efficacy among patients undergoing spinal surgery compared with nonblocked controls; however, there is insufficient evidence regarding the benefits of erector spinae plane block for rapid recovery.

Background: Although the erector spinae plane block has been used in various truncal surgical procedures, its clinical benefits in patients undergoing spinal surgery remain controversial. The aim of this meta-analysis was to evaluate the clinical benefits of erector spinae plane block in patients undergoing spinal surgery. Methods: We searched the Cochrane Library, PubMed, EMBASE, and China National Knowledge Infrastructure for randomized controlled trials comparing the erector spinae plane block with a nonblocked control for spinal surgery. Results: Twelve studies encompassing 696 subjects were included in our systematic review and meta-analysis. We found that the erector spinae plane block decreased postoperative pain scores and opioid consumption in the postoperative and intraoperative periods. Moreover, it prolonged the time to the first rescue analgesic, reduced the number of patients who required rescue analgesia, and lowered the incidence of postoperative nausea and vomiting. However, it did not exhibit efficacy in decreasing the incidence of urinary retention and itching or shortening the length of hospital stays, or the time to first ambulation. Conclusions Erector spinae plane block improves analgesic efficacy among patients undergoing spinal surgery compared with nonblocked controls; however, there is insufficient evidence regarding the benefits of erector spinae plane block for rapid recovery.

Objective To develop an ELISA(Enzyme-Linked Immunosorbent Assay)diagnostic kit for early rapid detection of sarum anti-EV71 antibody and evaluate its clinical application value.Methods Recombinant protein VP1 of EV71 were prepared and purified as an immobilized antigen for establishment of an indirect ELISA for detection of serum anti-EV71 IgM and anti-EV71 IgG.Compared with RT-PCR.isolation of EV71 and micro-neutralizing assay.the clinical application value of anti-EV71 IgM and anti-EV71 ISG in the diagnosis of EV71 disease was evaluated.Results In comparison with RT-PCR.the sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgM antibody were 83%,85%,81%and 87%,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgG antibody were 72%,74%,68%and 77%.respectively.Compared with viral isolation assay.the sensitivity and specificity of anti-EV71 IgM antibody were 85%and 97%,respectively.The sensitivity and specificity of anti-EV71 IsG antibody were 75%and 77%,respectively.In addition.the titers of anti-EV71 IgG antibody were significantly correlated with the titers of neutralizing antibody to EV71 by linear regression analysis(r=0.72,P＜0.05).Finally,the serum titers of anti-IgG from patients with EV71 associated hand food and mouth disease at convalescent stage exhibited significantly higher than that of the same patients at acute stage(P＜0.01),but the titers of anti-IgM had no significant difference(P＞0.05).Conclusions With VP1 recombinant protein used as an immobilized antigen,an indirect ELISA diagnostic kit was successfully develooed for detection of serum anti-human EV71 IgM and anti-human EV71 IgG antibodies.

Objective To investigate the phenotype, frequency of Th17 cells and the association between Th17 cells and viral clearance in patients with H1N1 influenza A. Methods Three groups including 70 confirmed patients with H1N1 influenza A, 30 patients with seasonal influenza as well as 68 healthy subjects as controls were enrolled in this study. The percentages of Th1, Th2, Treg and Th17 lymphocytes in the peripheral blood were determined by intracellular staining and flow cytometry. The levels of interferon-γ (IFN-γ), transforming growth factor-beta (TGF-β),interleukin-6 (IL-6) in plasma and supernatant of the peripheral blood mononuclear cell (PBMC)culture were quantified by enzyme-linked immunosorbent assay (ELISA). Viral load in nasopharyngeal swabs was detected by real time quantitative reverse transcription-polymerase chain reaction (RTPCR). Data were analyzed by one way ANOVA and liner correlation analysis. Results The percentage of Th17 cells in H1N1 influenza A patients was (2. 740±0. 210)%, which the percentage of was significantly decreased compared to healthy subjects (3. 443 ±0. 154)% and seasonal influenza patients (3. 443±0. 277) % (F=4. 242, P＜0. 05); while the percentage of Thl, Th2 and Treg cells were not significantly different among these groups. Moreover, the TGF-β level in plasma of H1N1 influenza A patients was (10±8) ng/mL, which was significantly lower than healthy subjects (43 ±32 ) ng/mL and seasonal influenza patient ( 18 ± 10) ng/mL ( F= 17.72, P＜0.01 ). The TGF-β level in the supernatant of PBMC culture of H1N1 influenza A patients was (782 ± 736) pg/mL, which was significantly lower than healthy subjects (1462±315) pg/mL and seasonal influenza patients (1481 ±348) pg/mL (F=5. 730, P＜0.01). Additionally, the viral clearance period was inversely correlated with the percentage of Th17 cells (r=-0.38, P=0.02). Conclusions The proportion of Th17 cells in patients with H1N1 influenza A is significantly decreased, which is closely correlated with the level of TGF-β. This decrease may results in the delayed viral clearance.

Objective To Screen for safe and effective vaccine candidates for EV71,provide a theoretical basis for development of EV71 vaccines in the future.Methods VP1 gene of enterovirus was used to design a target for development of EV71 vaccines.Different vaccine candidates,including inactivated EV71 vaccines,VP1 protein vaccine,DNA vaccines of different doses,were used to challenge female BALB/c mice by intramuscular injection at baseline (0),2 weeks,4 weeks,and caudal vein blood was collected at 0,2,4,6,8,10,and 16 weeks,and BALB/c mice were sacrificed and the spleen cells were collected for detection of both humoral immunity and cellular immunity to evaluate the efficacy and safety of the vaccine candidates.Results IgG antibody titers were increased at 2 weeks,remarkably increased at 4 weeks,reached a peak at 8 weeks,at least sustained for 16 weeks during the whole observation period,subtypes of IgG1 and IgG2a were the major component.The three vaccines could induce cellular immunity characterized by EV71 specific γ-IFN and IL-4 production.Our results indicated that inactivated EV71 vaccine was superior to the other vaccine candidates.Conclusions Inactivated EV71 vaccines,VP1 protein vaccine,DNA vaccines can induce both strong and sustainable humoral and cellular immunities in challenged mice,and the inactivated EV71 vaccine is superior to the other vaccine candidates,which needs to be proved their immunity by challenge assay in the future.

Purpose:To evaluate the surgical method of pheochromocytoma.Methods:Data of 103 cases of patients with pheochromocytoma was reviewed retrospectively.Results:The operation of 49 patients with Pheochromocytoma were performed by the 11 th rib intercostal incision, 18 cases by the 10th rib intercostal incision, 15 cases by transrectus incision, 9 cases by abdominothoracic incision, 5 cases by abdominal subcostal oblique, 3 cases by inferior abdominal incision, 3 cases by posterolateral incision. 1 patient received the laparoscopic surgery.Conclusions:① Most surgical pathway of the pheochromocytoma in adrenal can be performed by the 11th rib intercostal incision of the 10th rib intercostal incision.②The suitable pathways for the giant pheochromocytoma adhering to big vessels are transrectus incision or abdominothoracic incision.③ For the mutiple masses of extra-adrenal pheochromocytoma, the middle abdominal incision may be the choice.