Objective To explore a stable and feasible method for the primary culture of breast cancer-associated fibroblast (CAF), and to offer the theoretical basis for the further studies about the role of CAF in the processing of breast cancer by analyzing the biological property of CAF. Methods CAF was primary cultured with tissue block enzymolytic method. Flow cytometry (FCM) was used to detect the cell cycle. The expression of Vimentin, α-SMA and Ki-67 were detected by cell immunochemistry method. Transwell assay was used to detect the invasion and migration capacity of CAF. Results The CAF was successfullyculturedbyconditionoptimization.ImmunohistochemicalresultsshowedthatVimentin, α-SMA and Ki-67 were highly expressed in CAF.Comparedwithfibroblastsfrombreastcancersidestissues,the proliferating ability and the invasion and migration capacities of CAF were enhanced (number of invasive cells: 79.7 ± 3.5 vs. 66.3 ± 1.5, number of migrated cells: 242.3 ± 3.1 vs. 218.3 ± 2.1, both P<0.05). Conclusion CAF cultured from breast cancer has better proliferating ability and strong invasion and migration capacities.

AIM: To establish a sensitive HPLC method for determining the concentrations of paeonol in human plasma and to evaluate its pharmacokinetic characteristics. METHODS: A single oral dose of 160 mg paeonol capsules was given to 24 Chinese healthy volunteers. Paeonol was separated on a XB-C18 column with tetrahydrofuran-methanol-water-phosphonic acid (6∶60∶34∶0.1, V∶V) as mobile phase. The plasma concentrations of paeonol were determined and its pharmacokinetic parameters were calculated and evaluated using DAS 2.0. RESULTS: The linear range of the paeonol was 10-500 ng/mL and the determination limit was 10 ng/mL. The main pharmacokinetic parameters, as Cmax, tmax, t1/2,AUC0-3, AUC0-∞ after a single dose of paeonol capsules were (116±46)ng/mL,(1.02±0.13) h,(1.03±0.35) h, (174±45) ng/mL, (217±56) ng/mL,respectively. CONCLUSION: The HPLC method for determining paeonol concentration in plasma is rapid, sensitive and suitable for pharmacokinetic studies.

Objective To assess the role of health education in outcomes of diabetes mellitus among high-risk populations.Methods The community physicians who participated this investigation received standardized training,and 307 community residents at high risk of developing diabetes obtained three-month intense health education and nine-month follow-up study.Paired t-test,and Analysis of Variance were used for data analysis.Results After systematic health education,professional level of community physicians was improved.Cognitive level of health knowledge was also significantly improved (5.5 vs 12.6,t=-28.511,P＜0.05).In addition,health knowledge of variant age (F=4.036,P＜0.05),education level (F=15.27,P＜0.05) and occupation (F=9.80,P＜0.05) subgroups was significantly increased.In comparison with baselines,the scores of each age subgroups (F=0.204,P＞0.05) showed no significant differences,although scores of different education level (F=4.71,P＜0.05) and occupation (F=4.87,P＜0.05) subgroups were significantly different.The risk factors of diabetes were effectively controlled.Conclusions Health education should be the key to health management of diabetes,which plays important roles in improving cognitive level of health knowledge among populations at high risk of developing diabetes and reducing the incidence of this condition.

Objective To assess the effects of health management on high-risk diabetic populations.Methods A total of 307 diabetic high-risk adults from 6 communities of Tianjin were recruited by using diabetes risk screening technology.Three-month intensive health management and nine-month follow-up were conducted in this participants.Paired t test for continuous variables and paired contingency table x2 test were used for data analysis.Results Energy intake (1989.8 vs.1766.4 kcal,t =6.84,P ＜0.05),effective exercises (120.4 vs.157.5 kcal,t =-5.00,P ＜ 0.05),body weight (73.0 vs.71.5 kg,t =6.92,P ＜0.05),systolic blood pressure (130.4 vs.124.6 mm Hg (1 mm Hg =0.133 kPa),t =8.36,P ＜0.05),diastolic blood pressure (81.8 vs.78.4 mm Hg,t =7.40,P ＜ 0.05),serum total cholesterol (5.21 vs.5.08 mmol/L,t =2.73,P ＜ 0.05),fasting plasma glucose (6.4 vs.5.8 mmol/L,t =16.37,P ＜ 0.05)and 2 h postprandial blood glucose (7.7 vs.6.9 mmol/L,t =9.67,P ＜ 0.05) were significantly improved after the intervention.Conclusions Community-based health management may provide an effective way to prevent and control the risk factors of diabetes.

Objective To analyze CYP17A1 gene mutation in a patient with 46,XY disordered sex development and to explore the possible influence on the phenotype of the patient.Methods Eight exons of CYP17AI gene in the patient and her parents were amplified and directly sequenced.In order to construct Mini-gene system,PCR fragments containing wildtype and mutant splicing sites were inserted in expression vector,and then transfected into cells.RT-PCR was used to observe the influence of splicing site mutation.Wildtype and aberrant splicing CYP17A1 cDNA expression plasmids were constructed and transfected into cells respectively,and CYP17A1 enzyme activity was tested in vitro.Results Mutation analysis revealed compound heterozygous CYP17A1 mutations,with Y329fs in one allele and a synonymous substitution( c.1263G＞A:GCG＞GCA) in another allele.In vitro analysis showed that the synonymous substitution induced a novel splicing site,which resulted in aberrant splicing of CYP17A1 mRNA and lacked six or seven amino acids after 415 in splicing product.In vitro transfection and enzyme activity experiment showed that the aberrant splicing product abolished the enzyme activity completely.However,this mutation did not completely influence splicing.The patient also had a part of normal splicing product,which was a coincidence to the phenotype of the patient.Conclusion This is the first description of an exonic splicing mutation in CYP17A1 relevant to the 17ot-hydroxylase deficiency phenotype.The functional study of the aberrant splicing variant has been initiated.

Objective To compare the anesthetic effects of interscalene brachial plexus combined with ulnar nerve and axillary brachial plexus block guided by nerve stimulator. Methods Eighty patients belonging to ASA ⅠorⅡ and undergoing replantation of severed palm or wrist were divided randomly into 2 groups, Each group had 40 patients. Nerve stimulator guided nerve block. Patients in groupⅠreceived interscalene brachial plexus combined with ulnar nerve block, and those in groupⅡreceived axillary brachial plexus block. The onset time, hold time, tourniquet tolerance of radial nerve, median nerve and ulnar nerve of two groups was recorded. The phrenic nerve block, Horner′s syndrome and recurrent laryngeal nerve block was compared between two groups. Results The onset time of radial nerve, median nerve and ulnar nerve in group Ⅰwas (5.13 ± 0.76), (7.13 ± 1.04), (3.23 ± 0.62) min , in group Ⅱ was (9.23 ± 1.61), (12.35 ± 1.76), (8.83 ± 1.13) min, and there were significant differences (P<0.05). The excellent rates of sensory block of radial nerve, median nerve and ulnar nerve in group Ⅰ were 90.0%(36/40), 85.0%(34/40), 97.5%(39/40), in group Ⅱ were 72.5%(29/40), 65.0%(26/40), 70.0%(28/40), and there were significant differences (P<0.05). The full rates of motor block of radial nerve, median nerve and ulnar nerve in groupⅠwere 75.0%(30/40), 37.5%(27/40), 80.0%(32/40), in groupⅡ were 47.5%(19/40), 40.0%(16/40), 45.0%(18/40), and there were significant differences (P < 0.05). The tourniquet tolerance rate in group Ⅰwas significantly higher than that in groupⅡ:90.0%(36/40) vs. 62.5%(25/40) , P<0.05. In groupⅠ, phrenic nerve block occurred in 2 patients, and Horner syndrome occurred in 1 patient. None had laryngeal recurrent nerve block in both group. Conclusions The interscalene brachial plexus combined with ulnar nerve block guided by nerve stimulator is more suitable for a long time microsurgery of the palm or wrist, because it takes action faster, has better sensory and motor block effects, improves the rate of tourniquet tolerance without increasing untoward reaction.

OBJECTIVETo assess the safety of repeated invasive prenatal diagnosis primarily due to failed culture of amniotic cells.

METHODSBetween January 2000 and October 2012, 167 cases required repeated invasive prenatal diagnosis among a total of 5304 amniocentesis cases. Clinical outcome and karyotypes were analyzed to calculate the rate of fetal loss.

RESULTSFor the 167 re-sampled cases, the indications have included failed amniocyte culture (121 cases), chromosome mosaicisms (23 cases), failed amniocentesis (21 cases), and request for confirmation (2 cases). No fetal loss has occurred. All samples were cultured successfully. Fourteen cases (8.38%) have been found with an abnormal karyotype. Four mosaic trisomic cases (2 mosaic trisomy 16, 1 mosaic trisomy 20, and 1 mosaic trisomy 8) were verified to be normal.

CONCLUSIONRepeated invasive prenatal diagnosis does not increase the rate of fetal loss. It can be recommended to cases with failed amniocyte culture. Caution should be undertaken when counseling prenatally detected mosaicism trisomies.

Adult ; Amniocentesis ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Karyotyping ; methods ; Middle Aged ; Pregnancy ; Prenatal Diagnosis ; methods ; Trisomy ; Young Adult

OBJECTIVETo explore the effect of N,N-dimethylformamide (DMF) on oxidation or antioxidation status among occupational exposed workers.

METHODSA total of 104 DMF exposed workers and 101 controls were recruited in this study in May 2012. The information of occupational history, age, gender, smoking and alcohol habits of all subjects were collected by questionnaire. DMF concentration in the air of workplace was tested. N-methyl-carbamoylated haemoglobin adduct (NMHb) in blood was chosen as an inner-dose biomarker, which was expressed as 3-methyl-5-isopropylhydantoin(MVH), the degeneration product of NMHb. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) levels were detected to reflect liver function. Serum superoxide dismutase (SOD) activity, glutathione-S-transferase (GST) activity, malondialdehyde (MDA) levels, 3- nitrotyrosine (3-NT) levels were detected to reflect oxidative/antioxidative status.

RESULTSDMF concentration in workplace air was within the range of 3.3-12.4 mg/m(3), which did not exceed our national standard. The MVH level in exposed workers was (19.69 ± 12.52) mg/kg, but was not detectable in control group. The activity of SOD in exposed workers ((125.30 ± 21.23) U/ml) was significantly higher than the control group ((118.35 ± 18.48) U/ml, t = -2.47, P = 0.014). However, the activity of SOD showed different trends with the increasing of MVH level. When MVH ≤ 24 mg/kg, the SOD activity increased with the increasing of MVH level (r = 0.356, P = 0.002). However, when MVH> 24 mg/kg, SOD activity expressed decreasing trend with the increasing of MVH level (r = -0.260, P = 0.150). No significant differences were observed in GST, MDA, 3-NT, ALT, AST levels among the two groups.

CONCLUSIONDMF exposure might have stimulatory effect on antioxidation system of the body under low concentration; within the range of compensatory defense, DMF exposure did not cause obvious lipid and/or protein peroxidative damage.

Adult ; Antioxidants ; metabolism ; Case-Control Studies ; Dimethylformamide ; Female ; Glutathione Transferase ; blood ; Humans ; Male ; Occupational Exposure ; Oxidation-Reduction ; Superoxide Dismutase ; blood ; Workplace ; Young Adult

OBJECTIVE：To optimize the inclusion technology of volatile oil from Ganmao qingre granules. METHODS ： Guided by the concept of “quality by design ”，taking volatile oil inclusion rate and inclusion complex yield as key quality attribute，comprehensive score of above two indexes after weighting as response value ，inclusion temperature ，inclusion time ，the ratio of β-CD to volatile oil as key technology parameters ，Box-Burman response surface design was adopted to establish the design space of key technology parameters and key quality attributes. The design space was optimized and verified by 95％ confidence interval. The stability of inclusion complex was investigated preliminarily. RESULTS ：The optimal design space ，i. e. the optimal technology parameters rang ，included inclusion temperature 35-40 ℃，inclusion time 1.8-2.0 h，the ratio of β-CD to volatile oil 9.5∶1- 10∶1（g/mL）. The results of 3 validation tests showed that the volatile oil inclusion rates were all over 62％，the yields of inclusion complex were all over 75％，and the comprehensive scores were all over 80 point. The results of preliminary stability showed that the inclusion rate of volatile oil ，the yield of inclusion complex and the comprehensive score did not change significantly. The difference in evaluation indicators within 7 days was within 5％. CONCLUSIONS ：The optimized inclusion technology is feasible ， and the obtained inclusion complex is stable.