Objective To compare the influence of different coating materials and cultural conditions on the purification and growth of human epidermal melanocytes. Methods The full-thick foreskin, epidermis and cell suspension obtained from human foreskin were cultured in the plates, which were precoated with matrigel or laminin respectively. When having reached 80%-90%confluence, the cells were treated with 0.05%trypsin-EDTA for 4 minutes and resuspended in M254 medium, which were supplemented with G418 and 5-BrdU, respectively. The purity of melanocytes was observed by an immunofluorescence staining with melanocyte markers. Results During the primary culture, the cell suspension generated more cells at faster speed compared with those of skin explants and epidermal specimen. Moreover, the epidermis released cells earlier and proliferated quickly over skin explants. The melanocytes in the plates coated with laminin other than with matrigel displayed faster and better growth. The unwanted keratinocytes and fibroblasts were removed by using differentiation trypsinition combined with supplement of G418 or 5-BrdU. Conclusion Using a plate coated with laminin to culture cell suspension from human foreskin, and via a differentiation trypsinization combined with supplement of small doses of G418 to subculture the cells, is advantageous to the melanocyte purification, without affecting their growth.

Objective To investigate the effects of metformin on serum testosterone (T)levels and ovulation in hyperinsulinemic women Methods Twenty nine patients with hyperinsulinemic anovulation were treated with metformin for 12 weeks Among them 19 had history of clomiphene(CC) treatment with 11 cases failure CC was given again after one month of metformin treatment Serum T, luteinizing hormone,follicle stimulating hormone, insulin(Ins) levels were measured after 4 weeks of treatment Basic body temprature was also observed Results Serum T and Ins levels declined significantly after 4 weeks of metformin treatment [(2 7?1 6) nmol/L Vs (1 9?1 0) nmol/L, (49 2?34 1) mU/L Vs (25 3?23 9) mU/L, P

Objective To investigate the location,fine structure of melanocytes in human fetal scalp hair follicles.Methods The scalp with hair follicles was obtained from a dead fetus of 6 months of age,and divided into two parts.One part was embedded in paraffin,tissue sections were prepared with a width of 7 μm and stained with NKI/beteb,monoclonal antibodies to HMB-45,tyrosinase and tyrosinase-related protein 1(TRP1),respectively.The other part with hair follicles was treated with collagenase type Ⅱ 0.1 g/L and trypsin,then,cell suspension was collected and cultured.After 14-day culture,follicle melanocyte cells (FMC)were separated from keratinocytes by differential trypsinization,and fibroblasts were removed with geneticin.Following three times of pure passage,FMC were seeded and fixed on mica for scanning electron microscopy(SEM)and atomic force microscope(AFM)scanning.Results Histopathological examination showed that NKI/beteb positive cells located at the outer root sheath of human hair follicles,and these cells stained negatively for HMB-45,tyrosinase and TRP1 antibodies.However,in the hair bulb,lots of cells expressed HMB-45,tyrosinase and TRP1 antigens.After fibroblasts and keratinocytes were removed,two kinds of melanocytes remained in the culture:one was small in number and showed abundant melanin,which was lost after subsequent passage;the othgr was large in number and had no melanin initially,but proliferated very rapidly.After three passages,almost all the melanocytes were positive for NKI/beteb.As SEM and AFM showed,most cultured melanocytes appeared fusiform with two(rarely three)dendrites,and the cell body was round or oval with a few melanosomes scattered in but no clear secondary branches on the dendrites.Conclusions The melanocytes in outer root sheath of hair follicles from the fetal scalp are presumed as melanocyte stem cells or their progenies.In vitro,these cells proliferate very rapidly during early phases,but the morphology and function of them still remain immature,which is unfavorable for melanosome transport.

OBJECTIVESTo study the influence of insulin on IGF-I and IGFBP-I secretion of the human endometrial stromal cells.

METHODSLate proliferative phase endometrial stromal cells were isolated from endometrium tissues and then cultured for 24 h in Hams F-12 only as a control and in Hams F-12 with different concentrations of estradiol (E2) and insulin (INS) as treated groups. Simultaneously, the endometrial stromal cells from late secretory phase endometrium were cultured for 24 h in Hams F-12 only as a control and in Hams F-12 supplemented with different concentrations of progesterone (P) and insulin as treated groups. After 24 h of culturing, the mediums were collected for either IGF-I or IGFBP-I assays.

RESULTThe concentrations of IGF-I in medium from cultured endometrial stromal cells in the proliferative phase were 0.78 +/- 0.47 ng/ml in the hormone-free control group; 1.44 +/- 0.59 ng/ml and 1.39 +/- 0.33 ng/ml in 100 pg/ml E2 group and 20 microU/ml INS group, which was higher than that of the control group (P < 0.05 and P < 0.01, respectively). The IGF-I concentration in the 100 microU/ml INS group was 2.03 +/- 0.53 ng/ml, which was higher than that of the 20 micro U/ml INS group (P < 0.01). Levels of IGF-I in the 100 pg/ml E2 plus 20 microU/ml INS group was 2.18 +/- 0.36 ng/ml, which was significantly higher than that of the 20 microU/ml INS and 100 pg/ml E2 group (P < 0.01), but lower than that of the 100 pg/ml E2 plus 100 microU/ml INS group (3.42 +/- 0.75 ng/ml), P < 0.01. The concentration of IGFBP-I in medium from cultured endometrial stromal cells in the secretory phase was 2.50 +/- 1.39 ng/ml in the hormone-free control group and 5.44 +/- 2.09 ng/ml in the 10 pg/ml P group, which was significantly higher than that of the control (P < 0.01). IGFBP-I concentration in 20 microU/ml INS group was 0.16 +/- 0.58 ng/ml, which was lower compared with control, but higher compared with the 100 microU/ml INS group (P < 0.01). The level of IGFBP-I in the 10 ng/ml P plus 20 microU/ml INS group was 2.10 +/- 1.17 ng/ml, lower compared with the 10 ng/ml P group, but higher compared with the 10 pg/ml P plus 100 microU/ml INS group, P < 0.01.

CONCLUSIONSInsulin can stimulate basal (without hormone) and E2-stimulated IGF-I secretion in cultured stromal cells from human late proliferative endometrium in a dose-dependent manner. Insulin can suppress basal (without hormone) and P-stimulated IGFBP-I secretions in cultured stromal cells from human secretory endometrium in a dose-dependent manner.

Cells, Cultured ; Dose-Response Relationship, Drug ; Endometrium ; cytology ; drug effects ; secretion ; Estradiol ; pharmacology ; Female ; Humans ; Insulin ; pharmacology ; Insulin-Like Growth Factor Binding Protein 1 ; secretion ; Insulin-Like Growth Factor I ; secretion ; Progesterone ; pharmacology ; Stromal Cells ; drug effects ; secretion

Objective To investigate the pathological types and features of lymph nodes in human immunodeficiency virus(HIV)/acquired immune deficiency syndrome(AIDS)patients with superficial lymphadenectasis.Methods The tissues of lymph nodes were obtained from 151 HIV/AIDS patients with superficial lymphadenectasis for pathological examination.The pathological results were observed by light microscope after Hematoxylin-Eosin(HE),acid-fast,periodic acid-Schiff (PAS),and digested-PAS(D-PAS)staining.The pathological results of lymph nodes were described and the correlation between pathological changes and CD4+T lymphocyte count was analyzed.Chisquare test was used for the statistic analysis.Results The benign lesions were found in 145 patients (96.0%),while the malignant tumors were found in 6 patients(4.0%).The pathological findings in the 151 HIV/AIDS patients included tuberculosis(72 patients),lymph node reactive hyperplasia(34patients),lymphatic fungal infections(23 patients,including penicillium diseases in 19 cases),AIDSrelated lymphadenectasis(14 cases),non-Hodgkin lymphoma(5 cases),benign fibrous histiocytoma (1 case).In addition,there were 83 patients(55.0%)with CD4+T lymphocyte count lower than 100×106/L.The frequency of penicillium diseases was higher in patients with lower CD4+T lymphocyte count(x2=7.757,P=0.021).Conclusions The major reasons for superficial lymphadenectasis in HIV/AIDS patients are infectious diseases,such as tuberculosis and fungal infections,which are common in patients with lower CD4+T lymphocyte counts.Non-Hodgkin lymphoma is the most common malignant tumor in this patient population.

Objective To investigate the characteristics of diet in patients with gestational diabetes mellitus(GDM) and analyze their dietary nutrition.Methods Clinical data of 978 cases of pregnant women in our hospital were collected and divided into GDM group(458 cases) and the healthy control group (520 cases).Dietary questionnaire was designed and diet characteristics were analyzed retrospectively.Daily calories and nutrients intake were calculated by software calculation.Results There were significant differences on the dietary pattern between the two groups:daily coarse grains:(35.12 ±4.87) g vs (46.35 ±5.23) g;seafood:(22.10 ± 8.45) g vs (39.34 ± 12.45) g;grilled:(6.72 ± 1.76) g vs (3.45 ±0.34) g;fried:(25.34 ± 7.84)g vs (10.56 ±4.76) g;sausage:(5.38 ± 1.81) g vs (3.45 ± 0.45) g;dessert type products intake:(50.41 ±20.76) g vs (35.86 ± 10.45) g (t =2.582,10.346,15.452,20.239,18.195,8.472,respectively,P ＜0.05).There was only significant difference between the two groups on calcium supplements in daily intake of nutritional supplements(GDM group 43.89％,control group 72.69％,x2 =83.660,P ＜ 0.001),and there was no significant difference on the rests(all P ＞ 0.05).Vitamin A,B1,B2,C and calcium,iron,zinc,selenium daily intake in GDM patients were less than those in the control group,but there was significant difference only in calcium compared with the control group (GDM group (785.2 ± 194.5) mg,control group (945.5 ± 245.2)mg,t =2.795,P =0.005).Multivariate logistic regression found that excessive intake of fried class (OR =3.357,95％ CI =2.012-11.340),barbecue (OR =2.652,95％ CI =1.834-8.578),sausage type products (OR =2.518,95％ CI =1.743-7.421) etc.were dietary factors of GDM (all P ＜ 0.05).Incidence of GDM was significantly increased if taking more.While the increased intake of coarse grains (OR =0.632 95％ CI =0.134-1.489),seafood class(OR =0.408,95％ CI =0.105-1.189) can reduce the occurrence of GDM(all P ＜ 0.05).Conclusion Many unscientific ways of eating involved in pregnant women with GDM.Dessert,sausage,grilled,fried products intake increased the risk factors for GDM.Coarse grains,seafood intake were the protective factors of GDM.Timely adjustment of maternal diet and adequate calcium supplementation played an important role in the prevention of GDM.

Objective To investigate the problem of adverse effects in common AIDS patients and AIDS patients with tuberculosis after highly active antiretroviral therapy (HAART). Methods The case group was composed of 106 patients with both AIDS and tuberculosis. The control group was composed of 134 common AIDS patients. The rates of adverse effects and the increase of CD4 + T cell count in those groups after first year HAART were observed and compared. Results The rates of adverse effects in the case group was 36. 8% ,which was more than that in the control group (26. 9%), but the difference was not significantly different(x2 =2.715, P =0. 099). The count of CD4+ T cell in most of the patients was increased after HAART (P ＜ 0. 01). The increase of CD4 + T cell count in the case group [(147.2 ±137.6)/μl] was higher that in the control group[(142. 1 ± 127. 0)/μl after six months HAART vs. (166. 5±133. 1)/μl in case group], and it was lower than that in control group after nine months HAART [(172.7±107.5)/μl], however the difference was not significant(P ＞0.05). Conclusions HAARTcould reconstruct the immunition of AIDS patients. The increase of CD4 + T cell count did not show significant difference between common AIDS patients and AIDS patients with tuberculosis after HAART. AIDS patients with tuberculosis might not increase the risk of development of adverse effects during HAART.

Objective:To search and summarize the best evidence for blood pressure management in patients with intracerebral hemorrhage to provide evidence for blood pressure management in these patients.Methods:PIPOST model was used to establish evidence-based questions; with "intracerebral hemorrhage/cerebral hemorrhage/ICH/HICH/stroke" AND "blood pressure/hypertension" as English search terms, and with "脑出血/颅内出血/脑卒中"+"血压" as Chinese search terms, the official websites of related associations and domestic and foreign databases, such as GUIDE.com, UpToDate, Cochrane Library, CNKI, VIP Chinese journal Service platform, and Wanfang Data Knowledge Service platform, were searched. Two researchers independently evaluated the quality of the literature, including guidelines, systematic reviews, and expert consensus, and extracted the evidence.Results:A total of 17 articles were selected; 2 were clinical decision type, 7 were guideline type, 7 were systematic review type, and 1 was expert consensus type. Twenty-six pieces of evidence were summarized into 5 aspects: timing and methods of lowering blood pressure, benefits and risks of lowering blood pressure, drug selection and selection methods, recurrence prevention of stroke, and matters for attention.Conclusion:It is necessary to pay attention to blood pressure management in patients with intracerebral hemorrhage; in the process of evidence application, it's suggested to make an appropriate blood pressure management plan for the patients according to clinical situation.

OBJECTIVETo explore the safety and efficacy of Shaobei injection in the management of I(-II( degree rectal prolapse.

METHODSA total of 80 patients eligible for the inclusion criteria were divided into 2 groups: 40 patients in the treatment group (treated with Shaobei injection) and 40 cases in the control group (treated with Xiaozhiling) respectively. The short-term efficacy was identified by the length of rectal prolapse and the depth of rectocele demonstrated by the defecography while the long-term efficacy was evaluated by the length of rectal prolapse. In addition, the safety was assessed by the occurrence of postoperative complications.

RESULTSThe variation of the lengths of rectal prolapse and the depth of rectocele demonstrated by the defecography at the sixth month follow up in the two groups did not reach significant difference (P>0.05). After 2 year follow up, 37 patients (92.5%) in the treatment group and 35 cases(87.5%) in the control group was cured. Therefore, there was no significant difference in the long term efficacy between the two groups (P>0.05). The adverse events in the therapy group(10%) was less than that in the control group (45%)(P<0.01).

CONCLUSIONSShaobei injection in the management of I(-II( degree rectal prolapse has a similar efficacy of short-term and long-term higher safety compared to Xiaozhiling injection.

Defecography ; Female ; Humans ; Injections ; Postoperative Complications ; Rectal Prolapse ; drug therapy ; Rectocele

Objective:To develop a professional grief scale for medical postgraduate students(PGS-MPS) and test its reliability and validity.Methods:Based on the integrated model of professional grief, the initial item pool was formed after literature analysis, semi-structured interviews, group discussion, and 5 experts were invited to assess the relevance of the items and provided revision suggestion.A total of 506 valid questionnaires were collected.SPSS 25.0 and AMOS 26.0 were used to test its reliability and validity.Results:The PGS-MPS included 41 items, including the professional grief reaction subscale and disenfranchised grief subscale.The professional grief reaction subscale included emotional reaction (14 items), physiological and cognitive reaction (20 items), and the cumulative variance contribution rate of the two factors model was 52.908%.Disenfranchised grief subscale included 7 items, with a single dimension and a variance contribution rate of 51.730%.Confirmatory factor analysis indicated the model fitting well.The Cronbach’s α coefficients for the professional grief reaction subscale and disenfranchised grief subscale were 0.947 and 0.850, respectively.Conclusion:The PGS-MPS has good reliability and validity and can be used to evaluate professional grief of medical postgraduate students.