Aim To study the synergism of silymarin and CH11 on A375-S2 cell apoptosis.Methods Inhibitory effects on A375-S2 cell lines were measured by crystal violet test.Photomicroscopical observation and LDH activity test were used to assess apoptosis.SIRT1,Bcl-2 and Bcl-xL were detected by western blot analysis.Results Treatment with silymarin and CH11 significantly induced apoptosis in A375-S2 cells in a time-dependent manner.A375-S2 cells incubated with silymarin and CH11 for 12 h showed marked apoptotic changes including condensed chromatin,nuclear fragmentation and apoptotic bodies.Expressions of anti-apoptotic Bcl-2 and Bcl-xL(Bcl-2 family member) were decreased simultaneously.Expressions of apoptotic Bax and cytochrom C were increased.Consequently,Caspase-3,downstream of mitochondria signal pathway, more significantly expressed after A375-S2 cells treatment with silymarin and CH11 for 12 h.Conclusion Silymarin and CH11 synergistically induced A375-S2 cells apoptosis.

Laboratory diagnostics is one of the most fast developing medical sciences.But the teaching quality falls behind due to the traditional education model.Based on the concept of ‘three-stage fusion' of the subject and ‘five abilities' of the training objects,‘three-highlight & stereo' teaching style was established and put through the course,which means to emphasize function,practice,effect in optimizing teaching content and methods,and construct stereo platform including information system,online course,laboratory equipments,research activities and teacher training program.The results showed a significant improvement of the students' knowledge mastering and utilizing capability.And the teaching situation was well re-created.Furthermore,the teaching team was much more powerful than ever.It suggests that ‘three-highlight and stereo' teaching style is a successful explore and practice of teaching reform of laboratory diagnostics on adapting for internationalization of education tendency.

Objective To investigate the role of mitogen-activated protein kinase(MAPK)signaling pathway regulating synthesis function of steroid hormone of porcine ovarian granulose cells.Methods Porcine ovarian granulose cells were cultured in medium added with dimethyl sulfoxide(DMSO)at concentration of 50μmol/L or with MAPK inhibitor PD98059 (PD) for 48 hours.Then,granulose cells in DMSO medium were added with activator of adenylate cyclase(20 μmoL/L)or blank agent for next 48 hours incubation,which were defined as observation group 1 and control group 1,similarly,granulose cells in PD medium were also added with activator of adenylate cyclase(20 μmol/L)or blank agent for next 48 hours,which were defined as observation group 2 and control group 2.The level of estradiol(E_2),progesterone (P)and testosterone (T) were detected by chemoluminescence.The mRNA expression of 17alpha-hydroxylase/17,20-lyase (CYP17) and aromatase P450 (P450 arom) were assessed by RT-PCR.Results (1)Hormone expression:the level of T,P and E_2 were(29.5±2.5)nmol/L,(80±5)nmol/L,(49±4) pmol/L in control group 1 and(42.3±3.4)nmol/L,(170±15)nmol/L,(75±6)pmol/L in control group 2,which showed significant difference(P<0.05).In the mean time,the level of T, P and E_2 were (106.2±7.6)nmol/L,(210±16)nmol/L,(130±11)pmol/L in observation group 2 and(47.2±3.5)nmol/L,(130±6)nmol/L,(81±6)pmol/L in observation group 1,which also reach statistical difference(P<0.05).(2)The expression of enzyme:the expression of CYP17 mRNA was inereased by 50% and P450 arom mRNA was decreased by 20% between control group 1 and 2. However, the mRNA expression of P450 arom and CYPI7 were upregulated remarkably, especially, the expression of CYP17 mRNA were increased by 125%. Conclusion MAPK signaling pathway plays an inhibitory role in regulating synthesis of steroid hormone of ovarian granulose cell.

Objective: To clone human carboxypeptidase A1 and its active center gene as well as construct recombinant vector for expression before analysis their activities. Methods: CPA1 and CPAlactive center gene were amplified by RT-PCR from pancreas tissue, and then sequencing was carried out. The correct target genes were cloned into prokaryotic vector pGEX-4T-1 and transformed into E. coli BL21 before sequence analysis. After induced by IPTG, gene products were analyzed by SDS-PAGE. Target expressed proteins were denaturated, renaturated, purified and evaluated through MTT and agar colony form test. Results: Human carboxypeptidase Al and its active center gene were cloned successfully. New expected protein band of Mr 66,000 and 46,000 appeared on SDS-PAGE after inducement. Both of the expressed proteins have catalytic activity in vitro, but the activity of the latter is inferior when applied to tumor cells. Conclusions: Human carboxypeptidase A1 and its active center gene were cloned successfully. Their prokaryotic expression products were obtained too. The expressed proteins have catalytic activity in. vitro. A new prosperous beginning of further improvement for CPA1 therapy system has been established based on CPA1 and its active center gene in terms of ADEPT against prostate cancer to clinical application.

OBJECTIVE To enhance the positive rate of the blood culture in order to make pathogenic diagnosis actually and quickly and conducting usage of antimicrobial agents in clinic.To value the clinical applied circumstance of BacT/Alert 3D automated blood culture system.METHODS The 3728 blood cultures were detected by BacT/Alert 3D automated blood culture system,and bacterial susceptibility test was conducted on all isolates using Kirby-Bauer methods with CLSI standards.To statistically analyze the examined time,the positive rate and variety and drug resistance for all kinds of pathogens.RESULTS The blood culture positive rate was 6.0% in the 3728 blood cultures with 222 strains of bacteria.The false positive rate was 0.2% and the false negative rate was 0.4%.The positive rate of blood culture was 0.89% in 12 hours,2.01% in 18 hours,and 3.51% in 24 hours.Among all the 222 isolates,29.7% were Enterobacteriaceae,the drug resistant rates to amikacin,ceftazidime,ciprofloxacin,and imipenem were 13.6%,36.3%,42.4%,and 3.0%,respectively;20.3% were Staphylococcus,the drug resistant rates to erythromycin,levofloxacin,cefoxitin,and vancomycin were 75.6%,33.3%,68.9%,and 0,respectively;11.7% were Enterococcus,the drug resistant rates to errythromycin,levofloxacin,fosfomycin,and vancomycin were 96.2%,80.8%,23.1%,and 0,respectively;11.3% were non-fermented bacilli,the drug resistant rates to amikacin,ceftazidime,ciprofloxacin,and imipenem were 32.0%,52.0%,32.0%,and 32.0%,respectively;12.6% were fungi.CONCLUSIONS The pathogens in the blood specimens are more wider in distribution and more higher in drug resistance rates than before.BacT/Alert 3D automated blood culture system can be an important tool for the blood culture,it can provide the diagnostic help quickly for the clinic and increase the positive rates in blood culture.It can not only shorten the check-up time,but also be more quick and more exact.

This study was aimed to establish an analysis method of amentotaxus biflavone distribution in rats' tissues, including the heart, liver, spleen, lungs, kidney, brain, stomach, large intestine and small intestine, in order to investigate its distribution characteristics in tissues after rats gavaged with amentotaxus biflavone. HPLC-UV was employed to determine contents of amentotaxus biflavone in rats' tissues. The intragastric administration dose of amentotaxus biflavone was 500 mg·kg-1. Rats were sacrificed by cervical dislocation 10 min, 0.5, 1, 2, 4, 8, 12 h after intragastric administration. Tissues of the heart, liver, spleen, lungs, kidney, stomach, large intestine, small intestine and brain were removed and dissected immediately. Distribution of amentotaxus biflavone in each tissue was determined after processing. The results showed that tissue in different range had a good linear range. The lowest detection limit was 0.125 ng. The RSD of intra and inter-day was less than 10%. The absolute recovery rate of amentotaxus biflavone in each tissue was between 75.07% and 89.80%. The relative recovery rate was between 92.00% and 107.00%. Amentotaxus biflavone in each tissue was stable in different tissues in refrigerator of -20° C for 15 days. It was concluded that there were relatively large concentration differences of amentotaxus biflavone in different tissues. After intragastric administration, amentotaxus biflavone was mainly distributed in the stomach, large intestine, small intestine, liver and kidney, and then the heart, lungs and spleen. It also distributed in brain tissues through the blood-brain barrier.

Objective The aim of this study was to observe the anesthetic effect of xylazine hydrochloride Injection on Beagle dogs.Methods Anaesthetizing 30 healthy Beagles with xylazine hydrochloride injection, some physiological indexes of the dogs, such as body temperature (T), respiration (R), heart rate (P), and blood pressure (systolic pressure SBP, diastolic pressure DBP), were monitored.Results After using xylazine hydrochloride injection, the body temperature, respiration, heart rate, blood pressure, and peripheral vascular resistance were decreased in the Beagles.Conclusion Xylazine hydrochloride injection can keep a stable induction period, and has advantages including powerful and quick effect, simple method and operation, safe and insignificant toxicity and side effects,and has a better effect of anaesthesia.

OBJECTIVE: To observe the therapeutic effects of Jinqi Jiangtang Tablet (JQJTT), a commonly prescribed recipe for type 2 diabetes in traditional Chinese medicine, on obese women with polycystic ovary syndrome (PCOS). METHODS: Twenty-four obese women with PCOS were prescribed JQJTT for 3 months and the changes of carbohydrate metabolism and biochemical indicators, including menstrual cycle, body mass index (BMI), the level of blood sex hormone, area under curve (AUC) of glucose and insulin during oral glucose tolerance test (OGTT), and testosterone responses during human chorionic gonadotrophin stimulation ovary test were observed. RESULTS: After treatment, there were significantly reductions in BMI (P<0.05), serum glucose level at 30 min during OGTT (P<0.05) and testosterone levels at eighteen hours during human chorionic gonadotrophin stimulation ovary test (P<0.05). In addition, nine patients had improved cycles among sixteen completed patients, and one patient with clomifene resistance got pregnant with response to clomifene treatment. CONCLUSION: The beneficial effects of JQJTT may result from improvement of carbohydrate metabolism and reduction in androgen biosynthesis in women with PCOS.

Objective: To observe the haemodynamic changes within ovarian stromal blood flow in patients with polycystic ovary syndrome(PCOS). Methods: 41 patients with PCOS and 10 ovulatory women underwent transrectal ultrasound associated with Doppler flow measurement of the artery within ovarian stroma during the early follicular phase (days 2-5 of the menstrual cycle) or any day in patients with amenorrhea . Results:The peculiar vascular pattern wich one prominent blood vessel showed passing through the ovarian stroma from ovary gate. This sign was observed in all ovaries in patients with PCOS.There was no significiant difference in Vmax、PI、RI between left and right ovary.The Vmax,PI,RI of the right ovarian stromal artery were (18.99?3.12)cm/s, 0.74?0.12, 0.50?0.03 respectively. The indices of the left ovarian stromal artery were (18.76?3.23)cm/s, 0.75?0.11, 0.49?0.04 respectively. Conclusion: Vascularization of the ovarian stroma is prominent in patients with PCOS than in the control. The decreased vascular resistance to ovarian stromal perfusion in patients with PCOS may lead to perfusion increasment. TRS can be used as a non-invasive, convenient, and sensitive method for assessing the blood flow within the ovarian stroma.

As a new type of intercellular signaling rector, extracellular vesicles (EV) are involved in almost the whole process of tumorigenesis, progression, metastasis, and drug resistance. Therefore, EV have become the ideal biomarker candidates and research hotspots for cancer diagnosis and treatment. However, EV tumor biomarker research mainly focused on RNA and protein, and a small part of the research focused on lipids at the early stage. EV DNA has received little attention and its diagnostic value has gradually been recognized in recent years. Study on the biological characteristics and function of EV DNA may highlight its potential in tumor diagnosis.