The afferent connections to the periaqueductal gray in the rat were examined by means of retrograde axonal transport method using horseradish peroxidase (HRP).The microinjection syringe needle or micro-electrophoretic pipette was inserted stereotaxically by an oblique approach through the cerebellum. HRP was introduced into the dorsal, lateral, ventrolateral areas of periaqueductal gray or periventricular gray of ventriculus Ⅲ at posterior commissure level respectively.The results were as follows:1. Injections of HRP in the dorsal area of periaqueductal gray(PAG).Numerous labeled cells were consistently found in the paraventricular nucleus, dorsomedial and ventromedial hypothalamic nuclei, posterior hypothalamic nucleus, medial mamillary nucleus, zona incerta, lateral and ventral nuclei of periaqueductal gray, superior colliculus, inferior colliculus, dorsal raphe nucleus and periventricular gray of ventriculus Ⅳ. In some cases, a smaller number of cells was found in the zona reticulata of the substantia nigra, and lateral hypothalamic nucleus. In one case, the enzyme was injected into the dorsal area of PAG, which diffused to the superior and inferior colliculi, resulted in labeling of Forel's field and reticular nucleus of thalamus.2. Injection of HRP in the lateral area of PAG.HRP-positive cells were observed within the bed nucleus of the stria terminalis, paraventricular nucleus, anterior hypothalamic nucleus, ventro-medial hypothalamic nucleus, medial thalamic nucleus, nucleus interstitialis magnocellularis, commissural posteriors, zona incerta, ventral nucleus of PAG, zona reticulata of substantia nigra, formatio reticularis of mesencephalon, dorsal raphe nucleus, and periventricular gray of ventriculus Ⅲ and Ⅳ. In some cases, a small number of cells was found in the supraoptic nucleus, superior and inferior colliculi.3. Injection of HRP in the ventrolateral area of PAG.In all cases the greatest number of HRP-positive neurons was shown in the supraoptic nucleus, paraventricular nucleus, periventricular gray of ventriculus Ⅲ, lateral nucleus of PAG, zona reticulata of substantia nigra, dorsal raphe nucleus and locus coeruleus. The labeled cells were mainly located ipsilaterally. But the labeling of supraoptic nucleus was observed bilaterally. In one case the enzyme diffused to the reticular formation, a smaller number of cells was found in nucleus cuneifomis and mesencephalic reticular formation of more caudal position.4. Injection of HRP in the periventricular gray of ventriculus Ⅲ.Following the injections of HRP in the periventricular gray of ventriculus Ⅲ, numerous HRP-positive cells were consistently observed in the bed nucleus of the stria terminalis, supraoptic nucleus, paraventricular nucleus, ventromedial nucleus of hypothalamus, medial nucleus pars medialis of thalamus, zona incerta, ventral nucleus of PAG, zona reticulata of substantia nigra, dorsal raphe nucleus and locus coeruleus.

Objective To investigate the expression and significance of hypoxia-inducible factor 1 alpha (HIF- 1α), vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) in tissues of condy- loma acuminatum (CA) in pregnant women. Methods Tissue specimens were obtained from the lesions of 30 pregnant women with CA and 30 non-pregnant women with CA, and from the vulva of 15 normal preg- nant women. By immunohistochemical staining, the expressions of HIF -1α, VEGF and Ang-2 were detected in these specimens. Results The expression rates of HIF -1α,VEGF and Ang-2 were 86.67%, 93.33% and 83.33% in pregnant women with CA, respectively, 63.33%, 66.67% and 53.33% in non-pregnant women with CA, respectively, and 0, 6.67% and 0 in normal women, respectively. Enhanced expressions of HIF -1α,VEGF and Ang-2 were observed in pregnant women with CA compared with the latter two groups (P < 0.05). In pregnant women with CA, a significant correlation was noted between the expression of HIF -1α and VEGF (r = 0.412, P < 0.01) and between the expression of Ang-2 and VEGF (r = 0.460, P < 0.01). Further more, the expression of HIF -1α, VEGF and Ang-2 positively correlated with each other in non-preg- nant women with CA. Conclusion In tissues of CA in pregnant women, HIF-1α,Ang-2 and VEGF are over expressed, which may be related to angiogenesis.

Objective To investigate the formation and elimination of photoproduct in epidermal cells from BALB/c mice irradiated with ultroviolet B, and to observe the interference by baicalin in it. Methods BALB/c mice were randomized into 6 groups, I.e., blank control group receiving no exposure or protection, baicalin group receiving protection with baicalin, acetone group receiving acetone pretreatment, UVB group receiving UVB irradiation but no protection, UVB + baicalin group receiving UVB irradiation and protection with baicalin, UVB + acetone group receiving acetone pretreatment and UVB irradiation. Baicalin was applied at 1 mg/cm2 on the back of mice for 3 days in baicalin group and UVB + baicalin group. Twenty hours after the last application, UVB irradiation of 180 mJ/cm2 was given to mice in UVB group and UVB + baicalin group. Skin specimens were obtained from the tested sites at 1, 24, and 48 hours, respectively, after the irradiation. Cyclobutane pyrimidine dimers (CPD) was detected in the specimens with immunohistochemical staining and Southwestern dot blotting. Results CPD was observed only in irradiated mice. The relative content of CPD in epidermal cells 1, 24 and 48 hours after the irradiation was (100±5.22)%, (75.34±8.22)% and (42.11±3.24)%, respectively, in UVB group, (81.45±5.22)%, (32.14±6.33)% and ( 5.21±3.15 )% respectively, in UVB+baicalin group, ( 106±8.21 )%, (70.23±4.13 )% and (41.22±4.21)%, respectively, in UVB + acetone group. A significant difference was observed in the relative content of CPD between UVB group and UVB + baicalin group at 1, 24 and 48 hours after the irradiation (P<0.05, 0.01, 0.01, respectively). Conclusions Taken together, these results suggest that topical baicalin application mitigates DNA photo-damage. Baicalin is therefore a promising protective substance against UVB radiation.

Objective:To prepare morphine sulfate sustained-release suppositories and determine the content by HPLC. Methods:An Eclipse XDB-C18 (150 mm × 4. 6 mm,5 μm) chromatographic column was used, methanol-heptane sulfonic acid sodium acetate solu-tion(2. 02 g sodium heptanesulfonate was dissolved in appropriate amount of water and 5ml glacial acetic acid was added, and then water was added to 1000 ml, shaken up) (50 ∶50) was used as the mobile phase, the flow rate was 1. 0 ml·min-1 , the detection wavelength was 233 nm, the column temperature was 25℃ and the sample size was 10μl. Results:The average content of morphine in 3 batches of samples was 99. 9%, the linear range of 4. 18-86. 60μg·ml-1 was good (r=0. 999 3), and the average recovery was 100. 6% (RSD=1. 58%, n=9). Conclusion:The method is sensitive, rapid and accurate, and suitable for the quality control of morphine sulfate sus-tained-release suppositories.

Aim Toinvestigatethespecificbinding sites that HDAC1 can be recruited to regulate the tran-scriptional activity of p21 WAF1/CIP1 promoter in the breast cancer MCF-7 cells.Methods ThebreastcancerMCF-7 cells in logarithmic growth phase were starved with FBS free medium for 24 hours,and treated with 20 μmol·L-1 SAHA(S group)or 0. 625 nmol·L-1 Leptin(L group)for 24 hours,and the cells that were cultured in the complete RPMI 1640 medium without any treatment were assigned as control group (B group).The DNA-ChIP was followed the manufactur-er′s protocol for the assay.The cell lysis was prepared and incubated with anti-HDAC1 antibody overnight at 4℃.DNA fragments binding anti-HDAC1 antibody were gathered and purified.The relative expression level of DNA fragments from TSS to the upstream of the p21 WAF1/CIP1 promoter region(+2 ~-4000 bp)bind-ing with antibody was detected by Real-time PCR and analyzedby2-ΔΔCTmethod.Results InBgroup, HDAC1 had high affinity with the f1 and f 8 fragmentsof p21 WAF1/CIP1 promoter compared to the other fragemts,and showed the highest affinity with the f8 fragment.In S group,the binding ability of HDAC1 to the f1 ~f10 fragment of p21 WAF1/CIP1 promoter was sig-nificantly lower than that of the control.The binding activity of HDAC1 to f8 fragment was the lowest,while reversing to reach the peak after leptin treatment.Con-clusions HDAC1canberecruitedtop21WAF1/CIP1pro-moter by the cell proliferation signal during the prolifer-ation of breast cancer MCF-7 cells.The DNA fragment from -2800 to -3200 bp in the upstream of p21 WAF1/CIP1 promoter is the target functional region for the binding with HDAC1 .

With the development of assisted reproductive technology , it brings the gospel to many infertility families , and it makes a new challenge to the moral and legal at the same time .Ministry of Health issued the notice on carrying out the special action to manage assisted reproductive technology .By thinking about the ethical and le-gal issues, this paper tried to explore a new response mode to advance the development of assisted reproductive technology and to ensure social harmony and stability .

In order to understand the prevalence of the obesity in children and adolescence and provide the basic information for developing scientific strategy for obesity prevention,this paper described the common methods to evaluate the obesity of children and adolescence,and its prevalence,distributions of population and tendency of changing.The paper also analyzed the effects of sex,age,region and socioeconomic status on obesity.

Objective To explore the clinical characteristics and risk factors of healthcare-associated infection (HAI)in patients with severe viral hepatitis,so as to provide basis for clinical prevention and treatment of HAI. Methods Clinical data of 193 patients with severe viral hepatitis and admitted to a hospital from January 2012 to November 2015 were analyzed retrospectively,clinical characteristics and risk factors for HAI in patients were evaluated,all data were statistically analyzed.Results Among 193 patients,75 developed 112 times of HAI, incidence and case incidence of HAI were 38.86% and 58.03% respectively.The main infection sites were abdomi-nal cavity,lower respiratory tract,and gastrointestinal tract,infection rates were 33.93%,25.00% and 16.96%respectively.A total of 58 strains of pathogens were isolated,including fungi,gram-negative bacteria,and gram-positive bacteria,accounting for 53.45%,36.21%,and 10.34% respectively.Mulvariate logistic regression analy-sis showed that underlying diseases,use of glucocorticoid,invasive procedure,and white blood cell count<4.0×109/L were independent risk factors for HAI in patients with severe viral hepatitis (P<0.05).Conclusion Incidence of HAI in patients with severe viral hepatitis is high,risk factors are multiple.It is necessary to take targeted prevention measures during the treatment,so as to reduce the incidence of HAI.

Objective To research the calibration methods of the electrosurgical analyzer which is applied in calibration for production and use of electrosurgical generator. Methods Two methods were put forward,the first one is indirect measurement,the second one is comparing with congeneric device,and these two methods in theory and practice to process real data were analyzed. Results The device with the methods are tested and analyzed,and then,the test error or uncertainty of electrosurgical analyzer is used in analyzing and processing. Conclusion The test error or uncertainty of leakage current and peak factor can also get by these two methods,and these methods could be used as calibration method in production and usage of electrosurgical generator or method for tracing the source.

Objective To obsOrvO thO application OffOct of modifiOd midwifOry tOchniquO in natural childbirth of low - risk primipara, thus to providO tOchnical support for thO dOlivOry of continuous quality improvOmOnt. Methods From JunO 2015 to SOptOmbOr 2017, 120 low-risk primipara with natural childbirth in thO MatOrnal and Child HOalth CarO Hospital of Zhoushan wOrO sOlOctOd and randomly dividOd into two groups by computOr random coding mOthod, with 60 casOs in Oach group. ThO control group rOcOivOd traditional midwifOry tOchniquO, and thO obsOrvation group adoptOd improvOd midwifOry tOchniquO. ThO pOrinOum incision ratO, pOrinOal lacOration classifica-tion, thO condition of matOrnal and infant, and pOrinOum incision and not cut matOrnal blOOding amount, thO sOcond availablO labor of thO two groups wOrO comparOd. Results ThO incision ratO was lowOr in thO obsOrvation group (53.33% ),and thO ratO of non-split wound was highOr(76.67% ),and thO incidOncOs of advOrsO OvOnts in thO obsOrvation group wOrO lowOr than that in thO control group(all P<0.05). ThO total amount of blOOding in thO obsOr-vation group was lOss [(151.05 ± 64.59)mL], and thO sOcond labor timO was shortOr[(24.63 ± 19.58)min] than thosO in thO control group (t=2.21,4.32,all P<0.05).Conclusion Application of improvOd midwifOry in natural dOlivOry of low-risk primipara is OffOctivO.