In order to validate and estimate the capability of BMP9 to induce osteogenic differentiation of multipotent stem cells, three multipotent stem cells(C3H10, MEFs and BMSC) were used as target cells, and BMP9 was introduced into these cells by using recombinant adenoviruses assay, the effect of BMP9 on osteogenic differentiation of multipotent stem cells was demonstrated by using luciferase reporter assay, alkaline phosphatase(ALP) quantitative assay, calcium deposition assay, real time PCR, animal experiment and histological staining assay.The results demonstrated that BMP9 can induce ALP expression of C3H10, MEFs and BMSC by a dose dependent manner.BMP9 can also stimulate calcium deposition of C3H10 and MEFs in vitro, the osteogenic markers(ALP, Runx2, osteopontin, osteocalcin) were increased after stimulated by BMP9.BMP9 can activate canonical TGF?-Smad pathway, and promote the expression of osteogenic master gene Runx2.The animal experiment and histological staining assay show that BMP9 can induce ectopic bone formation in naked mice.To sum up, BMP9 is a more powerful cell factor to induce osteogenic differentiation of multipotent stem cells.

Bronchi ; cytology ; Cell Culture Techniques ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology

ABSTRACT:Objective To construct V-set and immunoglobulin domain containing 4 (Vsig4)nanobodies (Nbs) as specific macrophage probes so as to use them as molecular probes of macrophagocytes.Methods A nanobody phage library was generated by using peripheral blood lymphocytes isolated from an alpaca immunized with recombinant Vsig4 protein.After three rounds of selection against recombinant Vsig4.The Nbs were subjected to sequencing and genome alignment to obtain VHH sequence.Nbs were isolated and tested for Vsig4 specificity in an ELISA using recombinant Vsig4.The affinity capacity of Nbs was verified by the cell line stably expressing Vsig4. Results A nanobody phage library with an estimated 7.27 × 107 clones with 70% insertion was successfully constructed.Totally 1 3 6 Vsig4-positive clones were sequenced and aligned according to different CDR3 sequences. In summary,1 5 Vsig4 nanobodies were obtained and grouped into 3 different CDR3 epitopes.The affinity of representing nanobody and Vsig4 was analyzed via ELISA;Nb1 1 9 showed the highest affinity against both recombinant and native Vsig4.Conclusion We successfully constructed and screened Vsig4 specific nanobody number 1 1 9 with high affinity and specificity.It can help with macrophage detection and in vivo monitoring.

Objective To evaluate the pharmacological effect of Zhixie Tuire Tablets(ZTT) on human rotavirus RV709 in host cells in vitro.Methods By improved methods of methyl thiazolyl tetrazolium colorimetry assay(MTT) and cytopathic effect(CPE),the anti-rotavirus effect of ZTT on RV709 was evaluated.Results The half toxic concentration(TC50) of ZTT for MA104 was 0.49699 mg/mL.ZTT neither can directly kill human rotavirus,nor prevent human rotavirus from invasion of host cells.The halft inhibitory concentration of ZTT was 0.00519 mg/mL for RV709.Conclusion In-vitro anti-rotavirus mechanism of ZTT is related to the inhibition of the biological synthesis of rotavirus in host cells.

Objective To probe into the therapeutic mechanism of Zhixie Tuire Tablet (ZTT) for rotavirus-induced diarrhea. Methods We established the 4-day old Kunming neonatal mice model by infecting group A G3 type of rotavirus,and then gave the inoculated mice with ZTT at the dose of 0.78 g/kg for three days. Fluorescent quantification PCR method was used to detect the changes of sodium glucose transporter SGLT1 mRNA level and radio-immunity method was used for the examination of PGE2 level in neonatal mice small intestine. Results SGLT1mRNA level was lower in ZTT group than that in the normal group,the difference being significant (P 0.05). PGE2 level in ZTT group was higher than that in the normal group,but lower than that in the model group (P

Objective Few prospective data are currently available on acute gastrointestinal hemorrhage (AGIH) as a complication in acute kidney injury (AKI).The aim of the present study was to find out clinical characteristics,incidence,etiology,risk factors,and outcome of AGIH in patients with AKI.Methods We performed a prospective study on an inceptione cohprt of 512 patients admitted for AKI in our hospital.Data on clinical risk factors for bleeding,frequency of occurrence of AGIH,in-hospital mortality were collected,and independent predictors of AGIH were identified.Results A total of 53 patients had AGIH as a complication of AKI,and 45 were upper AGIH.Fifteen patients had clinically severe bleeding.Independent baseline predictors of AGIH were severity of illness,cardiac failure,mechanical ventilation,low platelet count,chronic hepatic disease,liever cirrhosis,severe AKI.Inhospital mortality was 52.8％ in patients with AGIH,and 22.2％ in the other patients.AGIH was significantly associated with an increase in hospital mortality.Conclusions AGIH are frequent complications of AKI.In this clinical condition,AGIH is more often due to upper gastrointestinal bleeding and is associated with a significantly increased risk of death.Both renal and extrarenal risk factors are related to the occurrence of AGIH.

Objective:To investigate the characteristic and the hearing change of hearing thresholds of the infants failing in the hearing screening.Method:802 infants(1179 ears) with hearing screening record were reviewed in the study. They had hearing screened by otoacoustic emissions (OAE).However, they failed in the first or & and second hearing screening. Auditory brainstem response (ABR) , distortion product otoacoustic emissions(DPOAE) and acoustic immittance measurement were examined. Subjects with abnormal hearing-evaluation were retested during the following 1-3 months (before 6-month-old). To analysis the characteristic and the hearing change of hearing thresholds of the tests.Result:Among 802 cases(1179 ears),the ratio of single-ear(53.2%) is higher than both-ears(39.5%), P<0.05. And there are no significance between the two groups in the ration of severe abnormal hearing-evaluation ;25.8% infants once suffered from hypercholesterolenia failed in the hearing screening,which was higer than the other known risk factors.P＜0.05; Threatened abortion group has found no cases of severe hearing abnormalities; The ratio of severe abnormal hearing-evaluation in family history hearing loss group (40.9%) was higher than other high-risk factors, followed by two or more risk fators group(10.8%);56.9% of the infance who did not pass the hearing screening were found none risk factors, 42.3% of the infances who had high risk factors was normal hearing level;96 cases(138 ears) undergone twice assesement, in morderat disorder group, more cases(74.3%) changed better , and less disorder group(40.9%), severe disorder group (33.3%).Seven ears became worse; Among abnormal hearing induced by different high-risk factors, the proportion of hearing improvement in children with moderate abnormal hearing was higher than that in children with slight abnormal hearing. Ten cases(12 ears) diagnosised auditory neuropathy.Conclusion:The important factors make the follow-up decision incluing perinatal history,hearing level and age.Maybe some high-factors we did not realized. Gene screening should be paied more attention in the future work.

Objective To assess the short-time economics of various glutamine dipeptide-enriched parenteral nutrition (PN) for patients undergoing elective surgery for gastrointestinal tumors, with an attempt to provide evidence for decision makers on clinical nutrition support.Methods A prospective cohort study was designed.From payer/disburser''s perspective, a cost-effectiveness decision-tree model was developed to assess the clinical outcomes and short-time economic effects of glutamine dipeptide-enriched PN that used in different time points (early postoperative or perioperative).Cost-effectiveness analysis, cost-utility analysis, and incremental cost-effectiveness analysis were adopted in the decision-tree model.One-way sensitivity analysis was performed to determine the robustness of the results.Results Totally 107 patients were included.There was no significant difference between the perioperative alanine(Ala) glutamine(Gln) nutrition support (group A) and early postoperative Ala-Gln nutrition support (group B) in the ratio of 5％ weight declines on the 8th day after surgery and infection-related postoperative complications (72.1％ vs.78.1％, χ2=0.509, P=0.498 and 2.32％ vs.4.69％, χ2=0.060, P=0.806).The levels of prealbumin (PA) and albumin(Alb) and the level of total lymphocyte count(TLC) also the time of recovering gastrointestinal function, length of stay nutritional discharge index(LOSNDI), and direct costs were significantly different [PA:(208.19±56.92)mg/L vs.(187.97±62.05)mg/L, t=2.283,P=0.039;Alb:(33.82±3.91)×109 vs.(31.96±4.57)×109, t=2.184, P=0.036;TLC:(1.19±0.55)×109 vs.(0.89±0.66)×109, t=2.461, P=0.015;the time of recovering gastrointestinal function(3.06±0.28)d vs.(3.39 ± 0.34)d, t=-3.675, P=0.000;LOSNDI:(16.84±2.92)d vs.(18.52 ±3.47)d, t=-2.613, P=0.011;direct costs:￥(17 029.05±317.28) vs.￥(15 610.64±292.56), t=23.764, P=0.000].When LOSNDI and quality-adjusted life years (QALYs) were estimated as indicators of effectiveness, the incremental cost-effectiveness ratios and incremental cost-utility ratios of group A were ￥844.3 and ￥70 920.5, respectively.Net monetary benefit of group B was more than that of group A.One-way sensitivity analysis showed that parameters had no significant effect on the model.Conclusion When using local per capita gross domestic product as threshold, early postoperative Ala-Gln PN was more economical than perioperative Ala-Gln PN strategy evaluation.

Objective To explore the application of self-made guided wire operating-tray,which is designed for percutaneous tranluminal coronary angioplasty (PTCA),in performing coronary intervention (PCI).Methods A total of 40 patients who were planned to receive PCI were randomly divided into the study group (n=20,using guided wire operating-tray) and the control group (n=20,using traditional method to operate guide wire).Under local anesthesia PCI was carried out in all patients of both groups.The frequency of guide wire winding,the degree of surgical sheet contamination within the operation area,and the comfort and convenience of operating guide wire were assessed,the results were compared between the two groups.Results The frequency of guide wire winding and the degree of surgical sheet contamination within operation area in the study group were remarkably lower than those in the control group (P＜0.05).The comfort and convenience of operating guide wire in the study group was better than that in the control group (P＜0.05).Conclusion In performing PCI,the use of self-made guided wire operating-tray can effectively avoid guide wire winding and reduce the degree of surgical sheet contamination within the operation area.

To compare the clinical effect of phacoemulsification and middle incision extracapsular cataract extraction ( Ml-ECCE) . METHODS: One hundred and eighty - five eyes of phacoemulsification (137 cases) and 185 eyes of 139 cases for Ml-ECCE from January 2011 to May 2013 were involved in this study. And the ratio posterior capsular rupture during surgery, visual acuity, corneal edema, corneal astigmatism and intraocular pressure post operation were followed up.RESULTS: On 1d after surgery, uncorrected visual acuity in the group of Ml-ECCE was better than that of phacoemulsification group, while from 3d; 1 and 3mo after surgery, no significant difference was found from the above two groups. On 1d postoperation, corneal edema ratio in phacoemulsification group ( 45 eyes ) was higher than that in Ml-ECCE group(20 eyes) ( X2=11. 665, P=0. 0006 ) . No significant difference was found for the ratio of posterior capsule rupture during surgery in these two surgical technique groups(X2=0. 094,P=0. 759). On 1wk;1 and 3mo after surgery, significant difference was found for the average of surgical induced corneal astigmatism between two groups (u=6. 661, 6. 880, 4. 187, P = 0. 00, respectively ). During following up, no significant difference was found for the intraocular pressure between two groups ( u=1. 858, 0. 963, 0. 471, 1. 349, 1. 388; P= 0. 063, 0. 335, 0. 638, 0. 177, 0. 165). lntraocular pressure on 1d postoperation in phacoemulsification and Ml-ECCE groups was higher than before operation ( u = 19. 86, 19. 39, P = 0. 00, respectively). And on 1wk; 1 and 3mo postoperation, intraocular pressure in the operated eyes in both groups was lower than before operation for 2~3mmHg.CONCLUSlON: Although phacoemulsification and Ml-ECCE could both get good visual rehabilitation, with similar visual outcome, no significant effect for intraocular pressure, and no severe complications, the latter one owns the advantage that easier maneuver, quicker recovery, and cheaper instruments needed, which is suitable for the hard nuclei cataract in local hospitals. Ml-ECCE is a safe, effective and easy manipulation for local hospital for large batch of cataract surgeries.