OBJECTIVETo explore the feasibility of inducing human umbilical cord mesenchymal stem cells (HuMSCs) to differentiate into Leydig cells through conditioned medium derived from Leydig cells.

METHODSHuMSCs and Leydig cells were obtained by tissue blocks culture attachment and enzymatic digestion respectively. HuMSCs were induced by conditioned medium of Leydig cells as an experiment group while those before induction were cultured as a control group. The expressions of LHR, 3β-HSD and StAR in the induced HuMSCs were determined by RT-PCR after 3, 7 and 10 days of culture; those of CYP11A1, CYP17A1 and 3β-HSD measured by immunofluorescence staining after 2 weeks; and that of 3β-HSD detected by Western blot after 4 weeks.

RESULTSThe experimental group showed positively expressed LHR, 3β-HSD and StAR at 3, 7 and 10 days, CYP11A1, CYP17A1 and 3β-HSD at 2 weeks, and 3β-HSD at 4 weeks, while the control group revealed negative expressions at all the time points.

CONCLUSIONInduced with conditioned culture medium derived from Leydig cells, HuMSCs are likely to differentiate into steroidogenic cells and eventually into Leydig cells.

Cell Differentiation ; Culture Media, Conditioned ; Humans ; Leydig Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; Umbilical Cord ; cytology

Ambroxol ; therapeutic use ; Dose-Response Relationship, Drug ; Female ; Humans ; Incidence ; Infant, Newborn ; Male ; Respiratory Distress Syndrome, Newborn ; epidemiology ; mortality ; prevention & control

OBJECTIVE To evaluate the difference of efficacy of sublingual immunotherapy on different age patients with allergic rhinitis. METHODS 229 patients with dermatophagoides farina drops allergic rhinitis who have finished SLIT at least 2 years were enrolled and analyzed by retrospective analysis. All patients were divided into different age groups: pre-adolescence(n=107), adolescence(n=53), adulthood(n=69). Indexes for therapeutic effects of SLIT visual analogue scale(VAS) score, rhinitis symptom scores, medication scoreintra- and inter groups were assessed during different stage of treatment(3 months, 1 year and 2 years). Data were analyzed with SPSS 19.0 software. RESULTS Compared with prior treatment, after the SLIT for 3 months, 1 year, 2 years, VAS scores, rhinitis symptom scores, medication scoresof all the patients with allergic rhinitis are significantly decreased; Differences of rhinitis symptom scores(H=0.844, 4.153, 2.669, P>0.05), VAS score(H=1.356, 3.720, 0.313, P >0.05), medication scores(H=1.044, 5.841, 3.399, P>0.05) between groups had no statistical significance at different stage after treatment(3 months, 1 year and 2 years); The differences between 2 years and 1 year of SLIT treatment showed statistical significance in the VAS scores(Z=1.635, P =1.635). CONCLUSION SLIT with dust mites drops had good curative effect and security in patients with allergic rhinitis, which hadno obvious difference between different age groups.

Objective To study the inhibitory effects ofisorhamnetin on six kinds of CYPs of liver in vitro,and the toxic effect on rat hepatocytes Methods This report uses warm incubation of human liver microsomes in vitro to investigate the inhibition of isorhamnetin on 6 kinds of CYPs (CYP2C19,CYP2D6,CYP3A4,CYP2E1,CYP1A2 and CYP2C9),and using HPLC-MS/MS to detect product of metabolism as well as analysing of the pathways of metabolic.At the same time,using rat primary hepatocytes which has low CYPs activity in vitro to explore whether the use of isorhamnetin will cause effects on the ALT,AST and LDH of hepatocytes.Results Isorhamnetin has inhibition effects on CYP2E1 and CYP1A2,the inhibition rate were 59.48％ and 39.91％,respectively.Methylated metabolite is produced after incubating of isorhamnetin and HLMs.The isorhmnetin becomes high polarity and water solubility metabolite 3,3',4',5,7-hydroxyflavone.Isorhamnetin of 30,100 and 300 μmol/L cause a significant rise of ALT and LDH in primary cultured rat hepatocytes cultured (P ＜ 0.01).isorharnnetin of 100 μmol/L cause a rise of AST in primary cultured rat hepatocytes cultured (P ＜ 0.05) and 300 μmol/L cause a significant rise (P ＜ 0.01).It was a dose-dependent manner.Conclusion Isorhamnetin in vitro mainly metabolized by HLMs,and at the same time have a certain inhibitory effect on CYP2E1 and CYP1A2,which may cause the drugs which are metabolized by CYP2E1 and CYP1A2 in vivo accumulation that lead to a series of drug interactions.The results also indicate that heavy use of isorhamnetin cause some adverse effects on hepatocytes,and it was a dose-dependent manner.Individuals need to pay attention to the dose ofisorhamnetin and the potential drug interactions.

ATM: To define the causes of corneal endothelial cell damage, to investigate the preventive methods, and to observe the variety of corneal endothelial cell in glaucoma using confocal microscope.
METHODS: Totally, 143 eyes of 97 patients with different types of glaucoma, and matched normal people were 20 cases, all 40 eyes. The cell density, cell area and cell variable coefficient were measured used confocal microscope. These indicatives of every kind of glaucoma were compared.
RESULTS: The corneal endothelial cell density of normal group was 2 893. 88±255. 026/mm2 , the group of acute angle-closure glaucoma ( AACG ) was 1 674. 11±683.95/mm2 , and the group of open angle glaucoma (OAG) was 2687. 22±391. 87/mm2, the group of chronic angle-closure glaucoma (CACG) was 2706. 97±351. 27/mm2. In all index the average cell density of corneal endothelial and the average area have statistical significance ( F =62.950, 8. 795;P=0. 000), especially the group of AACG.CONCLUSION: The index of corneal endothelial cell in AACG is lower than that of normal. All index in OAG and CACG is difference with that of normal, but the difference has no statistical significance. And the dominant factor of damaged corneal endothelial is the time of intraocular hypertension.

Brain Stem/pathology* ; Death, Sudden/etiology* ; Fatal Outcome ; Humans ; Intracranial Hemorrhages/mortality* ; Leukemia/pathology*

Objective: To investigate the incidence of CMV infection(CMV-I) and CMV related diseases (CMV-D) after allogeneic hematopoietic stem cells transplantation in 70 consecutive allogeneic hematopoietic stem cells transplantation(allo-HSCT) patients and to search for the optimal prophylactic strategy.Methods: Blood samples were monitored using the CMV pp65 antigenemia assay.Of the 70 patients observed,30 patients with chronic myeloid leukemia［CML:CP(27),AP(2),BC(1)］,12 with acute myeloblastic leukemia(AML),10 with acute lymphoblastic leukemia(ALL)and other cases were NHL(3), AA(5), MDS(7), SCLC with pancytopenia (1),CLL(1), and MF (1). Sixty six patients received HLA - identical siblings transplantation and four received tranplants from their HLA- haploidentical donors. Seventy cases included allo-PBPCT (64 cases) , allo-BMT (4 cases) and allo-PB+BMT (2). Before transplantation, all patients and donors received CMV serological examination except 4 pairs of donors/recepients. All 66 patients (3 cases were CMV IgM positive) and 64/66 donors were CMV IgG positive. Results：After transplantation, 64/70 patients developed CMV viremia during monitoring period. Forty three of 70 patients developed CMV-D.Thirty five of them suffered from CMV-associated interstitial pneumonia(CMV-IP). The high peak levels of CMV antigenemia were associated with development of CMV disease　. Close correlation was found between acute graft vs host disease(GVHD) and CMV disease. The patients were followed up for 2 to 24 months. The patients who received preemptive therapy(group A)had significantly better outcome than CMV disease group(group B, P=0.0001). Conclusions: The results suggest that CMV antigenemia has high predictive value for subsequent CMV disease and CMV pp65 antigenemia -guided early therapy has particular advantage for avoiding morbidity and mortality caused by CMV disease.

OBJECTIVE: To study the immunomodulatory effects of the polysaccharide Cistanche Deserticola Y C Ma (CDPS) and its mechanism. METHODS: The immunomodulatory function of CDPS was studied in vitro by observing the proliferation of murine thymus lymphocytes, which was measured with MTT method. The effects of CDPS on cell cycle and thymus intracellular calcium delivering were studied with FACScan flow cytometer. RESULTS: The inhibition function of ISO and DEX and high concentration of TNFgamma on lymphocyte proliferation was decreased with CDPS at higher concentration. It could stimulate the division of thymus lymphocyte and promote thymus intracellular calcium delivering. CONCLUSION: The enhancing effect of CDPS on murine thymus lymphocyte proliferation is related to its promotion on thymus intracellular calcium delivering.

The theory of branch atheromatous disease (BAD) has been commonly underused in clinical practice and research since it was proposed in 1989.In this study,we sought to explore clinical characteristics of its substypes and biomarkers for prognosis of BAD.A total of 176 consecutive patients with BAD were classified into two groups:paramedianpontine artery group (PPA group,n=70) and lenticulostriate artery group (LSA group,n=106).Bivariate analyses were used to explore the relationship between white matter hyperintensities (WMHs),National Institutes of Health Stroke Scale (NIHSS) scores and prognosis evaluated by the modified Rank Scale (mRS) at 6th month after stroke.The differences in prevalence of diabetes mellitus and a history of ischemic heart disease were statistically significant between PPA group and LSA group (x2=8.255,P=0.004;x2=13.402,P＜0.001).The bivariate analyses demonstrated a positive correlation between NIHSS and poor prognosis in patents with BAD and in the two subtype groups,and a positive correlation between WMHs and poor prognosis in the PPA group.It is concluded that a significantly higher prevalence of diabetes mellitus and a history of ischemic heart disease exist in the PPA group than in the LSA group.In addition,high grades of NIHSS scores imply poor prognosis in patients with BAD and in the two subtype groups.Moreover,WMHs are a positive predictor for poor prognosis in patients in the PPA group.