Objective To clone the coding dense granules protein 8(GRA8) from Toxoplasma gondii RH isolate for the potential use in the development of DNA vaccination. Methods Amplifing gene fragment coding GRA8 from the genomic DNA of Toxoplasma gondii RH isolate by means of ploymerase chain reaction (PCR), the gene is inserted into cloning vector pUC-19 digesting with restrictive enzymes and linking react ions. The positive colon is screed on LB plates containing ampicilline and IPTG, Xgal identified by blue-white and restrictive enzyme digestion. The inserted GR A8 gene was recombined with pcDNA3.1(+) eukaryotic expression vector by digestion with restrictive enzyme and linking reactions. The positive coloe is screene d o n LB plates containing ampicillin and identified by restrictive enzymes and link ing reactions. Results The GRA8 gene with about 804 base is specifically amplified from genomic DNA of Toxoplasma GRA8/RH and pcDNA3.1(+)-GRA8 recombinant is successfull y constructed. The sequencing results showed that GRA8 gene of isolate RH and R H from genebank shares quite high homology. Conclusion The gene encoding GRA8 is amplified from genomic DNA of Toxoplasma gene isolate GRA8/RH and pcDNA3.1 (+)-GRA8 recombinant is successfully constructed.

Objective To study the effects ofYiqi Huoxue Jiedu Recipe on immunomodulatory of patients with advanced ovarian cancer.Methods It was performed in a single-arm, open-label study with 40 patients with advanced epithelial ovarian cancer. All patients were treated withYiqi Huoxue Jiedu Recipe lasting for 4 weeks (200 mL, twice a day). The main endpoint was the quantities of CD4+CD25+ cells, CD8+CD28- regulatory T cells and CD8+CD28+ cytotoxic T cells measured by flow cytometry, and the accessory endpoints were variations of symptoms which included pain, fatigue, and numbness etc.Results The quantities of CD4+CD25+ and CD8+CD28- cells significantly decreased compared with those of pre-treatment (P<0.05); the quantity of CD8+CD28+ cell increased significantly (P<0.05). Except for pain, significant improvement showed in fatigue, tinnitus, numbness, poor appetite, abdominal discomfort, insomnia, diarrhea and abnormal urinate (P<0.05).ConclusionYiqi Huoxue JieduRecipe is proved to be effective in regulating immunosuppression advanced ovarian cancer and improving patients’ life quality.

Objective Discbuss the hypertension prevalence rate in the Wuhan male taxi drivers and the dangerous factor to provide the basis for its professional publie health work. Methods The Wuhan male taxi drivers were carried on the healthy physical examination who in the Hubei province medicine medical examination center during 2005-2007 and were carried on the epidemiology statistical analysis. Results The Wuhan male taxi drivers hypertension prevalence rate is 28.28%. That is higher than the common crowd. Hypertension leaves outside the heredity factor and be closely correlated with the age of harnesses, the body weight index, and so on. Conclusions The Wuhan male taxi drivers'hypertension prevalence rate is unoptimistic. We should take the synthesis preventing and controlling measure to reduce this crowd hypertension prevalence rate.

To compare the effects of four different intensive insulin therapies on blood glucose control and vascular endothelial function in newly-diagnosed type 2 diabetes.Patients were randomly divided to accept pre-meal insulin aspart 30 or pre-meal insulin aspart and glargine at bedtime or pre-meal Novolin-R and NPH at bedtime or continuous subcutaneous insulin aspart infusion.Capillary blood glucose determination and continuous glucose monitoring system were carried out,therapeutic time and total insulin dosage were recorded.Ultrasound was used to evaluate the vascular endothelial function.Glucose level,incidence of low glucose,potency ratio of the four groups were similar( P＞0.05 ) ; FMD and NMD were not significantly improved ( P =0.718,P =0.065 ).The short-term efficacy and safety of the four groups are similar.The short-term intensive insulin therapy has no obvious effect on vascular endothelial function.

OBJECTIVE:To investigate the effects of different processed products of Paeonia lactifloria in Shaoyao gancao tang on the content of paeoniflorin in their use.METHODS:The content of paeoniflorin in Shaoyao gancao tang was determined by HPLC in which SHIMA-DCIS ODS C18 was used as chromatographic column with acetonitrile-0.1% phosphoric acid water solution(15∶85) used as a mobile phase at a flow rate of 1.0 mL?min-1.The detective wavelength was set at 230 nm.RESULTS:In Shaoyao gancao tang,the content of paeoniflorin in Radix Paeoniae Rubra was the highest,followed in descending order by Radix Paeoniae Alba Prepara cum Vini,Radix Paeoniae Alba Prepara Flava,Radix Paeoniae Alba sine Cortice and Radix Paeoniae Alba. CONCLUSION:The difference in the content of paeoniflorin appeared when Radix Paeoniae Rubra,Radix Paeoniae Alba Prepara cum Vini and Radix Paeoniae Alba respectively used in combination with Shaoyao gancao tang suggest that Radix Paeoniae Rubra,Radix Paeoniae Alba and its processed products should be used differently based on different syndrome in clinical use of Shaoyao gancao tang.

OBJECTIVE:To explore the spasmolytic effect of baked Radix Vladimiriae on in vitro intestinal muscle in rabbits. METHODS:Magnus in vitro intestinal muscle accumulative dose method was adopted to observe the dose-effect relationship of petroleum ether parts of baked Radix Vladimiriae with in vitro intestinal muscle of rabbits and its impact on the spasm of rabbit intestinal muscle induced by acetylcholine, histamine phosphate and Bacl2. RESULTS:There was a certain dose-effect relationship between petroleum ether parts of baked Radix Vladimiriae and spasmolytic mechanism of rabbit in vitro intestine. It showed the effect of acetylcholine, histamine phosphate and Bacl2. CONCLUSION:The inhibition effect of petroleum ether parts of baked Radix Vladimiriae on normal in vitro intestinal muscle increase as long as dose increase. The inhibition mechanism may be associated with acetylcholine competing with histamine to block M receptor and H1 receptor as well as to inhibit excitability of intestinal muscle.

BACKGROUND: Caspase-3 is well recognized as the key caspase carrying out apoptosis in animal and human brain. To date, a few studies revealed the expression of caspase-3 protein in brains of normal persons and Alzheimer patients but data obtained from rodents exhibited much discrepancy.OBJECTIVE: To investigate the different expression patterns of caspase-3in rodent and monkey brain, and the different expression of caspase-3 in different brain regions and during aging in monkeys.DESIGN: Parallel comparison between means of single variable.SETTING: Institute of Hepatobiliary Surgery, Chinese PLA General Hospital and Department of Anatomy, the Chinese University of Hong Kong.MATERIALS: The experiment was carried out from August, 2003 to February, 2005 in Institute of Hepatobiliary Surgery, Chinese PLA General Hospital and Department of Anatomy, the Chinese University of Hong Kong. Sprague Dawley rats, ICR mice and senescence-accelerated mice (SAM) with ages ranging from postnatal 2, 12, 24 to 48 weeks(n=5 for each age group of different rodents) were included in the present study. All of these animals were supplied by Laboratory Animal Services Center, the hinese University of Hong Kong. Totally 8 rhesus monkeys aged 4 years (n=4) or 20 years (n=4) were selected from the Laboratory Animal Center in Chinese PLA General Hospital [SCXK-(Beijing)2003-002]. Both ro dents and monkeys were female and were raised under standard conditions without any experimental interventions. METHODS: ①Brain tissue samples were taken freshly from both rodents and monkeys and made into homogenate. The expression of caspase-3 pro tein in brains of both rodents and monkeys was investigated with im munoblot. ② The expression levels in monkey brains were exhibited quantitatively with the same method in three brain regions, such as the frontal cortex, hippocampus and cerebellar cortex, for the two age-groups. In vivo distribution patterns of caspase-3-immunoreactive cells were further presented in 3 brain regions of monkeys through immunohistochemistry. MAIN OUTCOME MEATURES: ①Detection of caspase-3 protein with immunoblot in the brain of rodents and monkeys; ② Distribution patterns of caspase-3-immunoreactive cells in 3 brain regions of monkeys. RESULTS: ① Result of detection with immunoblot: The same pattern of caspase-3 protein expression in brain of three kinds of 2-week-old rodents. But the expression was not seen in any other brains of older ages. Caspase 3 was expressed in a relatively high level inboth adult and aged monkey brains, and the amount did not attain to the level in 2-week-old rodents. Caspase-3 Was expressed in the pattern of zymogen (Mr 32 000). The ex pressions of caspase-3 in brains of monkey were not different in ages and brain regions. ②Result of Immunohistochemistry: It was showed that most neurons in the frontal cortex lack detectable caspase-3 immunoreactivity, whereas low to moderate caspase-3 immunostaining be found mainly in pyramidal cells in CA1, CA3 and CA4 subfields of hippocampus. And in the cerebellum, a small number of Purkinje cells were strongly stained in their cytosol and dendrites. Age-related expression pattern of caspase-3 were not found except that in the motor cortex of aged monkeys in which there were a limited number of large pyramidal cells in layer Ⅴ that were strongly stained with caspase-3 antibody.③ Immunoblot procedure revealed that the caspase-3 protein expressed in monkey brains is in the form of zymogen (Mr 32 000) and there is no significant difference in caspase-3 expression level as a function of either brain region or age of animals.CONCLUSION: Unlike rodents in which caspase-3 protein rapidly drops to an undetectable level since animals grow up, the primate expresses caspase-3 constitutively in brain until the late period of lifetime. But there are no significant brain region- or age-related differences in the protein levels in monkey brain.

Objective To investigate the effect of Shugan-Yishen formula on Tamoxifen-resistant MCF-7 cell line(LCC9) through HER2-ERK/MAPK-ERα pathway. Methods Four serums were prepared with the method of serum pharmacology:the serum of Shugan-Yishen formula, Shugan-Yishen formula plus Tamoxifen(TAM), blank and TAM alone. After LCC-9 cells were affected in the above-mentioed serums in vitro, the inhibition rate and the level of mRNA as well as the protein expression of HER2, ERα, ERK/MAPK, p-ERK/MAPK were detected. Results MTT showed that TAM alone group、Shugan-Yishen formula group and the Shugan-Yishen formula plus TAM group had higher inhibition rates at 48th hour, the inhibition rates was(6.61±0.129)%, (47.43±2.24)%, (54.19±3.364)%, compared to the blank group, Shugan-Yishen formula group and the Shugan-Yishen formula plus TAM group had a stronger inhibition on LCC9 at 48th hour (P<0.05), and the combination group was better than Shugan-Yishen formula alone(P<0.05), there was no difference between TAM group compared to the blank group(P>0.05). RT-PCR result showed that compared to the blank group, the expression levels of ESR1 mRNA of group of Shugan Yishen formula plus TAM werehigher by and large at 48th hour (P<0.05), the expression of ESR1 mRNA was(2.282±0.185);and the levels of ERBB2, MAPK3 mRNAwere lower(P<0.05), which levels were(0.71±0.095), (0.336±0.058). Western blotting result showed that Shugan-Yishen Formula plus TAM group could markedly decrease the expression of HER2, p-ERK/MAPK of LCC9 at 48th hour (P<0.05), which the expressions were(0.708± 0.072), (0.228±0.034), and the expression of ER-αelevated(P<0.05). There was no significant difference on ERK/MAPK(P>0.05). Conclusions ①The combination of Shugan-Yishen formula with tamoxifen can inhibit the LCC9 cells. The inhibition effect reaches the peak at 48th hour. ②The mechanism of Shugan-Yishen formula plus TAM combination effects on human breast cancer TAM-resistant cell line LCC9 may improve the sensitivity of ERαby inhibiting the expression of HER2, ERK/MAPK.

Objective To explore the effect of Eight-Section Brocadeon improving extrapulmonary performance and lung function in elderly stable chronic obstructive pulmonary disease (COPD) patients. Methods A total of 120 elderly stable COPD patients were divided into experimental group and control group by random digits table method with 60 cases each. The experimental group received Eight-Section Brocade and the control group took a walk both based on the routine health education. The World Health Organization Quality of Life (WHOQOL)-BREF, 6 min walking test and lung function were conducted before and after intervention 12, 24 and 36 weeks. Results After intervention, the score of each field of WHOQOL-BREF were higher than those before intervention, comparison of two groups of physiological fields Ftime=57.112, Fgroup=10.510, Finteractive=13.653, all P<0.01; psychological field Ftime=240.209, Fgroup=6.201, Finteractive=6.902, all P<0.05;field of social relations Ftime=34.957, Fgroup=7.265, Finteractive=11.554, all P<0.01; environmentfield Ftime=66.431, Fgroup=8.197, Finteractive=18.413, all P<0.01, and there were significant differences in time, groups and interactive. Comparison of two groups of the result of 6 min walking test after intervention, Ftime=327.697, Fgroup=37.181, Finteractive=61.420, all P<0.01. Comparison of two groups of the result of lung function indexforced expiratory volume in one second, Fgroup=5.083, P<0.05, there was significant difference;after intervention the index of Forced expiratory volume in 1 second take up of the expected value, Fgroup=4.141, P<0.05, there was significant difference. Conclusions Eight-Section Brocade can effectively improve the elderly stable COPD patients with exercise endurance and quality of life, improve the extrapulmonary performance, delay the progressive decline of lung function.

Objective: To investigate the level of maternal and infant health literacy and its influencing factors among rural pregnant and lying-in women in Guian New Area, Guiyang City, so as to provide insights into maternal and infant health education. Methods: All pregnant and lying-in women included in health management in township hospitals of Guian New Area from October to December 2021 were selected using a stratified cluster random sampling method. Participants' basic features, maternal and infant health literacy and health education needs were collected using electronic questionnaires, and factors affecting maternal and infant health literacy were identified among pregnant and lying-in women using unconditioned multivariable logistic regression model. Results: A total of 918 valid questionnaires were recovered, and the participants included 325 pregnant women (35.40%), 823 married women (89.65%), 466 women with an educational level of junior high school and below (50.76%), 847 rural household registration (92.27%), 695 women with monthly family income of 3 200 Yuan and less (75.71%), and 267 women with maternal and infant health literacy (29.08%). Multivariable logistic regression analysis identified pluripara (OR=3.418, 95%CI: 1.689-6.919), educational level (diploma, OR=2.901, 95%CI: 1.230-6.842; bachelor and above, OR=6.282, 95%CI: 2.548-15.489), household monthly income per capita (3 201-5 000 Yuan, OR=2.373, 95%CI: 1.499-3.757; 5 001 Yuan and more, OR=7.752, 95%CI: 3.072-19.558), a disease history (OR=3.045, 95%CI: 1.175-5.406), use of maternal and child health manuals (OR=1.881, 95%CI: 1.345-2.632) and emphasizing health education (OR=1.735, 95%CI: 1.122-2.463) as factors promoting for the level of maternal and infant health literacy. In addition, pregnant and lying-in women's major maternal and infant health education needs included infant growth and development knowledge (745 women, 81.15%), management of common infant diseases (719 women, 78.32%), daily infant nursing (651 women, 70.92%) and infant nutrition and feeding (646 women, 70.37%). Conclusions The level of maternal and infant health literacy is low among rural pregnant and lying-in women in Guian New Area, and there is a high demand for health education. High attention should be paid to rural pregnant and lying-in women with a low educational level, low incomes, and primipara.