Objective To compare the transfection rate of AAV2-EGFP and AAV2/1-EGFP to human skin fibroblast(HFB).Methods Two recombinant adeno-associated viruses(AAV) encoding enhanced green fluorescent protein(EGFP) were constructed and transfected to HFB at multiple of infection MOI ranging from 104 to 106.Twenty-four hours after the infection,the expression rates of EGFP on cultured HFB were assessed by flow cytometry and the transfected cells were observed under fluorescence microscope.The killing effect of the virus on the infected cells was assayed by MTT.Results The transfection efficiency of AAV2-EGFP was increased as MOI increased.When MOI was 104,the transfection efficiency of AAV2-EGFP was 7.68%?1.18%;When MOI was 105,that was 52.12%?1.59%;When MOI was 106,that showed no significant increase.However,the transfection efficiency of AAV2/1-EGFP showed no obvious changes at any MOI.Conclusion AAV2 is more efficient than AAV2/1 in transfecting HFB,but neither AAV has a high transfection efficiency.

OBJECTIVETo observe the therapeutic effects of Berberine Capsule (BC) on patients with mild hyperlipidemia.

METHODSTotally 102 mild hyperlipemia patients were recruited. All patients were suggested to have proper diet and physical activity as basic therapy for 1 month of run-in period. Totally 97 patients completed it. Then they were randomly assigned to the berberine group (the treatment group, 49 cases) and the placebo group (the control group, 48 cases). Patients in the treatment group took BC 300 mg, while those in the control group took placebo 300 mg, thrice per day for 3 successive months. Then placebos and BC were interrupted for 2 months (as washout period). All subjects received only diet control and physical activity during washout period. After washout period, placebos and BC were re-administered to all patients in the same way for 3 months. Body mass index (BMI), fasting plasma glucose (FPG), TG, TC, LDL-C, and HDL-C were assessed after run-in period, washout period, at month 1, 2, 3 after the first therapy, at month 1, 2, 3 after second treatment, respectively.

RESULTSCompared with the end of run-in period, TG, TC, and LDL-C decreased, and HDL-C increased in the treatment group (P < 0.05) after first 3 months of treatment. Compared with 3 months after the first therapy, TG, TC, and LDL-C increased and HDL-C decreased in the treatment group after washout period (P < 0.05). Compared with the end of wash- out period, TC and LDL-C decreased in the treatment group at month 2 after second treatment (P < 0.05); TG, TC, and LDL-C decreased (P < 0.01, P < 0.05), and HDL-C increased (P < 0.05) at month 3 after second treatment. Compared with the control group at month 3 after second treatment, TG, TC, and LDL-C all decreased, and HDL-C increased in the treatment group (all P < 0.05).

CONCLUSIONBC was effective in improving blood lipid level in mild hyperlipidemia patients.

Berberine ; therapeutic use ; Blood Glucose ; analysis ; Body Mass Index ; Capsules ; Humans ; Hyperlipidemias ; drug therapy ; Lipids ; blood

Background Researches demonstrated that the long-term application of glucocorticoids can induce cataract. However, its molecular mechanism is unclear. Objective Present study was to investigate the effects of dexamethasone on the regulation of nuclear factor kappa B( NF-κB)/ inhibitor kappa B alpha( IκBα) line on human lens epithelial cells (LECs) and the LECs apoptosis. Methods Human LECs line(HLE2B3) were cultured and passaged in DMEM containing 20% fetal bovine serum and treated by different concentrations of dexamethasone(0. 01,0. 1,1,10,100 μmol/L) for 24,36 and 48 hours respectively. The LECs cultured in free-serum DMEM without dexamethasone were as blank control group. The expressions of IκBo: in the LECs were examined by reverse transcription polymerase chain reaction ( RT-PCR) and Western blot, and the expressions of NF-κB neucleoprotein in LECs were detected by Western blot after exposure to dexamethasone. The apoptosis rate of LECs was determined by flow cytometer. Results Agarose gel electrophoresis showed that the amplified gene fragment was coincident to designed one. The expressing level of NF-κB neucleoprotein in LECs was significantly lowed with the increase of dexamethasone concentration ( F = 36. 077 , P = 0. 004 ) , and that of IkBo: was evidently ascended ( F = 35. 741 ,P = 0. 002). In the same concentration of dexamethasone group,the expression of NF-κB in LECs showed the considerable alteration in different duration after treated of dexamethasone with the lowest expressing level in 36 hours, and significant differences were found in the expressing level between 24 hours and 36 hours ( P = 0. 002) and between 24 hours and 48 hours (P = 0. 01). The differences of expression of IκBá in LECs appeared the same pattern to NF-κB neucleoprotein. Flow cytometry showed that the apoptosis rate of LECs was obviously enhanced after action of dexamethasone in a dose-dependent manner, showing a significant difference among different groups ( F = 73. 261, P = 0.001). Conclusion It is implied that dexamethasone results in the pathogenesis and development of glucocorticoid cataract by up-regulating the expression of IκBα in LECs and suppressing the activity of NF-κB and herein induce the apoptosis of LECs at concentration-and time-dependent manner. This might be one of cellular and biological mechanisms of glucocorticoid cataract formation.

Objective To type Yersinia pestis isolated from Gansu Province,and to study the trend of diffreent strains in different administrative regions and different years.Methods Totally 193 strains were enrolled in this study,including 9 strains of Ganning Dauricus type,18 strains of Aerjin type,45 strains of Qilian type and 121 strains of Qingzang type.These strains were genotyped by clustered regularly interspaced short palindromic repeats (CRISPR),and genotypes were named according to international standard.Genotyping by CRISPR in different administrative regions and different years of Gansu Province was explored.Results Two clusters (Ca7 and Cb4),including four genotypes (genotypes 7,22,24 and 26) were classified by CRISPR.From the point of view of origin,genotype 24 was the main genotype in Akesai 36.36％ (16/44),Subei 36.17％ (17/47),Yumen 50.00％ (5/10) and Su'nan 38.67％ (29/75);the main genotype of Xiahe and Huining was genotype 26 (4/7);the main genotype of Shandan was genotype 22 (1/1).From the point of view of time,the main genotype of Yersiniapestis in Gansu Province during the years of 1960-1969,1970-1979 and 1980-1989 was genotype 26 [53.33％ (8/15),60.00％ (6/10) and 48.28％ (14/29)];the main genotype was genotype 22 [40.91％ (18/44)] during the years of 1990-1999;and the main genotype was genotype 24 [43.16％ (41/95)] during the years of 2000-2009.Conclusion Four genotypes of Yersiniapestis in Gansu Province are quite different in different administrative regions and different years.

Effect of organic acids on the synthesis of 1,3 propanediol was studied.The adsorption of organic acids from glycerol fermentation liquor by ion-exchange resins was investigated.The results showed that organic acid and 1,3 propanediol production was in negative relationship.The static adsorption showed that ion-exchange resin 005 had the best adsorption abilities of the organic acids in the glycerol fermentation liquor.It was showed that the yield of 1,3propanediol increased by 166% after the extraction of organic acids from glycerol fermentation liquor and the convertion rate increased by 34%.

OBJECTIVETo evaluate the applicability of immune-related response criteria (irRC) in treating non-small cell lung cancer (NSCLC) by Chinese medicine (CM).

METHODSTotally 97 stage III a-IV NSCLC patients were predominantly treated with comprehensive CM. Curative effects were evaluated by three methods such as Response Evaluation Criteria in Solid Tumors (RECIST), Oncologic Curative Effect Evaluation Criteria of Chinese Medicine in Solid Tumor (draft, abbreviated as CM criteria), and irRC. The correspondency and consistency between irRC, RECIST and CM criteria were analyzed and compared. The objectivity of irRC in evaluating curative effect of Chinese medical treatment for NSCLC was assessed.

RESULTSThe correspondency rate of irRC to RECIST was 59. 79% with Kappa value of 0. 379 (U test, P <0. 01). The two criteria had certain correspondence, but with an unsatisfactory consistency. The correspondency rate of irRC to CM criteria rate was 83. 51% with Kappa value of 0.751 (U test, P <0. 01). The two criteria had good correspondence and consistency.

CONCLUSIONSCM criteria had good consistency with CM criteria in evaluating curative effect for Chinese medical treatment of advanced NSCLC. Its results could objectively reflect features and advantages of CM for treating advanced NSCLC.

Asian Continental Ancestry Group ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; immunology ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; immunology ; Medicine, Chinese Traditional ; standards ; Treatment Outcome

Objective To investigate the xenotransplantation model of fetal pig skin precursor tissue and its development after transplantaion. Methods Porcine skin precursor tissue was obtained from the embryo of gestation day 56 (E56), and made into microskin or stamp skin graft. The microskin was transplanted to the dorsal wound in BALB/c nude mice, then covered with human corpse skin. The stamp skin graft was imbedded subcutaneously into the back of nude mice, and microskin was injected subcutaneously into the auricles of nude mice. Their growth and development were observed and they were examined by HE staining at 6th and 12th week after transplantation respectively. Two-sample t test was used to analyze the size of newly grown skin tissue. Results Porcine skin precursor tissue graft in three models above survived and continued growth after transplantation, and growth ability of the dorsal wound transplantation model was significantly stronger than that of the auricle model. Epidermis and hypodermis were detected in newly grown skin tissues. Hair follicles, a few of sebaceous glands, but no sweat glands were observed in auricle model, while many sebaceous glands and sweat glands were observed in the dorsal wound model. Conclusion Transplantation of microskin to dorsal wound is the optimal model of investigating the xenotransplantation of fetal pig skin precursor tissue and its development after transplantion.

Objective To investigate the effect of curcumin pretreatment on endoplasmic reticulum stress induced by global cerebral ischemia-reperfusion (I/R) in rats. Methods One hundred forty-four male SD rats weighing 200-250 g were randomly divided into 4 groups (n = 36 each): sham operation group (group S) ; I/Rgroup; curcumin group (group Cur) and vehicle control group (group VC). Global cerebral I/R was produced by four-vessel occlusion technique in S, I/R, Cur, VC groups. Bilateral vertebral arteries were cauterized. Bilateral common carotid arteries were occluded by clipping for 15 min. Curcumin 200 mg/kg was injected intraperitoneally (IP) at 1 h before cerebral ischemia. Global cerebral ischemia was confirmed by unconsciousness and disappearance of papillary and righting reflex. Animals were sacrificed at 12 h, 1,3 and 7 d of reperfusion. Neuronal apoptosis in hippocampal CA1 region was detected by TUNEL assay. Apoptosis index (AI) was calculated. The expression of glucose regulated protein 78 (GRP78) ,growth arrest and DNA damage inducible gene 153 (GADD153) and caspase-12 protein in hippocampal region was assessed by Western blot analysis. Results Cerebral I/R significantly increased AI and GRP78 and caspase-12 protein expression in hippocampus as compared with group S( P <0.05) . Curcumin pretreatment significantly decreased AI, increased GRP78 protein expression and decreased caspase-12 protein expression as compared with group I/R ( P < 0.05) . There was no significant difference in the GADD153 protein expression among Cur, VC and I/R groups ( P > 0.05) . Conclusion Curcumin pretreatment can significantly reduce global cerebral I/R-induced neuronal apoptosis in hippocampus by increasing GRP78 expression and decreasing easpase-12 expression in hippocampus.

Objective To analyze safety,efficacy and resection methods of video-assisted thoracic surgery(VATS) for the treatment of intralobar pulmonary sequestration(IPS).Methods Data of 17 patients who were diagnosed as IPS and received VATS from December 2006 to September 2011 were retrospectively analyzed.The patients were 7 males and 10 females with the mean age of 40.3 (14-61) years.Diagnosis was confirmed in 9 patients by enhanced CT and unconfirmed in 8 patients.Three ports were used for surgery.After the aberrant artery was confirmed,liner stapler was used in 16 patients to cut it and Hem-o-lok was used in 1 patient because the aberrant artery was about 3 mm in diameter and long enough.If the diameter of the aberrant artery was longer than 10 mm,a stapling device without knife was used to occlude it centrally and a second stapling device was used to cut it peripherally.Wedge resection or lobectomy was performed due to the different conditions.When the lesion was small with linited range in CT image and the lesion was easily distinguished from normal lung tissue during operation,wedge resection was preferred.Results Seventeen patients underwent VATS successfully without any conversion to thoracotomy or any serious complications.Five patients were planned to receive wedge resection and one was converted to lobectomy.Another 12 patients were planned to receive lobectomy and all succeeded.The mean operating time was 128 (80-170)min.The mean blood loss was 80 (5-200) ml.The mean days of chest tube maintained were 4.0 (2-6) days.The mean postoperative hospitalization days were 7.6 (4-11) days.All patients were diagnosed as IPS according to operating in-sight and postoperative pathology.There was no patient suffering from chronic cough,bloody sputum or recurrent pneumonia during the follow-up.Conclusion VATS for the treatment of IPS is safe and feasible.If conditions permit,wedge resection or segmentectomy may be preferred.

Aim To investigate the effects of metformin on phospho-signal transducers and activators of transcription 3(p-STAT3) expression in the spinal dorsal horn in rats with bone cancer pain(BCP).Methods Healthy female SD rats weighing 200~220 g were randomly divided into four groups(n=12 each): Sham+NS, Sham+Metformin, BCP+NS, BCP+Metformin groups.Normal saline or metformin(200 mg·kg-1) was given intraperitoneally once a day at 7~14 day after surgery in four groups.Mechanical withdrawal threshold(MWT) was measured on-1, 3, 5, 7, 10, 12, 14 day after BCP.The L4-6 segments of spinal cords were used to detect the expression of p-STAT3 by Western blot and immunohistochemical staining technique after the behavioral test was completed on day 14.Results Intraperitoneal injection of metformin reversed hyperalgesia and suppressed the expression of p-STAT3 in BCP rats.Conclusion Metformin can attenuate BCP by inhibiting the expression of p-STAT3 in the spinal dorsal horn.