Objective To reconfirm the mesangial cell differentiation potential of BMSCs and to study the mechanism of cell induction.Methods Rat BMSCs were isolated and incubated with platelet-derived growth factor(PDGF)-BB to induce the mesangial cell phenotype.At day 7,the cell morphology,mesangial cell-specific markers and cell contraction function were examined.MAPK/ERK kinase inhibitor(PD98059) was added to examine its impact on cell growth and mesangial cells marker expression.Results After cultivation under the differentiation condition for 7 days,the induced cells expressed ?-actin and Desmin,which highly resembled cultured mesangial cells in rat.The induced cells with increasing level of PDGF receptors mRNA expression changed into a wide range of shapes from spindle to stellate and were contracted in response to angiotensin Ⅱ.PD98059 perturbed the induced cells mesangial cell-specific markers mRNA expression.Conclusion BMSCs can differentiate into mesangial-like cells with PDGF-BB induction in vitro.The mechanism involved in the differentiation was mediated through MAPK/ERK1/2 signal transduction pathway.

Background Cyclosporine A (CsA) is an effective drug to prevent rejection response after penetrating keratoplasty (PKP).Appropriate dosage forms and right administrating route is very important for improving the bioavailability of CsA.Objective This study was to investigate the preventing effect of CsA microspheres via subconjunctive and anterior chamber injection and CsA eye drops on immune rejection after PKP.Methods Sixty eyes of 60 clean adult New Zealand white rabbits served as receipts,and 60 eyes of 30 clean adult pigmented rabbits were used as the donors.The receipts were randomized into the blank control group (only PKP group),subconjunctival CsA injection group,subconjunctival vector injection group,anterior chamber CsA injection group,anterior chamber vector injection group,and 1％ CsA eye drops group.PKP was performed on the all rabbits,and then the CsA microsphere (0.1 ml,12 g/L) or blank microsphere (0.1 ml) were respectively administered in the corresponding groups.The corneal grafts were examined by slit lamp microscope regularly,and rejection index (RI)was calculated based on the corneal opacity,edema and neovascularization.The intraocular pressure (IOP) of operative eyes was measured by Tono-pen tonometer before operation,3 days,1 week,2 weeks,3 weeks,1 month,2 months and 3 months after operation,respectively.Histopathological examination on corneal grafts was performed 1month and 3 months after operation.Results The IOP of all the rabbits lowed after operation,but there was no statistical difference in different time points and various groups (Ftime =29.210,P =0.000; Fgroup =0.254,P =0.938).The grafts of the blank control group,subconjunctival vector injection group and the anterior chamber vector injection group showed varied degrees of corneal opacity and neovascularization 2-3 weeks after operation,and the degree of graft opacity was aggravated obviously in the fourth week,with the RI 8.60±1.52,8.60±0.55 and 8.80±0.84 individually.Neovascularization of the grafts in the subconjunctival CsA injection group,anterior chamber CsA injection group,and 1％ CsA eye drops group was found 3 weeks after operation,and the RI were 4.40±0.89,3.20±0.84 and 3.00±0.71,showing a significantly lower than that of the control groups (P＜0.05).Mild inflammatory response of the grafts was seen in the anterior chamber CsA injection group,but it was lightened over time.The histopathological examination revealed obvious thickening of corneal grafts,neovascularization and infiltration of lots of inflammatory cells in the blank control group,subconjunctival vector injection group and the anterior chamber vector injection group.However,only slight new blood vessels and inflammatory response were seen on the subconjunctival CsA injection group,anterior chamber CsA injection group,and 1％ CsA eye drops group.Conclusions Administration of CsA in different methods can prevent the immune rejection after PKP in rabbit eyes,the effect of giving drugs via anterior chamber injection is better than that via subconjunctive pathyway.

Proper timing of repair is one of key factors predicting long-term prognosis of iatrogenic biliary injury.Local inflammation is proved related to long-term biliary stricture.This article introduces pathological procedure of biliary injury based on pathophysiological mechanism and animal model rescarch of wound healing,and how to increase intraoperative repair rate based on the clinical evidences.The preoperative active inflammation control and systemic management could create necessary conditions for the the subsequent early repair.At the same time,authors suggest to set individual strategy regarding timing of repair.Delayed repair is recommended for combined vascular injury or severe biliary injury with terrible contamination.

Objective To explore the changes of the levels of interleukin-17(IL-17),matrix metalloproteinases-9(MMP-9) and immunoglobulin E(IgE) in serum of asthmatic children and analyzing the correlation.Methods Twenty-eight asthmatic children and 14 healthy children were collected to determin the levels of IL-17,MMP-9 by sandwich enzyme linked immunosorbent assay,and the correlation of IL-17,MMP-9 and IgE in serum were analyzed.Results The levels of IL-17,MMP-9 and total IgE in serum of asthmatic children were significantly higher than that of control group(Pa0.05).Conclusions The levels of IL-17 and MMP-9 increase in asthmatic airway inflammation.IL-17 and MMP-9 play an important role in asthmatic airway inflammation and airway rebuilding.

Background The select of supporter is critical for the construction of tissue engineering cornea.Many carrier carl be utilized in the construction of tissue engineering cornea,but de-cellular corneal matrix is known to be one of optimal supporters.Objective Present study was to investigate the characteristics of de-cellular corneal matrix of porcine of structure and biocompatibility for rabbit cornea stroma and limbal epithelial ceHs. Methods The porcine cornea was prepared as de-cellular corneal matrix of porcine by the application of detergent enzyme combined process.The corneal epithelial cells,keratocyte and endothelial cells of porcine were removed completely and stored in -20℃ refrigerator after sterilization.The morphology of de-cellular corneal matrix of porcine was examined by hematoxylin-eosin staining under the light microscope.The structure characteristics of de.cellular corneal matrix of porcine under the scan electron microscope,and its physics features were investigated by the evaluation of water content,strength,expansion and transparency.The de-cellular corneal matrix of porcine were implanted to cornea stroma of rabbit and co-cultured with rabbit corneal epithelial cells for 4 weeks in vitro to assess the keracyte compatibility. Results The epithelial cells,keratocyte and endothelial cells of porcine were removed completely by trypsogen digestion.The collagen fibril network and collagen plates paralleled to corneal surface under the light microscope.The water content,strength,expansion。Ratio of light transparency of de-cellular corneal matrix of porcine were similar to normal porcine cornea.After implantation of de.cellular comeal matrix of porcine into rabbits corneal stroma,the edema of tissue was found in one week,and edema disappeared on two weeks and became clear on four weeks after surgery.The de-cellular eorneal matrix attached to rabbit cornea stroma well.No inflammatory eell and new vessel were found after surgery.The co-cultured rabbit corneal epithelial cells differentiated and proliferated on the surfaee of de-cellular corneal matrix and showed positive response for CK3.No statistically significant differences were found in the water content,strength,expansion of de-cellular cornea matrix of porcine among the normal,before dehydration,2 and 4 hours after dehydration cornea matrix(P＞0.05).However,the transparency was much better in the corneal matrix with 2-hour,4-hour dehydration in comparison with non-dehydration one(P＜0.05). Conclusion The structure features of de-cellular cornea matrix of porcine are similar to normal porcine cornea.Good biocompatibility is proved for xenogenesis of rabbit cornea.

NSP4, as the diarrhea-related protein of rotavirus, is becoming an attractive candidate for vaccine development. To compare the immunogenicity of NSP4 from different genetic groups, we constructed eukaryotic expression plasmids comprising the NSP4 genes from four different genetic types using the pCI vector. The recombinant vectors were designated as pCI-97B6, pCI-97S36, pCI-97S34 and pCI-97SZ8, respectively. Following the conformation of the transient expression of the constructs in 293 cells, the plasmids were respectively subjected to the 5 round i. m. inoculation of BALB/c mice. The specific antibodies against NSP4 as well as the IgG1/IgG2a subclasses of immunoglobulin in mice sera were examined with indirect ELJSA after each immunization. The results showed that the immunization of plasmids expression NSP4s could elicit not only humoral but also cellular immunity, but the humoral immune response is dominant. There is a difference of immunogenecity among the NSP4 of different genetic type. Further studies were needed to focus on the relationship between the immunogenicity and protection effect.

Objective To analyze and compare the clinical characteristics of primary sclerosing cholangitis (PSC) with IgG4-related sclerosing cholangitis (IgG4-SC).Methods The clinical data of 32 PSC patients and 72 IgG4-SC patients who were hospitalized in Peking Union Medical College Hospital (PUMCH) from January 2004 to December 2014 were retrospectively analyzed.Results Of the 32 PSC patients,there were 16 male and 16 female.Of the 72 IgG4-SC patients,there were 61 male and 11 female,(ratio =5.5∶ 1).The average ages were 44.9 (11 ～ 77) and 59.8 (28 ～ 83) years,respectively (P ＜0.05).The most common symptoms of PSC and IgG4-SC were abdominal pain and jaundice,and the incidences of abdominal pain and jaundice were 50.0％ and 68.1％,78.1％ and 81.9％,respectively.The serum IgG4 level of the IgG4-SC patients was significantly higher than the PSC patients (P ＜ 0.05).The total protein in serum of the IgG4-SC patients was higher than the PSC patients (P ＜ 0.05).The rate of bile duct wall thickening as detected on endoscopic ultrasonography (EUS) was higher than by abdominal ultrasound and abdominal CT,which were 91.2％,11.5％ and 33.3％,respectively (P ＜0.05).12 PSC patients were followed up for over 2 years,including 2 patients who underwent liver transplantation after failure of conservative treatment,5 patients who died from hepatic failure and infection,and 3 with stable condition.43 IgG4-SC patients were followed up for over 2 years,including 16 patients with relapse.The recurrence rate was 37.2％ (16/43).The more the extrabiliary organs or bile duct segments were involved,the higher was the recurrence rate.Conclusions Both PSC and IgG4-SC are cholestatic diseases,and they have many similarities in clinical and imaging manifestations.However,they still have unique features.IgG4-SC is sensitive to glucocorticoids therapy and has good prognosis.Thus,it is important to differentiate PSC from IgG4-SC.

Objective To investigate the feasibility of 18G trocar for central venous catheterization in adults.Methods Retrospective analyzed the clinical data of 60 patients with central venous catheterization under local anesthesia.These patients were randomly divided into the control group and the observation group.Selected the internal jugular vein as the site of the puncture catheter.The control group was punc-tured by conventional puncture needle while the observation group was punctured by the 18G trocar.Results There was no statistical differ-ence between the two groups in the success rate of final puncture and catheterization(P>0.05).Compared with the control group,the suc-cess rate of guidewire insert was higher,the puncture operation time was shorter,the pain score during puncture operation was lower,and post-operative patient satisfaction was higher in the observation group.The differences of the two groups were statistically significant(P<0.05). Conclusion 18G trocar can perform central venous catheterization successfully.This method has great advantages for awake patients,and it also worked in some cases with difficult wire insertion.

Objective To explore the distribution of arsenic speciafion and to estimate the effect of arsenic on glutathione(GSH)levels in the blood and liver of mice exposed to different concentrations of inorganic AsⅢ through drinking water.Methods Mice drank water containing arsenite at concentrations of iAsⅢ of 0(contr01),25,50,100 ms/L for 6 weeks.Blood and liver were sampled to asses$the levels of inorganic arsenic(iAs),monomethylarsenic acid(MMA),dimethylarsenic acid(DMA)by the method of hydride generation trapping and ultra-hypothermia coupled with atomic absorption spectrometry,and the level of GSH by the method of 5,5'-Dithio-bis (2-Nitrobenzoic acid).Results Leveh of iAs.MMA and DMA in blood and in liver increased along with the increase of iAs concentrations in drinking water.Primary methylated index(PMI)and secondary methylation index (SMI)of liver and blood were significantly higher in exposed groups than those in control group(P＜0.05).SMI of liver in 50 mg/L exposed group[(50.45±2.94)%]was significantly higher than those in 25 mg/L and 100 mg/Lgroups[(41.68±7.09)%and(41.19±8.87)%,respectively],the difference being statistically significant(P＜0.05).The ratio of iAs.MMA and DMA in blood and liver in exposed group were 2:3:5 and 4:3:3,the percentage of level of organic arsenic(MMA+DMA)were 80%and 60%.GSH in blood and liver in exposed group decreased along with iAs concentrations in drinking water and had significant differences compared with those in control group (P＜0.05).However,levels of GSH in liver and blood did not differ significantly between exposed groups and control group(P＞0.05).Conclusions Membolism of iAs in liver is maximized when the iAs concentrations in drinking water increases to a certain level.However,the percentage of arsenic speciation in blood is different from that in liver,suggesting that other organs and tissues may be capable of methylation of inorganic arsenic.The level of GSH in liver and blood in mice is a good mark tO reflect the toxicity of arsenic.

OBJECTIVETo investigate the risk factors related to post-asphyxial multiple organ dysfunction (PA-MOD) in neonates.

METHODSA total of 397 neonates with birth asphyxia were enrolled from January 2009 to December 2010.The patients were divided into PA-MOD group (n=179) and non-PA-MOD group (n=218). The risk factors of PA-MOD were retrospectively studied.

RESULTSMultivariate logistic regression analysis showed that severe asphyxia, fetal distress, abnormal labor, and decreased amniotic fluid were the risk factors for PA-MOD among the neonates. Spearman rank correlation analysis showed that the number of the involved organs increased along with the increase of age at admission (P<0.05) and with the decrease of gestational age and birth weight (P<0.05).

CONCLUSIONSThe efforts should be made to enhance perinatal care for neonates, especially for preterm infants and low-birh-weight infants, to decrease the incidence of MOD.

Asphyxia Neonatorum ; complications ; Female ; Humans ; Infant, Newborn ; Logistic Models ; Male ; Multiple Organ Failure ; etiology ; prevention & control ; Risk Factors