Objective: To extract and identify the chemical constituents in the roots and stems of Tabernaemontana divaricata. Methods: The solvent extraction and silica column chromatography were used to separate the chemical constituents, and their structures were identified by physicochemical properties and spectra analysis. Results: Six compounds were isolated and their structures were identified as his (2, 3-dihydroxypropyl) octacosanedioate (1), tetratriacontanol (2), palmitic acid (3), glycerol monopalmitatel (4), β-sitosterol (5), and β-daucosterol (6). Conclusion: Compound 1 is a new glyceroesters: compounds 2,3 and 4 are isolated from Tabernaemontana genus for the first time.

BACKGROUND:Now it has cooperation and facilitative effete between myogenic regulatory factors through a long time study. So, gene therapy of double genes of Myod1 and Myog can obtain better effect, and can provide a new way for preventing denervated skeletal muscle atrophy.
OBJECTIVE:To construct eukaryotic co-expression vector carrying Myod1 and Myog genes.
METHODS:Ful-length Myod1 gene and Myog gene cDNA were amplified by reverse transcription PCR, and then inserted into pVAX1 vector after digested to establish the recombined Myod1 and Myog eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and then identified with gene sequencing. The in vitro cultured 3T3 cel s were transfected with pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and the expressions of Myod1 and Myog genes in the 3T3 cel s were detected with western blot assay in order to identify whether the 3T3 cel s could express the target protein correctly.
RESULTS AND CONCLUSION:The sequencing results showed that the sequence length and base sequence of Myod1 and Myog cDNA in eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP were identical with the reported sequences. Myod1 and Myog protein band expressions were detected in 3T3 cel s by western blot after transient transfection. The pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, a eukaryotic co-expression vector of Myod1 gene and Myog gene is successful y constructed.

In the health service field,health resources and health service supple capacity were related with the resource and capacity in economics.Health resources,health service supple capacity and their relationships were important factors for the cooperation and benign interaction of hierarchical medical institutions.However,inversion of health resources and health dislocation of service supply capacity exacerbated the agglomeration degree of patients in the tertiary medical institutions under the impact of residents' freely searching for medical service concept.Vicious interaction and mutual competition among hierarchical medical institutions had become an important obstacle for the implementation of hierarchical medical services.Therefore,its mechanism needed to be clarified while the corresponding policy recommendations should be proposed.

Objective To study the effect of concentration of TCR-specific antigen peptides on priming naive CD4~+ T cells to develop into Th1/Th2 cells or naive CD8~+ T cells to develop into Tc1/Tc2 cells. Methods TCR-specific peptides with series of concentration were co-cultured with routine spleen DCs to activate murine naive CD4~+/CD8~+ T cells. The production of intracellular cytokines IFN-γand IL-4 were measured by flow cytometry. The dividing profile of activated T cells was analyzed by CFSE staining. Results Lower concentration of specific peptides favored Th2/Tc2 polarization while higher concentration benefited Th1/Tc1 polarization. The influence of peptides concentration on Th cells differentiation is higher than that on Tc cells. Conclusion Antigen peptides can stimulate activation of naive T cells in a wide range of concentration. However, with the increase of peptides concentration, activated T cells differentiated grad-ually from Th2/Tc2 to Th1/Tc1, which will provide significant value to control immunized dose of vaccine candidates in animal experiments.

Objective: To investigate the loneliless situation of the elderly, and explore the relationship between loneliness,filial expectations and intergenerational social support among the elderly. Methods: Using UCLA Loneliness Scale, Intergenerational Social Support and Fealty Expectation Questionnaire, we investigated 423 samples of old people aged 60 ～94 of Changsha City. Results: The mean score of loneliness was 39.8 (s =7.3). Ordinal regression analysis indicated that the loneliness of the elderly was significantly related to received adult children social support (OR=2.208, P=0.006),afforded adult children social support(OR=2.056,P=0.01) and filial expectation(OR=1.872,P=0.027),controlled by age, sex, level of education, marriage, level of family income, and residence status. Filial expectation directly affected loneliness and indirectly affected loneliness through received adult children social support and afforded adult children social support. Conclusion: The loneliness of the elderly are significantly related to filial expectation and intergenerational support.

Objective: To extract and identify the chemical constituents in the roots and stems of Tabernaemontana divaricata. Methods: The solvent extraction approach and silica column chromatography were used for separation of the chemical constituents, and their structures were identified by physicochemical properties and spectra analysis. Results: Seven compounds were isolated and their structures were identified as α-amyryn (1), α-amyrin acetate (2), α-amyryl octadecanoate(3), taraxasterol acetate (4), calycosin (5), formononetin (6),and farnisin (7). Conclusion: Compound 1, 3, 4, 5, 6 and 7 are isolated from this plant for the first time, and Compound 3, 4, 5, 6 and 7 are isolated from Tabernaemontan genus for the first time.

This article summarized the characteristics and rules of acupoints grouping recorded in Huang Di Nei Jing (Yellow Emperor’s Canon of Internal Medicine), holding that it had passed important concept of acupoints grouping, formed the embryonic form, and brought up basic grouping types including distant and proximal grouping, and interior-exterior grouping, despite the small number of involved acupoints. Besides, it also paid much attention to other factors that may influence the therapeutic efficacy of acupoints grouping. Therefore, Huang Di Nei Jing has played a crucial role in guiding the development of acupoints grouping theories.

This study was aimed to analyze acupoint selection principle for acute gouty arthritis treatment. Literatures on acupuncture treatment of acute gouty arthritis were collected from CNKI in recent 20 years. Excel software was used in the establishment of database on main acupoints for acute gouty arthritis treatment. On Windows 8 platform, SPSS 17.0 and Clementine 12.0 software were used in the descriptive analysis, association rules analysis and cluster analysis. The results showed that the top three acupionts with the highest frequency of application were Sanyinjiao (SP 6), Zusanli (ST 36), and Y inlingquan (SP 9). The meridian used with the highest frequency was the spleen meridian of foot Taiyin. The result of association rules analysis showed that Sanyinjiao (SP 6)-Xuehai (SP 10)-Quchi (LI 11) was the most relational one. The cluster analysis indicated the top four core cluster groups including Xiangu (ST 43)-Neiting (ST 44)-Xingjian (LR 2), Xuehai (SP 10)-Hegu (LI 4)-Quchi (LI 11), Taibai (SP 3)-Y inbai (SP 1)-Dadu (SP 2)-Fenglong (ST 40), and Zhubin (KI 9)-Zhigou (TE 6)-Gongsun (SP 4)-Bafeng (EX-LE 10). It was con-cluded that the clinical acupoint selection and combination principle in acute gouty arthritis treatment were analyzed through large amount of data. It can provide evidences and references for acupuncture clinical practice.

Background Diabetic retinopathy (DR) seems to be rarely found in high myopia patients with diabetes mellitus.This finding suggests that myopia,especially high myopia,plays a protective effect against DR.It is well-known that the primary pathological base of DR is neovescularization,and the changes of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) are associated with DR.Objective This study was to analyze the expressions of VEGF and PEDF in retina and investigate the influence of high myopia on the pathogenesis of DR.Methods Forty-eight 3-day-old guinea pigs were randomly divided into normal control group,high myopia group,diabetic group and diabetes with high myopia group.Translucent goggles were worn in the right eyes of the guinea pigs for consecutive 8 weeks to induce high myopia,and streptozocin (STZ) was intraperitoneally injected at a dose of 60 mg/kg for four times,every three days for once,to establish diabetic models in the corresponding groups.Twelve weeks after modeling,the animals were sacrificed by excessive anesthesia,and the retinas were isolated for the histopathological examination.Expressions of VEGF and PEDF in the retinas were detected by immunohistochemistry.The use and care of experimental animals conformed to Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The diopter of the normal control group,high myopia group,diabetic group and diabetes with high myopia group were (+0.25±0.07),(-7.50±0.04),(+0.25±0.03) and (-7.50±0.02) D,the fasting glucose of the four groups were (5.3 ±0.1),(5.1 ±0.2),(19.7 ±0.4) and (18.5±0.3)mmol/L,respectively.Compared with the normal control group,the retinas of high myopia group were thin,and the ganglion cells were less.The retinas of diabetic group were loose and oedematous.The retinas of diabetes with high myopia group were thin and oedematous.The expressing level of VEGF in retinas (absorbance value,A) were 128.61 ±5.57,118.24±2.59,155.60±9.70 and 135.15±5.22 in the normal control group,high myopia group,diabetic group and diabetes with high myopia group,respectively,showing a significant difference among the groups (F=17.365,P=0.032),and the VEGF level was significantly lower in the diabetes with high myopia group than that of the diabetic group (t =5.210,P＜0.05).The expressing level of PEDF in retinas (A value) were 145.57± 8.35,149.54±6.20,127.71±2.45 and 137.53±7.38,in the normal control group,high myopia group,diabetic group and diabetes with high myopia group,respectively,with a statistically significant difference among the four groups (F =19.210,P =0.019),and the PEDF level was significantly higher in the diabetes with high myopia group than that of the diabetic group (t=4.521,P＜0.05).Conclusions The expression of VEGF is downregulated and PEDF is upregulated in the retinas of diabetes with high myopia guinea pigs,which may be an accountable mechanism for the low incidence of DR in high myopia eyes.

Objective To analyze the effect of custom abutment and computer aided design and computer aided manufacturing(CAD/CAM) provisional crown on gingival shaping around the implant,and to research the main effect of the custom abutment in the formation of gingival.Methods According to the wishes of patients,fifteen patients were divided into two groups.Gingival molding used CAD/CAM in the observation group(n=7) to design and make custom abutment and provisional crown.The control group(n=8) used the prefabricated abutment and the hand-made self-curing resin temporary crown.Immediately after inserting of final insertion and three months later,pink esthetic score(PES) was calculated in both two groups.Results The labial gingiva curve of the observation group was (1.93±0.27)points,which was higher than (1.56±0.51)points of the control group,there was significant difference between the two groups(t=2.496,P<0.05).After permanent repair of 3 months,the PES scores of the two groups[the observation group (8.71±0.95)points,the control group(8.25±1.04)points]were higher than those of permanent repair day[the observation group (8.29±1.25)points,the control group(7.25±1.04)points],there were statistically significant differences in the total score of PES and permanent repair group permanent repair after 3 months of the day(t=4.290,P<0.05),there was no significant difference between the experimental group(P>0.05).The PES score of the observation group was (8.50±1.09)points,which was higher than (7.75±1.13)points of the control group,but the difference was not statistically significant(P>0.05).Conclusion The effect of the custom abutment on curvature of facial mucosa is significant.The combination of custom abutment and digital provisional crown can achieve better soft tissue stability and ideal aesthetic effect in a short time.