Objective To express human thyroid peroxidase (hTPO) epitopes gene and apply its products in clinical assay. Methods hTPO epitopes gene was cloned into expression vector pGEX 4T 3 then transformed into E. coli BL21. Expression of hTPO gene was induced by isopropyl ? D thiogalactoside, expressed product (GST hTPO) was purified by affinity chromatography and their immunoactivity was demonstrated. ELISA technique using GST hTPO as antigen was established for determining TPOAb. Serum TPOAb level was determined, and HLA DR antigen, dendritic cells and lymphocytes in the thyroid gland tissue were observed in these same AITD patients.Results GST hTPO acquired from procaryotic expression had high purity and good immunoactivity. The CVs of the ELISA technique established with GST hTPO were between 5.93%～7.59%. A significantly positive correlation was found between the TPOAb levels determined respectively by ELISA and RIA method. Serum TPOAb level and distribution of HLA DR antigen and dendritic cells showed the same ascending tendency following the aggravation of lymphocyte infiltration. Conclusion Product of genetic engineering, GSH hTPO, can be used to establish a clinical assay for TPOAb. The correlation between the level of serum TPOAb and the detriment of thyroid tissue is demonstrated.

Cardiomyopathies ; therapy ; Humans ; Medicine, Chinese Traditional

The authors adopted acupuncture plus auricular electro-acupuncture to treat 38 cases of dysmenorrhea belonging to qi stagnation and blood stasis pattern, the better therapeutic effect was achieved, with the effective rate of 97.4%.

12 m/s group,n=70;cfPWV≤12 m/s group,n=49). Carotid atheromatous plaque was assessed by Doppler echocardiograph;blood pressure and hemoglobin A1c were measured. Results After adjustment for body mass index (BMI),sex,stroke and fasting blood glucose,logistic multivariable analysis showed that cfPWV associated with age,systolic blood pressure(SBP),pulse pressure,coronary artery disease,diabetes,carotid atheromatous plaque and hemoglobin A1c. Conclusion Arterial stiffness in elderly patients with essential hypertension was strongly positively associated with age,SBP,pulse pressure,coronary artery disease,diabetes,carotid atheromatous plaque and hemoglobin A1c.

Objective To investigate the expression levels of serum visfatin ,leptin and other indexes and their correlation and clinical significance in the patients with type 2 diabetes mellitus(T2DM ) .Methods Eighty-two age-matched and gender-matched cases of T2DM (T2DM group)and 71 cases of healthy subjects(health control group)were tested serum visfatin ,leptin ,insulin ,lipid and glycemia levels .Results The serum visfatin ,leptin ,triglycerides(TG) ,total cholesterol(TC) ,high-density lipoprotein choles-terol(HDL-C) and low density lipoprotein cholesterol(LDC-C) ,fasting plasma glucose ,fasting insulin(FINS) ,homeostasis model assessment of insulin resistance(OMA-IR) ,2 h FPG ,2 h FINS had statistically significant differences between the T 2DM group and the health control group(P< 0 .05);the stepwise regression analysis showed that visfatin were positively correlated with BMI , FINS ,HOMA-IR ,TC ,TG ,LDL-C (P<0 .05)and negatively correlated with HDL-C(P<0 .05);leptin were positively correlated with BMI ,FINS ,HOMA-IR(P<0 .05) .Conclusion Serum visfatin and leptin in the T2DM patients are hither than those in the healthy subjects and closely related with blood lipid ,blood glucose and insulin ,which may become the new diagnostic indexes of T2DM .

Objective To explore the relationship between cognitive impairment and arterial stiffness in elderly patients. Methods A total of 142 elderly patients were enrolled. Cognitive function was assessed by mini-mental state examination (MMSE) and arterial stiffness was assessed by pulse wave velocity (PWV). A full score on the MMSE was 30, and cognitive impairment was defined as a score less than 24. All subjects underwent the measurement of PWV and MMSE . The subjects were divided into 2 groups: 93 were assigned to the normal cognitive function group (MMSE score 24), and the remainders (n=49) were assigned to the cognitive impairment group (MMSE score 24). Results The PWV was significantly increased in the cognitive impairment group than in the normal cognitive function group [(13.3±2.4) m/s vs. (11.8±2.2) m/s, t=3. 775, P=0. 000]. Logistic regression analysis showed that the PWV was also independently and significantly associated with the MMSE score. Conclusions The increase of arterial stiffness is an important risk factor for impaired cognitive function in elderly patients.

Response Surface Methodology was applied to optimize the culture components for lipopeptide production by Bacillus natto TK-1. In the first step, two level factorial design of Plackett-Burman was used to evaluate the influence of six related factors. It showed that three factors playing the important roles in the medium, including peptone, yeast extract powder and CaCl_2. The path of steepest ascent was used to approach the optimal region of the fermentation conditions subsequently. In the third step, the concentrations of those three main factors were further optimized by using Box-Behnken and Response Surface Analysis. By solving the quadratic regression model equation, the optimal concentrations of the variables were determined as: peptone 1.73%, yeast extract powder 0.063 %, CaCl_2 1.385?10-4mol/L. Under the optimal culture conditions, the diameter of haemolysis zone increased 29.3 % than before. HPLC analysis showed the precise production of lipopeptide was 30.2% higher than preliminary culture. Furthermore, at three batches cultivation, the experiment values under the optimal conditions agreed with the predictive values. It showed that Response Surface Methodology was proper and a good choice for optimization.

Fermentation for Eicosapentaenoic Acid(EPA) production by Nitzschia laevis at various temperature between 10℃ and 30℃ was investigated and the dynamics characteristics during fermentation process were also analyzed.Based on the results,a varying temperature nursing method of two stage control strategy is proposed:During the first stage,which comprises the delay phase and the initial index phase,the temperature is maintained at 25℃;then the temperature is shifted to 20℃ and kept up till the end of the fermentation process.By this method,a EPA content of 6.0% and a yield of 291.60 mg/L have been gained.These are 24.07% and 18.81% higher than that of fixed temperature(25℃) fermentation,respectively.

This thesis aimed at the Bacillus natto TK-1 screened out from Natto.The lipopeptide was purified using Thin-Layer Chromatography(TLC),and investigated its anti-tumor activity.After acid precipitation and methanol extraction,the lipopeptide was separated on TLC.Then the authors get the monomer surfactin which molecular weight is 1036Da through the High performance liquid chromatography(HPLC),electro spray ionization-mass spectrometry(ESI-MS) and infrared(IR).MTT method was implied to testify the anti-tumor activity of the purified sample from TLC.The results indicated a concentration and time-dependent relationships.After 48h,their IC50 were 40 mg/L.The detection with inverted microscope fluorescence microscope displays that the surfactin will cause a series of Morphological changes to the cells.In TUNEL experiment,the authors noticed that surfactin has the ability to induce apoptosis,besides this inhibition shows an obvious time-dependent relationship.

The study was to develop cis-dichlorodiammineplatinum(DDP)-loaded formulations using a novel type of self-assembled compound composed of block copolymers synthesized by poly(?-glutamic acid)(?-PGA).For the potential of targeting liver cancer cells,D-galactose was conjugated on the prepared ?-PGA.In vitro,DDP can be released from the resulting conjugate in PBS:there was a burst release during the first 8 h,then followed by sustained release.DDP could be easily incorporated into poly(?-glutamic acid)-D-galactose esterifiable derivative through a covalent bond.The yield of DDP incorporation into the esterifiable derivative was 9.4%～10.2%.In vitro experiments conclusively established that the poly(?-glutamic acid)-D-galactose esterifiable derivative-Cisplatin Complex Compound(?-D+-DDP)was much less toxic to normal cell lines than DDP only.The surviving rate of cells treated with ?-D+-DDP compound is higher than those treated with free DDP.Also it has obvious antitumor efficiency on human liver tumor BEL-7402 cells.HE staining indicated that the ?-D+-DDP compound make the BEL-7402 apoptosis.These results indicated that the conjugation of DDP to the esterifiable derivative reduced its cytotoxicity activity,but retains its antitumor activity in vitro.In conclusion,the ?-D+-DDP compound could be used as a potential clinic antitumor drug.The ?-PGA obtained by fermentation can be used as a valuable drug carrier system.