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.

OBJECTIVETo investigate the radiosensitizing effect and its mechanism of 3-MA in human hypopharynx cancer cells.

METHODS5 mmol/L of 3-MA combined with 2 Gy or 4 Gy of X-ray was utilized to deal with Fadu cells, and the cell livability (cloning efficiency) and DNA lesion severity (tail moment) of each groups was examined by clonogenic survival assay and comet assay, then differences were compared between groups by independent-sample T test. Fadu cells were then treated with different dose of 3-MA (1, 2, 5, 10 mmol/L), the alteration of cell cycle distribution was detected by flow cytometer, and differences among groups were analyzed through one-way analysis of variance. The expression of p62 and cyclinB1 in each group was examined by western blot.

RESULTSThe livability and DNA lesion severity of cells treated with 3-MA alone showed no notable variation. Compared with non-3-MA groups, the cloning efficiency of cells treated with 3-MA decreased much more after irradiated with 2 Gy or 4 Gy of X-ray (t = 13.41 or 13.98, P < 0.001), and the cells showed a more serious DNA lesion (t = 7.07 or 6.91, P < 0.001). The G2/M percentages of cells in the control group and groups treated with 1, 2, 5, 10 mmol/L of 3-MA were 17.10 ± 1.20, 23.30 ± 2.3, 39.90 ± 3.12, 58.47 ± 1.65, 76.13 ± 3.51 and differences among groups were statistically significant (F = 278.4, P < 0.05). The expression of p62 in cells treated with 3-MA showed a dose-dependent increase, while cyclinB1 showed a dose-dependent decrease.

CONCLUSIONSThe autophagy inhibitor 3-MA could enhance radiosensitivity of human hypopharynx cancer cells by inducing G2/M arrest and enhancing irradiation-induced DNA damage.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenine ; analogs & derivatives ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cyclin B1 ; metabolism ; DNA Damage ; drug effects ; Humans ; Radiation Tolerance ; drug effects ; Radiation-Sensitizing Agents ; pharmacology ; Sequestosome-1 Protein

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 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 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 investigate and compare the short-term outcome of patients with low-middle frequency sudden deafness treated with alone or combination treatment.

METHODSFrom August 2007 to October 2011, 205 patients with the diagnosis of low-middle frequency sudden deafness who were from 33 different clinical centers were recruited. All patients were followed up for four weeks from the initial examination. Patients were treated with steroid , Ginaton, batroxobin respectively, or Ginaton and steroid combination treatment.

RESULTSThe total effective rate was 90.73%. In Ginaton group, the total effective rate was 87.27%, 89.19% in steroid group, 87.80% in batroxobin group, and 95.83% in Ginaton and steroid group. Considering the total effective rate, there was no statistical difference between four groups (χ(2) = 7.98, P = 0.54). The clinical cure rate for steroid alone was 81.01%, Ginaton alone 76.36%, batroxobin alone 68.29%, and Ginaton and steroid combination treatment 80.56%. There were no clinically significant differences between the different treatments (P > 0.05).

CONCLUSIONSThe low-middle frequency sudden deafness tends to have a relatively favorable prognosis. The steroid played a good effect in the treatment. But different treatments either improving the microcirculation of inner ear or alleviating edema blood has undifferentiated results. Therefore the combination therapy may be more effective.

Batroxobin ; China ; epidemiology ; Combined Modality Therapy ; Drug Therapy, Combination ; Hearing Loss, Sensorineural ; Hearing Loss, Sudden ; classification ; diagnosis ; epidemiology ; therapy ; Humans

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