Objective To evaluate the effect of puerarin on melanogenesis in melanocytes,and to explore its possible mechanisms.Methods Third-to fifth-passage melanocytes isolated from human foreskin were treated with different concentrations (1,5,10,20,40,80 and 160 μmol/L) of puerarin for 24 hours,with those receiving no treatment as the normal control group.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of melanocytes,a sodium hydroxide solubilization method was used to measure melanin content,and reverse transcription PCR (RT-PCR) and Western blot analysis were performed to quantify the mRNA and protein expressions of microphthalmia-associated transcription factor (MITF),tyrosinase (TYR) and tyrosinase-related protein-1 (TRP-1) respectively.Results There were no significant differences in the proliferative activity of melanocytes between the puerarin (1-40 μmol/L) groups and normal control group (P ＞ 0.05),and 40 μmol/L was chosen as the concentration of puerarin for subsequent experiments.Compared with the normal control group,the 40-μmol/L puerarin group showed increased melanin content as well as mRNA and protein expressions of MITF,TYR and TRP-1 (all P ＜ 0.05).Concretely speaking,the protein expressions of MITF,TYR and TRP-1 in the 40-μmol/L puerarin group were increased by 8.69％,10.28％ and 10.58％ compared with the normal control group respectively (all P ＜ 0.05),and their mRNA expressions were 2.48,1.91 and 1.63 times higher in the 40-μmol/L puerarin group than in the normal control group respectively (all P ＜ 0.05).Conclusion Puerarin can increase the mRNA and protein expressions of MITF,TYR and TRP-1,and promote melanogenesis in melanocytes.

Objective To study the changes of hematological indices of blood donors after 400ml whole-blood donation.Methods Blood specimens of 123 healthy donors were collected before donation,1 day,7 days,1 month,and 3 months after donation.The dynamic changes of WBC,RBC,hemoglobin,hematocrit,Plt,plasma total protein,albumin and blood serum iron were observed.Results After donation,the changes of WBC,Plt,total protein of blood plasma and albumin showed no significant change.The levels of RBC,hemoglobin,hematocrit and blood serum iron returned to normal after 3 months.Conclusion For anyone who meets the health standard of blood donors,to donate 400ml whole blood at 3-month interval is harmless.

Objective To evaluate the potential utility of temporal temperature gradient gel electrophoresis (TTGE) in detecting homoplasmic and heteroplasmic mtDNA mutations in patients with cancers. Methods The entire mtDNA of tumor and corresponding normal tissues from total 117 patients were amplified by PCR, and the somatic mutations were screened by TTGE. DNA fragments showing different band patterns in normal and tumor tissues in TTGE were sequened to identify the exact mutations. Results 160 fragments harbouring mtDNA mutations were found by TTGE in total 3 654 fragments. Compared with the direct squencing results, the sensitivity and specificity of TTGE in mutation detection were 96.27% and 94.05% respectively. Conclusion TTGE is an effective method for screening acquired homoplasmic mitochondrial gene mutation and its different percentage.

Objective To investigate the relevant factors of neonatal ventilator-associated pneumonia(VAP),and to provide a theoretical basis of prevention and treatment.Methods Retrospective analyed the clinical data of 145 critically ill neonates,who were treated with mechanical ventilator from Jan 2006 to Dec 2009 in the Third People′s Hospital of Wenzhou City,NICU.According to whether the neonates were occurred VAP,they were divided into two groups:VAP group(52 cases) and without VAP group(93 cases).Results Fifty-two out of the 145 neonates developed VAP.The incidence of VAP was 35.86%,the main relevent factors were the gestational ages,birth weights,the duration of mechanical ventilation and the times of intubation.There was significant difference between the two groups (P<0.05).The main pathogens were opportunistic bacteria,and mostly were G-bacilli.Conclusion The incidence of VAP has a close conclusion with the gestational ages,the birth weights,the duration of mechanical ventilation and the times of intubation.Regulate the use of breathing machine,strengthen aseptic operation,and select effective antibiotic can control the occurrence and development of VAP.

This article was aimed to study effects and mechanisms of Gymnema sylvestre on protein kinase B (PKB) and its phosphorylation in adipose tissues of KKAy mice which were mainly characterized by insulin resistance (IR). A total of 18 KKAy mice were randomly divided into the diabetes model (DM) group and Gymnema sylvestre (GS) group according to body weight levels. And 9 normal C57BL/6J mice were used as the normal control (NC) group. Intragastric administration of medication was given to mice for 8 weeks. At the end of the experiment, all animals were tested for fasting plasma glucose (FPG) and fasting insulin level (Fins) for evaluation of insulin sensitivity index (ISI). Expressions of phosphoinositide-dependent kinase-1 (PDK1), PKB, P-PKB (Ser473), P-PKB (Thr 308) in adi-pose tissues of epididymis were determined. The expression of phosphatase and tensin homolog (PTEN) mRNA was also determined. The results showed that compared with the DM group, the GS group showed lower FPG and Fins, higher ISI. The expression of P-PKB (Ser473) phosphorylation and P-PKB (Thr 308) were increased, and the PDK1 and PTEN mRNA were decreased. It was concluded that GS can improve insulin sensitivity of KKAy mice through activating PKB by up-regulate the expression of P- PKB (Ser473) and its phosphorylation ratio and P- PKB (Thr 308) in adipose tissues.

Objective To evaluate ultrasonic measurement of fetal thigh soft tissue thickness in the estimation of fetal weight . Methods Fetal biparietal (BPD), head cirumference (HC), abdominal cirumference (AC) , femur length (FL) and fetal thigh soft tissue thickness (TSTT) were measured by ultrasonography in 196 pregnant women during five days before dilivery and the relation with neonatal birth weight were analyzed.Results There was significantly correlation between TSTT and neonatal weight (r=0.865) , and it was better to estimate fetal weight with TSTT than with the other parameters. The prospective study showed that the predictive value of fetal TSTT was 80 1%.Conclusions The ultrasonic measurement of fetal TSTT is a simple and valuable index in estimation of fetal weight. [

Objective To establish a sensitive and specific liquid chromatography method for the determination of loganin and morroniside in rat serum containing Liuwei Dihuang Formula(LDF).Methods Loganin and morroniside were extracted from the rat serum using SPE column,then separated on a C_(18)column.A HPLC method for determining serum concentration of mor- roniside and loganin in rats before and after administration LDF was established.Results A good linear calibration curve of lo- ganin and morroniside was obtained.The recovery of the method was in the range of 95 %～105 %.The extracted recovery was more than 75 % ,and within-day and intra-day precisions were less than 12 %.Conclusion This method is sensitive, simple and rapid for monitoring of drug level in the study on pharmacokinetics and different combination of LDF.

OBJECTIVE:To evaluate the quality of anticoagulant therapy in patients who took warfarin and provide data sup-porting for strengthening the management of these patients. METHODS:A retrospective analysis of related clinical data of 214 pa-tients who meet the inclusion criteria was performed. RESULTS:The average time for patients took warfarin was (321.64 ± 189.50)d,average times for tested INR was(12.01±7.03)times in clinic;when anticoagulant therapy,INR<2.0 accounted for 51.96%,2.0≤INR≤3.0 accounted for 39.13%,and INR>3.0 accounted for 8.91%;patients with target INR≤2.0 accounted for 45.33%,target INR=2.5 accounted for 38.32%,and target INR≥3.0 accounted for 0.93%;the average TTR was (50.80 ± 22.32)%;and there was no statistical significance in the TTR of different ages and diseases(P>0.05). CONCLUSIONS:The anti-coagulant therapy in some patients who took warfarin shows poor quality,it needs strengthening the quality management to make it safe and effective.

Objective: To explore influence of nursing health education on morning blood pressure surge (MBPS) fluctuations in aged patients with hypertension.Methods: A total of 100 outpatients and inpatients with non-dipper hypertension, who haven't been treated, were selected from Apr 2011 to Sep 2015.According to random number table method, patients were randomly and equally divided into routine treatment group and health education group (received nursing health education based on routine treatment group), both groups were treated for four weeks.MBPS, trough/peak(T/P) ratio and smoothness index (SI) were measured and compared between two groups after treatment.Results: Compared with routine treatment group after four-week treatment, there were significant reductions in systolic blood pressure (SBP) morning surge[(46.95±5.17) mmHg vs.(42.21±5.19) mmHg]and diastolic blood pressure (DBP) morning surge[(16.31±3.23) mmHg vs.(14.21±3.58) mmHg], and significant rise in SBP T/P[(59.22±6.65)% vs.(70.11±6.41)%], DBP T/P[(50.91±5.04)% vs.(56.97±5.12)%], SBP SI[(1.19±0.13) vs.(1.42±0.12)]and DBP SI[(1.11±0.09) vs.(1.29±0.08)]in health education group, P<0.05 all.Conclusion: Health education can effectively improve blood pressure fluctuations and may likely improve prognosis in patients with non-dipper hypertension, which is worth extending.

OBJECTIVE:To explore the optimal steaming time of Carapax trionycis during cleansing period,and to optimize and improve production technology of Carapax trionycis recorded by current Chinese Pharmacopoeia. METHODS:The mechanical processing replaced the artificial processing method in Chinese Pharmacopoeia. The content of protein and the appearance of Cara-pax trionycis were investigated after steaming for 30,60,90,120,180,240 min during cleansing period. The extract,decoction, ash content,appearance and property of Carapax trionycis decoction piece processed with vinegar were also investigated after cleansed Carapax trionycis decoction piece was processed by sand scalding and vinegar quenching method. RESULTS:The differ-ent steaming time obtained different quality of cleansed Carapax trionycis decoction piece and Carapax trionycis decoction piece pro-cessed with vinegar. Compared with decoction piece steamed for other duration,when the steaming time was 90 min,the content of protein in cleansed Carapax trionycis decoction piece was higher(31.16%),and its appearance was up to the requirement. Cara-pax trionycis decoction piece processed with vinegar had higher contents of extract and decoction(9.13%,11.39%)and lower con-tent of ash(66.29%),and its appearance was up to the requirement. CONCLUSIONS:Different steaming time have certain effect on the quality of cleansed Carapax trionycis and Carapax trionycis processed with vinegar,the optimal steaming time of Carapax tri-onycis is about 90 min during cleansing. The mechanical processing method maybe replace the artificial processing on Carapax tri-onycis for improving its production efficiency.