AIM:To investigate apoptosis and the expression of death receptors of TRAIL, TNF and Fas on hepatic veno-endotheliocyte ED25 cell strain induced by dengue virus type 2(DV2).METHODS: Flow cytometric analysis was used to detect the number of apoptotic cells and the expression levels of TRAILR1-4 ,TNFR1-2,Fas on ED25 cells before/after DV2 infection. RESULTS: The numbers of apoptotic cells of ED25 increased after DV2 infection, there were only about 5.7%?1.2% of apoptotic cells before virus infection while there were approximately 27.3%?1.6% of apoptotic cells after virus infection. At the same time the expression level of Fas also increased, before virus infection about 44.3%?2.2% of ED25 cells expressed Fas while 63.0%?2.3% of ED25 cells expressed Fas after virus infection. CONCLUSION: DV2 infection can induce apoptosis of ED25 cells, and it suggests strongly that Fas/FasL may be involved in the apoptotic signal transduction.

The integrated courses in pathophysiology have been implemented for clinical medicine in Nanfang Medical University with experimental contrast method since 2006 to compare the advantages of the teaching effect of the integrated courses with traditional learning in pathophysiology with the questionnaire. The results have showed that the students on the integrated track are more positive in the knowledge learning,problem analysis and solving as well as the subsequent clinical knowledge learning.

In order to investigate the dynamic changes in nitric oxide synthase (NOS) activity in organs during traumatic shock and to evaluate the effects of inhibitor of NOS and L Arginine (L Arg) on traumatic shock, rat model of traumatic shock was established, and the constitutive NOS and inducible NOS in the heart, small intestine, liver, lung , spleen were measured 0 5h, 3h, 5h after traumatic shock. Followed by intravenous injection with aminoguanidine (AG ), N G nitro L arginine methyl ester (L NAME), or L Arg, NOS were again measured, and the survival time and 24h survival rate were recorded. It was found that in normal rats cNOS was expressed in all organs, but only a weak iNOS expression in lung and liver. Half an hour after shock, cNOS was elevated in almost all organs in various degrees but there was no change in iNOS. 3h after shock, an increase in iNOS activity and a decrease in cNOS were observed. The iNOS was synthesized in large quantity 5h after shock. AG and L Arg markedly prolonged the survival time but L NAME did not in shock rats. AG inhibited the activity of iNOS but promoted the synthesis of cNOS, and both NOS were inhibited in L NAME group, but not in L Arg group. The results that iNOS was synthesized in large quantity only at later period of traumatic shock. To treat shock, the inhibitors of NOS and L Arg should be given according to the degree of shock, the dosage of drugs, and the time of administration

Objective To investigate the alternation and significance of P-selectin in plasma during traumatic shock.Methods 24 rats were randomly assigned to three groups;control group, shock group and L-Arg group. The rats were subjected to induce traumatic shock ,and to determine the plasma P-selectin with ELISA after shock 30 min and 4 hour.Results P-selectin was expression in plasma of normal rats. 30 min after shock, P-selectin in sera was higher than in the control nontraumatized,but no statistical significance;4h after shock, the P-selectin was significantly increased. The P-selectin of L-Arg group in L-Arg group was obviously decreased than those of shock group.Conclusions Traumatic shock could induce P-selectin hyperexpression.It is related to remarkable microcirculatory perturbation and endothelial dysfunction during shock.L-Arg maybe decrease the expression of P-selectin through promoting the synthesis of the endothelial NO.

Objective To investigate the surgical treatment of male gynecomastia. Methods The clinical data of 116 cases(156 sites) of male gynecomastia were treated by using micro incision of areola with tumescent anesthesia in the past 4 years were reviewed. Results In this series, 50 to 150 grams mammary gland tissue were resected from the breast. There were no severe postoperative complications,such as nipple or areola necrosis and apparent scar . Follow up was done for 12 to 36 months, the scars were obscure, and the configuration of the breasts were good. Conclusions The operation for male hynecomastia via micro incision of areola with tumescent anesthesia is a good, reliable and convenient technique. It is easily performed with milder injury,less bleeding, and quick recovery.

BACKGROUND:There are many preliminary studies on the survival, metaptosis, and correlation characteristics of human amniotic epithelial cel s after transplanted into the animals, but there are no reports on the quantitative analysis of the transplantation effect. OBJECTIVE:To make quantitative analysis on serum biochemical function of liver and the expression of human albumin in mice received passaged human amniotic epithelial cel s transplantation in spleen. METHODS:Forty nude mice were randomly divided into four groups (n=10 in each group):hepatectomy+cel transplantation 2 weeks group, hepatectomy+cel transplantation 4 weeks group, hepatectomy+normal saline group (treated with partial hepatectomy) and hepatectomy+cel transplantation group (transplanted with 0.2 mL passaged human amniotic epithelial cel s with 5×106 under spleen, and the blood were col ected at 2 and 4 weeks after transplantation). The mice in the hepatectomy+normal saline group were treated with splenic injection of 0.2 mL normal saline;the cel transplantation group did not receive hepatectomy, and transplanted with 0.2 mL passaged human amniotic epithelial cel s with 5×106 under spleen. The histological and morphological changes of the liver and spleen in each group as wel as the expressions of serum alanine aminotransferase, aspartate aminotransferase and human serum albumin in each group were detected, and the quantitative analysis of human serum albumin expression was performed. RESULTS AND CONCLUSION: There was no obvious morphological change after human amniotic epithelial cel s transplanted into the acute liver injury mice for 4 weeks, but specific cel s could be detected by histological method. The serum levels of alanine aminotransferase, aspartate aminotransferase and human serum albumin were improved obviously, and the human albumin could be detected in serum, the level of human albumin at 4 weeks after transplantation was significantly increased than 2 weeks after transplantation. Human amniotic epithelial cel s can survive for more than 4 weeks after transplanted into the liver injury mice, and can stil express partial characteristics and functions of hepatocyte-like cel s, improve the liver function, thus treating acute liver injury.

BACKGROUND:Human amniotic epithelial cells(AECs)are easy to obtain and can function as ideal seed cells for cell transplantation and tissue repair.Currently,marking and tracing of human AECs remains rarely reported.OBJECTIVE:To explore the efficiency of adeno-associated virus(AAV)vector-medicated green fluorescent protein(GFP)on in vitro cultured human AECs transfection.METHODS:Human amnion samples were harvested and trypsinized to isolate human AECs.The AECs were transfected with AAV-GFP,and the transfection efficiency was detected.RESULTS AND CONCLUSION:Human AECs were successfully primary cultured and passaged in vitro.AAV-GFP-transfected AECs stably and highly expressed GFP,with a transfection efficiency of 58%.

Objective To evaluate the in vitro differentiation of human amniotic epithelial cells (hAECs ) into hepatocyte-like cells. Methods Combined approach of dexamethasone, HGF, IGF and other cytokines were used to induce the differentiation of hAECs into hepatocyte-like cells. The induction lasted 2 weeks. During the induction, the expression of albumin ALB, CYP1A1, CYP1A2, IGFR, c-met and key functional genes related to liver cells as well as transcription factors HNF3, HNF4 and C/EBPa were monitored by RT-PCR. Time dependent changes of the surface marker colony ALB, AFP and CK18 were analyzed by cell flow cytometry. Results After the 2 week induction, the expressions of liver hepatocyte-like cell functional genes such as albumin, CYP1A1, CYP1A2, c-met, and transcription factors such as HNF3, HNF4, C/EBPa and HNF1 were observed. Six days after the induction, hAECs mainly were stained AFP+, and the positive rate was (15.1±2.1)%. While 10 days after the induction, part of the hAECs showed AFP+/ALB+ (6.5±1.4)%; and on 14th day, hAECs only showed ALB+, and the rate was (13.9±2.3)%. ALB+ cell increase indicated a gradual functional maturation from the hAECs to hepatocyte-like cells. Similaritly, the number of CK18+ cells in the whole population was also increased: On 10th day, the rate was (16.1±1.2)%; on 14th day, that was (21.3±4.6)%, which proved the above hypothesis of the trandifferentiation. By extending the induction time, the expression of functional genes increased gradually, and a maturing process of hAECs was detected by cell surface markers. Conclusion The differentiation of hAECs induced in vitro has the characteristics of hepatocyte-like cells.

Objective To investigate the high risk factors of persistent pulmonary hypertension (PPHN) in preterm and full-term neonates.Methods Fifty-six newborns with PPHN were divided into preterm neonates group and the full-term neonates group according to their gestational ages.The relationship between PPHN and high risk factors such as the primary diseases,mother's complications during pregnancy,and the prognosis were analyzed retrospectively in each group.A retrospective review of lung samples of 3 neonates who died of respiratory failure were carried out.Results Thirty-one (55.36％) out of 56 neonates with PPHN including 27 preterm and 29 full-term newborns had parenchyma diseases.The first and the second primary diseases were meconium aspiration syndrome (MAS,9/29 cases,31.03％) and congenital heart diseases (8/29 cases,27.59％) in the full-term group,and respiratory distress syndrome (RDS,16/27 cases,59.26％) and infection (8/27 cases,29.63％) in preterm group,respectively.There were 15 cases (55.56％) whose mothers had complications during pregnancy among the 27 preterm neonates,and 8 ca ses (27.59％) among the 29 full-term neonates.The morbidity of mothers' complications during pregnancy in preterm neonates was higher than that in full-term neonates (x2 =4.519,P =0.034).The mortality rate was 33.33％ (9/27cases) in the preterm neonates group while 27.59％ (8/29 cases) in the full-term neonates group.There was no significant difference in mortality rate between each group (x2 =0.218,P =0.640).The primary diseases were RDS (9/9cases,100％) and MAS (4/8 cases,50％) in preterm and full-term neonates who died of PPHN,respectively.One case of pulmonary hypertensive vascular remodeling,1 case of alveolar capillary dysplasia (ACD) and 1 case of interstitial pneumonia were found in 3 lung samples of neonates died of PPHN.Conclusions The ratio of preterm increased in neonates with PPHN.RDS and infection were the first two primary diseases in preterm neonates with PPHN.MAS was the first primary disease in full-term neonates with PPHN.Primary pulmonary hypertension should be considered for neonates with severe PPHN.

Objective To discuss the surgical treatment for huge hepatoblastoma in children,and the technique of hepatectomy without blockade of the blood supply to the remained liver lobes.Methods We reviewed 12 cases of huge hepatoblastorna who had been operated from July 2001 to January 2009 in our hospital.The mean age of the children was 3.2 years(range,11 months to 12 years).The diameter of the tumor was from 10 to 23 cm.3～7 cycles of chemotherapy was routinely administrated before operation.When the tumor reduced to a certain size that radical resection could be performed safely,regular hepatectomy was conducted.Hepatoblastoma resection without blocking the blood supply to the remained liver lobes was performed in every patient.Results The operations were successfully accomplished in all the 12 children.5 cases received right trihepatectomy (segment Ⅳ,Ⅴ～Ⅷ),4 cases received right hemihepatectomy(segment Ⅴ～Ⅷ),and the other 3 cases received Ⅳ,Ⅶ,Ⅷ segmentectomy,right Ⅴ,Ⅵ segmentectomy,and left hemihepatectomy respectively.The intraoperative hemodynamic parameters were stable,and there was no perioperative mortality.Postoperative chemotherapy wag routinely administrated.The follow-up period varied from 2 to 92 months.11 children survived and were disease free,among those 6 children have survived for more than 5 years.One child had brain and lung metastasis 5 months post operation,and died 7 months post operation. Conclusion Preoperative chemotherapy administrated to children with huge hepatoblastoma can reduce the tumor size and render tumor reseetable.Hepatoblastoma resection without blocking the blood supply to the remained liver lobes is a safe and feasible surgical technique.