Adult ; Cholesterol ; Dilatation ; Ear Canal ; Ear Neoplasms ; surgery ; Ear, Middle ; pathology ; Eustachian Tube ; Female ; Granuloma ; surgery ; Humans ; Male ; Mastoidectomy ; Middle Aged ; Treatment Outcome

Chinese cobra (Naja naja atra) cardiotoxins are three-fingered family with 60～62 amino acids bind by four disulfide bonds. CardiotoxinⅢ (CTXⅢ) is one of the major toxic component which can cause hemolysis and cytotoxicity. However, there is no report on the fusion expression of CTXⅢ in soluble form so far. The cloning, expression and purification of recombinant CTX Ⅲ (rCTXⅢ) from Naja naja atra in E. coli and in yeast Pichia pastoris were reported here. CTXⅢ gene, fused with enterokinase in E.coli His-patch Thioredoxin expression system, were expressed in soluble form and released by osmotic-shock treatment. CTX Ⅲ gene was also cloned and expressed in the methylotropic yeast Pichia pastoris pPIC9K expression vector in the first time. The yield of the secretion level was 9.5 mg/L. Using straightforward one-step chromatography procedure, the rCTXⅢ, with three additional amino acids (GYT) at the N-terminal site, was purified to a purity of more than 90% and recovery yield of 65%. The purified rCTX Ⅲ was further characterized by cytotoxic assay with IC50 4.66μg/ml. An effective expression and purification system for recombinant CTXs in P. pastoris was developed, this system will permit us the ready isolation of active cardiotoxins. This protocol can also be easily used for the production of the toxin in a larger scale with low cost.

Objective:To investigate the status of the vestibular function of the patients with chronic positional symptoms after peripheral acute vestibular syndrome (AVS) and the curative effect of the vestibular rehabilitation therapy (VRT). Method:Using caloric test (CT), head shaking nystagmus test (HST), cervical vestibular evoked myogenic potentials as well as ocular vestibular evoked myogenic potentials to estimate the function of semicircular canal and otolith organs. The patients with normal VEMPs are divided as Group A. Otherwise are as Group B. Both groups are treated with VRT. The curative effect is estimated by vestibular symptom index (VSI) and Berg balance scale (BBS). Result:Thirty-three of 37 patients (86.5%) had an abnormal result of CT and HST, with 23 of these patients (65.7%) had an abnormal of both test. Twenty-two patients (59.5%) were in Group A and 15 (40.5%) in Group B. Before the therapy, Group B had a higher score of the balance and dizziness symptoms of VSI (P<0.05), and Group A had a higher score of the BBS (P<0.05). After the therapy, the VSI scores of both groups dropped and scores of the BBS raised. Conclusion:Patients with chronic positional symptoms after peripheral AVS have dynamic vestibular lesions to different extents. Those with otolith organs lesions tend to have a worse function of balance. Nevertheless, patients have a better off after VRT.

OBJECTIVE: To investigate the efficacy of bone marrow mesenchymal stem cells (BMMSC) on children with refractory graft-versus-host disease (GVHD) and to judge the efficacy of BMMSC by dynamically monitoring the changes of cytokines in children with GVHD before and after infusion of BMMSC, so as to provide a theoretical basis for clarifying the mechanism of BMMSC. METHODS: 17 children with refractory aGVHD including 7 of grade II, 6 cases of grade III and 4 cases of grade IV after allo-HSCT were enrolled. All the children with aGVHD, who received routine immunosuppressive therapy, but the state of disease not improved, were treated with immunosuppressive drugs combined with BMMSC infusion. Study endpoints included safety of BMMSC infusion, response to BMMSC, and overall response of aGVHD. The serum levels of IL-2α, IL-6, IL-10, IL-8 and TNF-α in aGVHD patients were measured by chemiluminescence before infusion of BMMSCs and Day 7, Day 14 after infusion of BMMSCs. RESULTS: The cumulative median dose of BMMSCs was 5.5 (3.4-11.1) × 10/kg for average of 3.7 times, and the median time of 16.5 (4-95) days for the first infusion of MSCs. In 17 cases of refractory GVHD, 14 responded to treatment, whereas 3 patients failed. The total effective rate was 82.4% and no adverse reactions occurred. Of the 14 survived cases (82.4%), the median follow-up time was 944 (559-1245) days from the first infusion of MSCs. The levels of TNF-α in children with grade II, III and IV GVHD before treatment were 9.5±4.3 pg/ml, 16.3±10.9 pg/ml and 35.8±21.2 pg/ml respectively. The difference between grade II and IV, III and IV was statistically significant (P<0.05). Compared with the ineffective group of BMMSC infusion, the serum TNF-αlevel in the BMMSCs treatment effective group was 10.8±5.6 pg/ml vs 40.6±14.8 pg/ml (t=-3.901, P<0.05) before treatment. In the effective group of BMMSCs infusion, IL-10 20±17.4 pg/ml of day 14 was significantly higher than that 7.3±3.1 pg/ml before the treatment (t=-2.850, P<0.05), while , the serum levels of IL-2α, IL-6, IL-8, TNF-α were not statistically significantly different (P＞0.05). CONCLUSION The infusion of BMMSC is safe and effective in the treatment of refractory GVHD in children. TNF-αlevel relates with the severity of GVHD. BMMSC may play an anti-GVHD role by up regulating the level of cytokine IL-10 in vivo.

【Objective】 To evaluate the analgesic efficacy of erector spinae plane block(ESPB) in patients undergoing robot-assisted partial nephrectomy(RAPN) . 【Methods】 One hundred patients scheduled for RAPN were randomized into ESPB group(Group E) and thoracic paravertebral block(TPVB) group(Group T), with 50 cases in each group. Ultrasound-guided nerve block was performed by senior experienced anesthesiologists in both groups. All patients received 0.5% ropivacaine hydrochloride and postoperative patient controlled intravenous analgesia(PCIA). The imaging time, needling time, number of needle passes, difficulty scale, consumption of remifentanil and propofol, pain visual analog scale(VAS) scores at rest and movement within 24 hours after surgery, pressing frequency of PCIA pump, postoperative complications and patients' satisfaction with analgesic efficacy were assessed and recorded. 【Results】 Compared with TPVB, ESPB required significantly shorter imaging time and needling time, significantly fewer number of needle passes and lower difficulty scale(P < 0.001). No significant difference was observed in term of the rest parameters between two groups(P > 0.05) . 【Conclusion】 Pre-emptive ultrasound-guided ESPB results in similar analgesic efficacy with TPVB in patients undergoing RAPN. ESPB may be easier for the junior doctors to perform due to its apparent superiority.

OBJECTIVETo investigate the serum levels of cholesterol, triglyceride and high density lipoprotein cholesterol in children aged 3-14 years and its influencing factors.

METHODSThe cross-sectional survey study population was a representative sample from Guangdong province obtained by multi-stage randomized cluster sampling. Serum lipids in 6188 children aged > or = 3 years were assayed using automatic biochemical instrument. The data of social and demographic status were collected by face-to-face interview, and height and weight were obtained by physical examination.

RESULTSThe age-standardized and region-weighted means of serum total cholesterol (TC), triglyceride (TG), and high density lipoprotein cholesterol (HDL-C) were 0.80 mmol/L, 3.50 mmol/L and 1.28 mmol/L, respectively. For the mean of TG, there was no difference between metropolitan and middle city, nor between rich county and poor county. For TC, it was the highest in metropolitan, and there was no difference between rich and poor county. For HDL-C, the difference existed between every two regions. The age-standardized and region-weighted prevalence of high TG, high TC and low HDL-C were 2.2%, 2.1% and 8.0%, respectively. Metropolitan, rich county, low weight and age between 7.0-9.9 years are protecting factors for high TG, and the number of family between 3-4, age between 7.0-9.9, metropolitan, middle city and poor county are risk factors for high TC. Male, family income per year between 800-9999 RMB, middle city, rich county are protecting factors for low HDL-C.

CONCLUSIONThe prevalence of abnormal serum lipid was still low compared with other regions in China. The region, number of family member, age and sex may be the important factors influencing on serum lipid levels.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Cross-Sectional Studies ; Dyslipidemias ; epidemiology ; Female ; Humans ; Male ; Risk Factors ; Triglycerides ; blood

OBJECTIVETo describe the epidemiological characteristics of overweight and obesity among people aged 18 and over in Guangdong province in 2002, and to identify the populations and regions under high risk.

METHODSCross-sectional survey was used through sampling on multi-stage randomized clusters. Data of socialeconomic status were collected by face-to-face interview. Data on weight and height was obtained through physical check-ups.

RESULTSA sample size of 15 130 people and the mean body mass index (BMI) was 22.03 +/- 3.38 with no significant difference between males and females. However the significant difference was seen between cities and rural areas. The crude prevalence rate of overweight and obesity were 16.8% and 1.8%, and the age-adjusted rate were 15.0% and 1.7%, respectively. The crude rate of overweight in cities (24.8%) and males (17.5%) were higher than that in rural areas (9.4%) and females (16.2%). The crude rate of obesity in cities was seen higher than that in the rural areas, but not significantly different between females and males. The results of logistic regression analysis showed that the major risk factors influencing overweight would include household income, age, gender, smoking habits, physical exercises and location of residence.

CONCLUSIONAbout one sixth of the citizens in Guangdong province were considered to be overweighted and obesive had become an important public health problem. Integrated measures must be taken for prevention and control.

Adolescent ; Adult ; Body Mass Index ; China ; epidemiology ; Cross-Sectional Studies ; Diet ; Exercise ; Female ; Humans ; Male ; Obesity ; epidemiology ; Overweight ; Prevalence ; Risk Factors ; Sampling Studies ; Socioeconomic Factors ; Surveys and Questionnaires

OBJECTIVETo explore multi-drug resistance (MDR) of bladder cancer for the intravesical instillation.

METHODSUsing immunohistochemical staining, in 44-case human bladder cancer cells, the expressions of P-glycoprotein (P-gp), glutathione S-transferase (GST-pi) and topoisomerase (TOPO-II), were detected to find out the resistance to drugs.

RESULTSP-gp had a higher expression in 54.5% cases. GST-pi had no or a lower expression in 65.9% cases. TOPO-II had a higher expression in 29.5% but a lower expression in 65.9% cases.

CONCLUSIONDetecting the factors of MDR in bladder cancer cells could help to choose drugs for intravesical chemotherapy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Administration, Intravesical ; Adult ; Aged ; DNA Topoisomerases, Type II ; analysis ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Glutathione Transferase ; analysis ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Urinary Bladder Neoplasms ; drug therapy ; metabolism

OBJECTIVETo observe the effects of pravastatin on platelet-derived nitric oxide system in hypercholesterolemia (HC) and atherosclerosis (AS) in rabbits, and the relationship between these changes and atherosclerosis courses.

METHODSThirty male New Zealand white rabbits were randomly divided into three groups, 12 in group A, 12 in group B, and 6 in group C. All of them were fed daily with cholesterol-rich food during the first 12 weeks. In addition, in group A, pravastatin (10 mg) was orally administered daily. At the end of the 12th week, 6 in group A and B were killed randomly and their aortas were removed and the pathologic changes were observed. In the following 12 weeks, food enriched with cholesterol was substituted with normal food in all three groups. Pravastatin treatment was continued or started in the remaining members of group A and group B, but not in group C. At the end 24th week, all rabbits were killed and their aortas were examined for the fatty-streaks or atherosclerotic plaques. The expressions of endothelial NOS (eNOS) mRNA and inducible NOS (iNOS ) mRNA, NOS activity, NO production and the level of the serum lipids were measured at 0, 6th, 12th, 18th and 24th week.

RESULTSThe expression levels of platelet-derived NOS mRNA, eNOS mRNA ratio in group A had no difference at above time points, while in group B were reduced significantly at 6th week and 12th week compared with at 0 week (P <0.01), and increased at 18th week and 24th week compared with 12th week (P <0.05). The expression levels of eNOS mRNA in group C were reduced at 6th, 12th and 18th, 24th week compared with 0 week (P <0.05 and P <0.01, respectively), and were reduced in groups B and C compared with group A at 6th ,12th week (P < 0.05) and increased in group A and B compared with group C at 18th, 24th week (P <0.01). The expression levels of iNOS/mRNA among the three groups had no difference. Pathologic finding of the arteries: AS was not found in group A from the 12th to 24th week. While in group B, there were a lot of fatty-streaks on the entire intima of all large arteries at the 12th week. There were also fatty-streaks in the ascending aorta, but were improved at the 24th week. In group C, there were marked plaques in the entire aorta at the 24th week.

CONCLUSIONSThe expressions of platelet-derived eNOS mRNA, NOS activity, NO production are decreased in HC or AS rabbits. Pravastatin can up-regulate expressions of platelet-derived eNOS mRNA, NOS activity, leading to preventing or improving the pathological courses of AS.

Animals ; Atherosclerosis ; blood ; pathology ; Blood Platelets ; metabolism ; Disease Models, Animal ; Male ; Nitric Oxide ; blood ; genetics ; Nitric Oxide Synthase ; blood ; genetics ; Pravastatin ; pharmacology ; RNA, Messenger ; genetics ; Rabbits

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α+dendritic cells (DCs) in vitro was investigated in this study.Immature CD8α+ DCs were prepared from C57BL/6 (H-2b) bone marrow cells by using a cytokine cocktail.On the 3rd day of culture,CD8α- DCs were pulsed by allogeneic (Balb/c,H-2d) EL9611 leukemia antigen,or RM-1 syngeneic prostate cancer antigen,with the concentration series of 0,2.5,5.0,10.0,20.0 μg/mL,respectively,then antigen-loaded immature CD8α+ DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1∶1,2∶1and 4∶1.T cell proliferation was measured by MTT assay.Cytokines including interferon gamma (IFN-γ)and interleukin-10 (IL-10) in CD8α+ DCs and T co-culture supernatant were detected by using ELISA.Cytotoxic effect of antigen-specific T cells was tested by LDH release assay.Conventional mature DCs　(mDCs) induced from C57BL/6 (H-2b) bone marrow cells by using granulocyte-macrophage colony　stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control.The results showed that the　proliferative activity of T cells stimulated by CD8α+ DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α+ DC/T ratio increased (P＜0.05).When antigen concentration ≤ 5μg/mL and CD8α+ DC/T ratio ≤ 2∶1,the ability of CD8α+ DCs to stimulate T cell proliferation was　higher than mDC control in allogeneic tumor antigen-pulsed groups (P＜0.05),but not in syngeneic tumor antigen-pulsed groups (P＞0.05).The level of IFN-γ and IL-10 in CD8α+DCs and T cell co-culture　supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P＜0.05),and the　cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when　the CD8α+DC/T was 1∶1 or 2∶1 (P＜0.05).There existed a negative correlation between the level of　IL-10 and T cell proliferation.T cell cytotoxicity assay showed that when CD8α+ DCs were pulsed with　allogeneic tumor antigen,the maximal T cell killing efficiency could reach (100±7.7)％,whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)％.It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α+ DCs could stimulate T cells to exert the GVT effect in vitro,and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen.The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α+ DC/T　ratio (1∶1 and 2∶1).