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 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.

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.

AIM: To study the pharmacodynamics of Yajiao Hadun San(Vitex trifolia;streptocaulon griffibhii;Asparagus pseudofilicinus;Inula cappa;Benincasa hispida;Tacca esquirolli). METHODS: The pyrexia model rat induced with active yeast,the mice's auricular swelling test caused by croton oil;the mice's toe swelling test caused by albumen;the mice's twisting body test caused by the irritation of acetic acid and the mice's carbon clearance test were used to determine antipyretic,analgesic,anti-inflammation effect and immunity function of Yajiao Hadun San. RESULTS: Yajiao Hadun San could reduce the anus temperature of the pyrexia model rat,inhibit the mice's toe and auricular swelling,reduce the times of twisting body sharply and also increase the rate of eliminating colloid carbon in mice. CONCLUSION: Yajiao Hadun San has certain antipyretic,anti-inflammation,analgesic effect and enhance the phagocytosis of mono-karyon macrophage in mice.

The experience of Professor LI Zhidao is introduced in the treatment of mental disorders by "regulating the heart and smoothing the liver". In his experience, the basic prescription includes: penetrating needling technique from Neiguan (PC 6) to Jianshi (PC 5), Ximen (PC 4), penetrating needling technique from Qiuxu (GB 40) to Zhaohai (KI 6) and penetrating needling technique at four groups of acupoints on the gallbladder meridian [Hanyan (GB 4) to Xuanlu (GB 5) and Xuanli (GB 6), Qubin (GB 7) to Shuaigu (GB 8), Shuaigu (GB 8) to Tianchong (GB 9), Tianchong (GB 9) to Fubai (GB 10) and Touqiaoyin (GB 11)]. The manipulation with respiration involved is combined with interactive needling technique so that the patients can feel the weak needling sensation at the local acupoint regions during the treatment. In the paper, the concrete clinical cases are introduced to explain this application and the satisfactory efficacy achieved. The therapeutic method provides a new approach to the clinical treatment of mental disorders.
Acupuncture Therapy ; Adult ; Female ; Heart ; physiopathology ; Humans ; Liver ; physiopathology ; Mental Disorders ; physiopathology ; therapy ; Meridians ; Middle Aged

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.

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.

This study aimed to investigate the anti-apoptotic effects of Tangnaikang (TNK) on islet β cells in Zucker diabetic fatty (ZDF) rats.Six male fa/+ ZDF rats were took as the control group,while other thirty male fa/fa ZDF rats were divided into five groups at random:the model group,the metformin group,the high-,mediumand low-dose TNK groups,depending on their body weight and random blood glucose.Prior to the administration,fasting blood glucose and fasting insulin were measured by drawing blood with inner canthusl.Materials were prepared when administered for six weeks.Fasting blood glucose and fasting insulin were detected again.When the sections of the rat pancreatic tissue were embedded,the morphological changes of the islet were observed via HE staining,and the apoptosis of islet β cell were observed using TUNEL.Positive expression of Caspase-3,the transduction enzyme of cell death signal,was tested by immunohistochemical method.It was found that the fasting blood glucose of the (fa/fa) ZDF rats in the high-,medium-and low-dose TNK group was significantly improved after administration (P ＜ 0.01).The serum insulin of rats in the high-,medium-and low-dose TNK group arised compared with the model group,while the high-and low-dose TNK group showed differences in a statistical sense.Compared with the model group,the HOMA-IR of all the treatment group decreased,while significant difference was presented between the high-dose TNK group and the metformin group.HE staining showed that the morphology of the islet β cell of the rats in all the treatment group was improved.The results of TUNEL showed significantly apoptotic changes on islet β cell of the fa/fa ZDF rats.Compared with the model group,the positive expression of TUNEL in the metformin group and the high-dose TNK group were significantly reduced (P ＜ 0.05).The result of immunohistochemistry method showed that the protein levels of Caspase-3 in the metformin group and the high-dose TNK group decreased (P ＜ 0.05).In conclusion,it was demonstrated that TNK effectively reduced the apoptosis of islet β cells in fa/fa ZDF rats,which presented a protective effect.

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. [