BACKGROUND: It is difficult to in vitro isolate and culture the ovarian surface epithelium with high purity, strong vitality and stable biological characteristics. Tissue adherence and enzymatic digestion are commonly used for primary culture, but there are certain problems in cel col ection, cel viability and cel purity. OBJECTIVE: To establish a method for primary isolating, culturing and identifying human ovarian epithelium. METHODS: The ovarian surface epithelium was obtained with cel brush innovatively, and then the cells were isolated and purified with erythrocyte spal ation and differential adherence. The epidermal growth factor was added into the serum-free Dulbecco’s modified Eagle’s medium-F12 medium for cel culture. The cel morphology was observed under inverted microscope, and hematoxylin-eosin staining and immunocytochemical staining were used to identify the cells, then the growth curve was draw. RESULTS AND CONCLUSION: The ovarian surface epithelium became adherent after cultured for 24 hours, and reached fusion basical y after cultured for 6-12 days. The cells were polygonal or flat with strong transparency and refraction. The morphological characteristics of the cells were in line with those of the normal epithelial cells, and almost al the isolated cells could express the epithelial cells surface marker CK19. The cells could be passaged for 6-8 generations with wel growth and the cel growth curve was in “S” shape. The purity of the cells was more than 95%. The results suggest that cel brush method is simple to operate and can obtain a large amount of ovarian surface epithelium rapidly. The purity of the isolated cells can reach to 95% after treated with erythrocyte spal ation and differential adherence method and the cells are in stable growth.

Objective To evaluate the status of iodine deficiency disorders(IDD) in Huai'an City in 2011 and to provide a scientific basis for adopting target prevention strategies timely and adjusting the intervention policy scientifically.Methods According to The National Project of Surveillance on IDD,probability sampling method (PPS) was used to select 30 counties in Huai'an City,and one primary school was selected randomly from each county,then 40 children aged 8 to 10 were randomly selected in each school.Children's thyroids were examined by palpation and the iodine in salt samples collected in their home was tested by direct titration.At the same time,urine samples from 12 children of the 40 selected children were collected and urinary iodine was tested by As(Ⅲ)-Ce4+ catalytic spectrophotometry.Questionnaire survey of IDD was conducted among 30 students of grade 5 and 5 housewives around that school.Results A total of 1200 children aged 8 to 10 were examined,and the goiter rate was 1.3％(15/1200).All the goiters were level Ⅰ.The goiters of each age group was 0.8％(3/390),1.8％(7/390) and 1.3％(5/420),respectively.The 360 urine samples were collected from children aged 8-10,and the median of urinary iodine was 171.5 μg/L with the proportions of ＜ 20 μg/L,＜ 50 μg/L and ＜ 100 μg/L were 0(0/360),1.1％ (4/360) and 5.6％ (20/360),respectively.A total of 1200 salt samples were tested in children's family.The coverage rate of iodized salt was 99.4％ (1193/1200) and the qualified rate of iodized salt was 97.7％ (1165/1193).The consuming rate of qualified iodized salt was 97.1％(1165/1200),and non iodized salt rate was 0.6％(7/1200).The knowledge rates of IDD were 82.7％ (2234/2700) and 69.8％ (314/450) in 900 students and 150 housewives,respectively.Conclusions In Huai'an City,except the knowledge rate of IDD,the goiter rate and urinary iodine of children aged 8-10,iodized salt consumption maintain at the stage of the target level.In addition to strengthen the monitoring of iodized salt and to supply qualified iodized salt,it should also increase the focus on health education propaganda in the future.

Objective To explore the diagnosis and management of functional pancreatic endocrine tumor.Methods Clinical data of 19 cases of functional pancreatic endocrine tumor were retrospectively analyzed.Results 15 cases of insulinoma,2 cases of gastrinoma and 2 cases of glucagonoma were qualitatively diagnosed.The positive rate of preoperative diagosis for type B ultrasonic inspection,CT,MRI,EUS,selective portovenous sampling and intraoperative type B ultrasonic inspection was 15.8％ (3/19),67.5％ (10/16),71.4％ (5/7),87.5％ (7/8),100％(2/2) and 85.7％(6/7) respectively.Of the total 19 cases,7 cases underwent open surgery,11 cases unde rwent laparoscopic surgery,and one case didn't undergo any surgery as liver metastasis had occurred when glucagonoma was diagnosed.The operation methods included tumor enucleation (n=13),distal pancreatic resection (n=3),distal pancreatic resection plus splenectomy (n=1),and pancreatic head resection with duodenum preserved (n=1).Conclusions The measurement of serum insulin,gastrin and glucagon is the main basis for qualitative diagonosis of pancreatic endocrine tumor.Two stage spinal CT thin scanning is the main method for tumor location.Intraoperative type B ultrasonic inspection is the supplement to preoperative location.Tumor enucleation is the main choice of treatment.

Objective:To observe the therapeutic effects of Tongyushu capsules on dysmenorrhea. Methods:With the intravenous injection of oxytocin in rats, the influence of Tongyushu capsules on rat uterine smooth muscle was observed in vivo. With the intraper-itoneal injection of oxytocin to replicate the mouse model of dysmenorrhea, the effect of Tongyushu capsules on dysmenorrhea was ob-served. Using acetic acid writhing method, the effect of relieving pain Tongyushu capsules in mice was observed. By the method of ear swelling induced by xylene in mice, the anti-inflammatory effect of Tongyushu capsules was observed. By the method of ice water bath and subcutaneous injection of adrenaline, the influence of Tongyushu capsules on the acute blood stasis model in rats was observed. Results:Compared with those of the model group, the contraction of uterine smooth muscle of the groups treated with Tongyushu cap-sules at various dosage was decreased obviously (P<0. 05), the number of body torsion reaction induced by oxytocin and acetic acid in mice was obviously decreased in 15 min (P<0. 05), the degree of ear swelling in mice treated with Tongyushu capsules was signifi-cantly reduced (P<0. 05), and the whole blood viscosity, plasma viscosity, hematokrit, erythrocyte sedimentation rate and erythro-cyte electrophoresis time were significantly decreased (P<0. 05). Conclusion:Tongyushu capsules have obvious effects of the inhibi-tion of excessive contraction of uterine smooth muscle, dysmenorrhea alleviation, anti-inflammation and relieving pain, which show good therapeutic effect on dysmenorrhea.

Objective To investigate the carbapenem-resistant mechanism of one strain of Klebsiella pneumoniae( KPN) and one strain of Pseudomonas aeruginosa( PAE) .Methods The identity of the isolates was confirmed by using MALDI-TOF mass spectrometry, 16S rDNA and special gene amplification .The minimum inhibitory concentrations (MICs) of the antimicrobial agents were determined by VITEK 2 Compact System .CarbaNP Antimicrobial susceptibility testing , plasmid extraction, electroporation experiment , PCR amplification, and cloning and sequencing were carried out to analyze the en-coding gene of β-lactamases.Results The types of β-lactamases of the KPN were blaKPC-2 and blaSHV, confirmed by se-quencing of the PCR products , and that of the PAE were blaKPC-2 .Only blaKPC-2 was displayed in both transformants .All of the results of CarbaNP were type A .Conclusion Both strains of KPN and PAE resisting to carbapenem produce a plasmid-mediated carbapenemase blaKPC-2 , which belongs to Bush group 2f, class A β-lactamase.The extended-spectrum β-lacta-mases gene encoding blaSHV of the KPN might be located in the chromosome or not in the plasmid carrying with blaKPC-2 .

Objective To investigate treatment effects of lentivirus mediated RhoA short hairpin RNA(shRNA) on xenograft tumor of ovarian cancer in nude mice in vivo and the underlying mechanism.Methods Human ovarian cancer cell line HO8910 were inoculated to establish subcutaneous xenograft model of human ovarian cancer.Tumor-bearing nude mice were assigned randomizely to three groups:LentiRhoA-sh group,Lenti-negative control (NC) group and phosphate buffered saline (PBS) group.lentivirus mediated RhoA shRNA,negative control lentivirus and PBS were respectively injected in the three groups.Effects of treatment were observed by tumor growth curve,tumor volume,tumor weight,and tumor inhibition rate.Xenograft tissues and liver,spleen,lung,and renal tissues were examined by hematoxylin and eosin (HE) staining or were detected by streptavidin-perosidase (SP)immunochemical method.The changes of RhoA gene expression in xenograft tissues after lentivirus mediated RhoA shRNA treated were also detected by real-time qPCR,immunochemistry and Western blot assay.Cell apoptosis in xenograft tissues were examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) method and apoptotic index (AI) were counted.Results Compared with Lenti-NC group and PBS group,the growth speed of xenograft in Lenti-RhoA-sh group delayed significantly after injection 9 days(P ＜ 0.01).Tumor volume (338 ± 114) mm3 decreased significantly in the Lenti-RhoA-sh group when compared with those in Lenti-NC group(1190 ± 332)mm3 and PBS group (1101 ± 396) mm3 (P ＜ 0.01).Tumor weight (0.23±0.11)g decreased significantly in the Lenti-RhoA-sh group when compared with Lenti-NC group (0.79 ± 0.19)g and PBS group (0.74 ± 0.17)g (P ＜ 0.01).Real-time qPCR result shown that the expression of RhoA mRNA (0.30 ± 0.05) decreased significantly in the Lenti-RhoA-sh group compared with Lenti-NC group (0.95 ±0.06) and PBS group(1.00 ±0.11 ; P ＜0.01).Western blot result showed that the expression level of RhoA protein decreased significantly in the Lenti-RhoA-sh group (0.14 ± 0.06) compared with those in Lenti-NC group(0.78 ± 0.14) and PBS group (0.75 ± 0.13;P ＜ 0.01).TUNEL staining displayed that AI significantly increased in the Lenti-RhoA-sh group (20.9 ± 3.4) ％ compared with those in Lenti-NC group(5.2 ±±2.0)％ and PBS group(6.0 ±2.1)％ (P ＜0.01).Conclusion Lentivirus mediated RhoA shRNA may be effectively down-regulate of the expression of RhoA,inhibit the growth of subcutaneous xenograft tumor of ovarian cancer in nude mice by increasing the cell apoptosis.

Cholestasis ; etiology ; Gestational Age ; Humans ; Hydrops Fetalis ; physiopathology ; Infant, Newborn ; Liver Function Tests ; Male ; Tachycardia, Paroxysmal ; complications ; embryology ; physiopathology ; Tachycardia, Supraventricular ; complications ; embryology ; physiopathology

To screen the illegal substances in fishery inputs,we established the database including the precursor and the daughter ions for these possible components by the quadrupole/orbit-trap mass spectrometer,and the retention time of each drug on the same chromatographic column.And then,the extracted and diluted samples were analyzed and the components in the real samples were identified under the same conditions.Chromatographic analysis was performed on an Accucore RP-MS column (100 mm × 2.1 mm,2.6 μm) using gradient elution with 0.1％ formic acid in water and 0.1％ formic acid in acetonitrile as mobile phase.Elutes were ionized through heatable electrospray ionization (HESI) in both positive and negative mode simultaneously.Data acquisition was conducted by Full-scan ddMS2 (TopN) mode,in which the full mass profile for a continuous precursor ion injection and the fragments of each high abundant precursor of targeted were acquired with excellent time and mass resolution.Screening was carried out through comparison of the information of real samples with that of standards in the database,which were processed by software (Tracefinder).The Quantification of each component was analyzed based on the precursor ion chromatography acquired by orbit-trap mass spectroscopy,which showed a good linearity between 0.01-1 μg/mL,with R＞0.98.The method was validated by checking its minimum screening concentration (0.5 mg/L for drugs and 5 mg/L for feedstuffs) and evaluating the recovery after addition of the standard mixture in real samples (＞50％,under the addition of 10 and 100 mg/kg).The results for 68 practical samples demonstrated the effective performance of this method for screening with high-throughput,rapidness and acceptable minimum screening concentration and accuracy,in which 15 of 29 fishery drug samples were screened out for positive components that were not indicated in their labels.

Objective To retrospectively analyze the effect of abdominal acupuncture for the casino workers with sleep disorder in chronic fatigue syndrome and analyze the correlation between them.Method 65 patients were all from the Acupuncture Department of Health Center of the Black Sand, Macao Health Bureau, 2011 October to 2013 January, who were diagnosed with sleep disorder in chronic fatigue syndrome and in accordance with the inclusion criteria. They were received abdominal acupuncture treatment for 4 weeks. Pittsburgh sleep quality index(PSQI) and Fatigue Assessment Instrument(FAI)were adopted to evaluate the effect before treatment 、after treatment and one month follow-up. Multiple regression analysis model was used to analyze the correlation of each factor of FAI and PSQI. Results Compared with before treatment (13.78± 3.23), the total score of PSQI after treatment(8.66 ± 4.26)and follow-up(8.26 ± 4.66) were significant decreased(P<0.05);the score of 6 dimensions(sleep quality, Time to fall asleep, sleep time, sleep efficiency, sleep disorders daytime function) after treatment and follow-up were significant decreased(P<0.05),compared with those before treatment;the FAI score after treatment (92.60±14.64) and follow-up(90.75±14.75) were significant lower(P<0.05)than those before treatment (147.55±7.03) .Multiple regression analysis showed that 4 dimensions of PSQI (sleep quality, sleep time, sleep disorders, daytime function)had close relationship with FAI score(r=0.373~0.702, P<0.05). Conclusion Abdominal acupuncture can improve the casino workers' sleep disorder in chronic fatigue syndrome and is worth of further study.

AIM: To investigate serum lipid and the expression of SR-B1 in the livers of diabetic mice. METHODS: Ten normal diet, female C57BL/6J mice, fifteen high fat and sugar diet female C57BL/6J mice, five fed 8 weeks and ten fed 16 weeks were used in the experiment. Serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C), fasting blood glucose (FBG), insulin (INS) and the expression of SR-B1 in the livers were measured. RESULTS: 1. In the high fat and sugar diet mice, serum TC and FBG at 16 weeks were significantly higher than that in normal diet mice (P