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%.

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.

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.

Jaundice occurs in 19%-40% of the hepatocellular carcinoma (HCC) patients. HCC associated jaundice may be divided into hepatocellular and icteric types in terms of its underlying pathophysiology. The jaundice of icteric type is caused by obstruction of the bile duct through cancer embolus, blood clot, biliary sludge, tumor compression or infiltration. Jaundice and epigastric discomforts are the main clinical manifestations. In the present case, severe acute pancreatitis and acute cholangitis presenting as initial complaints of icteric type HCC were quite rare. A tumor located at the central lobe of the liver and a cancer embolus at the lower part of the common bile duct (CBD) were detected by CT scan. Curative resection of HCC with CBD exploration eradicated both the tumor and the embolus, and no recurrence was found after a 36 month follow-up.

A total of 16 patients with airway stenosis including benign lesion (n=4) and malignant disease (n=12) were treated with argon plasma coagulation (APC) and Z-type covered retrievable metallic stent.L-and I-type stents were placed by guidance of brenchoscope,while Y-type stent was placed by the guidance of both bronchoscope and fluoroscope.Airway stenosis was from (58.8 +9.1)% before APC to (7.5±2.4)% after APC in tracheal,from (67.4±7.4)% to (19.4±4.1)% in left main bronchus,from (69.6±8.9)% to (27.6±5.4)% in right main bronchus.Symptoms of chest distress and breathlessness were improved remarkably,tachypnea indexes were decreased,but Kamofsky performance scope were increased.Twelve stents were successfully installed by the first time,including 9 Y-shape;otherwise,4 stents failed to be installed because the carinal was too wide to insert the Y-shape stent in two patients with lung cancer,finally,L+I type stents were used.Median survival time after successful stenting was 10 months,and mean time was 13 months.APC combined with bifurcated metal stants relieved obstruction and improved quality of life.

SARS coronavirus (SARS-CoV) is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane (M), nucleocapsid (N) and spike 2 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.

OBJECTIVE: To determine the therapeutic effect of laparoscopic splenectomy, perisoph-agogastric devascularization, and endoscopic variceal ligation (EVL) on patients with portal hypertension. METHODS: We randomly divided 105 patients into 3 groups: 40 had endoscopic band ligation (the ligation group), 35 had splenectomy and perisoph-agogastric devascularization (the laparotomy group), and the other 30 had laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation (the combination group). Blood samples were analyzed preoperatively and postoperatively on day 1,3,and 7,including alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),and directed bilirubin(DBIL). The length of stay, blood loss, operation time, anal exhaust time, azygos vein diameter, blood flow velocity and blood flow, recurrence of esophageal varices and rehaemorrhagia were compared. RESULTS: Between the combination group and the laparotomy group, the serum levels of TbIL and Dbil had difference on 1st postoperative day(P<0.05). AST had difference on 7th postoperative day(P<0.05). The length of stay, blood loss, operation time, and anal exhaust time had significant difference(P<0.05). Among the combination group, the laparotomy group and the ligation group, the azygos vein blood flow before and after the treatment, recurrence of esophageal varices and rehaemorrhagia had no difference(P<0.05). CONCLUSION Laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation have less trauma, lower recurrence rate, fewer complications and rapid recovery, and may reduce the azygous vein blood flow. It can be used safely for portal hypertension.
Adult ; Endoscopy ; methods ; Esophageal and Gastric Varices ; complications ; surgery ; Female ; Humans ; Hypertension, Portal ; complications ; surgery ; Laparoscopy ; methods ; Ligation ; methods ; Male ; Middle Aged ; Splenectomy ; methods

Taking Pharm .D .degree education in School of Pharmacy ,University of Pittsburgh as an example ,this pa-per introduced the American pharmacists training system ,degree setting for service-oriented pharmacy students ,training objec-tive ,course design ,teaching staff and other aspects .This paper also reviewed pharmacy education and professional develop-ments for licensed pharmacists in China .Based on the successful experience abroad ,proposals were made to modify the domes-tic pharmacy education .The process of personnel training should be emphasized to improve the quality and capability of licensed pharmacists in China .

A new method for the simulation of the mass distribution of cancellous bone is presented on the basis of finite element analysis (FEA). In this method,the process of bone remodelling is considered as a process of the topology optimization of a corresponding continuum structure. Fabric tensor is used to express the microstructure and the constitutive properties of cancellous bone. The effective volume fraction or the relative density of a point in the design domain is expressed by the invariables of the fabric tensor. A reference strain interval, which is corresponding to the strain dead zone of a bone in biomechanics, is applied to detect the the final topology of the structure. By the present approach, several numerical results are given, i. e., the simulation on the shape of the coronal plane of vertebrae, the predictions of the mass distributions of the two-dimensional and the three-dimensional proximal femurs. The validity and feasibility of this new method are verified by the comparison between the results of the present work and those in the published literatures.
Adaptation, Physiological ; Algorithms ; Biomechanical Phenomena ; Bone Density ; physiology ; Bone Remodeling ; physiology ; Bone and Bones ; physiology ; Computer Simulation ; Femur Head ; physiology ; Finite Element Analysis ; Humans ; Models, Biological ; Stress, Mechanical

BACKGROUND AND OBJECTIVETranscatheter arterial chemotherapy and embolization is the main method in the treatment of lung cancer, but most of the reports do not study individually to small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), hypovascular and hypervascular lung cancer. The pre-embolization perfusion of hemotherapeutics is still being used routinely and seldom report to iodized oil embolization. The article summarized the quality of life after the treatment, clinical efficiency, survival time and complications to evaluate the clinical effect of primary hypervascular NSCLC treated with embolization of emulsion of chemotherapeutics and iodized oil.

METHODSThe study totally analyzed 41 cases which confirmed by pathology and follow up study from January, 2008 to January 2009. The CT scan with IV contrast demonstrates over moderate enhanced lesion which indicate hypervascular. Within the 41 cases, 23 cases of central, 18 cases of peripheral. Suqamous carcinoma 21 cases, adenocarcinoma 15 cases and squamoadenocarcinoma 5 cases. Stage IIIb 34 cases, stage IV 7 cases. Superselective incubation with microcatheter under DSA, to embolize the capillary bed with liquefied iodized oil and the emulsion of pharmorubicin, to embolize the supply artery of the tumor with gelatin foam microparticle. The liquefied iodized oil 5 mL-10 mL, pharmorubicin 10 mg-30 mg. The longest follow up is 12 months and to compare with the references of 2007-2009.

RESULTSThe KPS is widely acclaimed after the treatment (P < 0.05), no complete response (CR), 31 cases of partial response (PR), 7 cases of no change (NC) and 3 cases of progressive disease (PD), the total efficiency (CR+PR) is 75.6%. The clinical efficiency (CR+PR+NC) is 92.68%. 33 cases of total survival tome over 12 months (80.48%), IIIb stage 29/34 (85.29%), IV stage 4/7 (57.14%). 1 case with severe complication-spinal injury.

CONCLUSIONTo treat primary hypervascular NSCLC with simple embolization of emulsion of chemotherapeutics and iodized oil is very useful and can avoid the side effect of chemotherapeutics. The key point to avoid spinal injury and other severe complications is to distinguish the spinal aretery and intratumor AV fistula by using superselective incubation with microcatheter under DSA.

Carcinoma, Non-Small-Cell Lung ; blood supply ; mortality ; therapy ; Chemoembolization, Therapeutic ; Emulsions ; Female ; Humans ; Iodized Oil ; administration & dosage ; Lung Neoplasms ; blood supply ; mortality ; therapy ; Male ; Middle Aged