Objective To investigate the functional role of mur in e hepatocyte growth factor (HGF) in inducing in vitro the mouse embryonic st em cells (ESC) to differentiate unidirectionally into hepatocyte. Method s Both spontaneous and HGF induced differentiations of the primordial c ell elusters formed by suspended culture in vitro of ESC were daily observed with microscope for 5-7 days. After 4 weeks of culture, the cells were stained with HE and glycogen staining, their morphology were observed, The synthesis of urea nitrogen and triglycerides in the culture medium, the expression of myocard ial MHC, albumin, AFP and CK18, and the indocyanine green (ICG) staining were a lso detected. Results It was hard to control the spontaneous di fferentiation of ESC. HGF could promote the differentiation of ESC into endoderm , and more likely into myocardium. HGF could also induce the expression o f albumin, AFP and CK18, and positive staining of ICG and FDA. Conclusio ns HGF may induce the differentiation of ESC into hepatocyte, but the f unctional role is limited. It implies that a comprehensive effort of multiple fa ctors might be needed in inducing the hepatocyte differentiation.

Objective To study the expression of EI-1 in myocardium during cerebral ischemia and reper-fusion, and to investigate the mechanism of cerebral cardiac syndrome. Method Two hundred and eight SD rats weighting 220～250 gram, were divided into three groups: sham control group (n=48), cerebral ischemia group (n=80), cerebral ischemia/reperfusion group (n=80). The area of cerebral ischemia, and the concentration of sennn ET-1 and CK-MB, and the content of myocardial ET-1 were determined in 0,6, 12,24,48,72 h after cerebral ischemia and reperfusion, and were analyzed by t-test or F-test. Results Cerebral necrosis area was ob-served at 6 h after cerebral ischemia in cerebral ischemia group, and peaked at 12 h (P>0.05). The concentra-tion of CK-MB increased gradually after cerebral ischmia, peaked at 12 h (P<0.05), and then gradually de-creased. The serum concentration of ET-1 peaked at 6 h and then gradually decreased. The content of ET-1 in my-ocardium began to increase at 6 h after cerebral ischemia, and peaked at 12 h (P<0.05). In cerebral ischemia/reperfusion group, all of cerebral necrosis size, CK-MB concentration and myocardial ET-1 concent paelced at 12 h and then gradually decreased (P<0.05). Change of ET-1 concentration in blood was similar to that in cerebral ischemia group. Compared with cerebral ischemia group, the size of cerebral necrosis reduced obviously at 24 h,48 h,72 h in cerebral ischemia/reperfusion group (P<0.05). The concentration of CK-MB in cerebral ischemia/ reperfusion group was higher than that in cerebral isehemia group (P<0.05). The peak time of myocardial ET-1 was shatened in cerebral ischemia/reperfusion group. The change of serum ET-1 was not different between two groups. Conclusions Large area of cerebral ischemia, might cause myocardial injury. ET-1 is involved in the course of myocardial injury following cerebral ischemia. Though cerebral reperfusion can protect brain,but it make myocardial injury more serious,and ET-1 might participate in this course.

Objective To explore the mechanism by which recombinant human growth hormone (rhGH) protects liver function and alleviates portal hypertension in rats with liver cirrhosis. Methods Male S.D. rats with thioacetamide-induced liver cirrhosis were randomly assigned to receive separately normal saline (NS, 0.5 ml) or rhGH(333 ng/kg body weight) daily by subcutaneous injection for up to 7 days. After the respective treatments, changes of GH-binding capacity (R T), GHRmRNA, relative content of collagen (RCC), malon-dialdehyde (MDA), superoxide dismutase (SOD) in liver tissue, serum albumin and ALT and portal vein pressure (PVP) were examined. Results R T (fmol/mg protein) of GHR was respectively 31?4, 40?7(P

Objective To evaluate the influence of the storing time of venous blood samples on the differential count of WBC by Sysmex XE-2100 hematology analyzer. Methods At room temperature, the precision of the differential count of WBC by Sysmex XE-2100 hematology analyzer were tested. 38 samples were taken the differential count of WBC by Sysmex XE-2100 hematology analyzer after stored for 0, 2, 4, 8, 24 and 48 h. Differential count of WBC was also taken under microscope for comparison. Results The precision of differential count of WBC by Sysmex XE-2100for all the samples was in the allowable range. The correlation coefficient of the differential count of WBC by two methods for neutrophils, lymphocytes, monocytes, eosinophils and basophils were 0.9859, 0.9775, 0.8053, 0.8695and 0.5243 (P<0.01). There was insignificant difference in the test at 8h, very significant difference of MONO and EOS at 48 h. Differences of EOS, MONO between two methods were significant increased at 8 h. Conclusion At room temperature, the differential count of WBC of venous blood samples by Sysmex XE-2100 hematology analyzer should finish within 8 h.

Objective To evaluate the identification and counting effeciency of nucleated red blood cells by Sysmex XE-2100 Hematology Analyzer. Methods Accurancy: nucleated red blood cells were counted from 38 specimens by Sysmex XE-2100 Hematology Analyzer and compared with the number counted under microscopy. Precision: 3 specimens with different values were counted for the nucleated red blood cells 10 times by Sysmex XE-2100 Hematology Analyzer and compared with the number counted under microscopy. The CV(% ) value was estimated.Results There was insignificant difference between the results obtained from Sysmex XE-2100 Hematology Analyzer and those under microscopy. In the T-test, P >0.05, r=0.9893(P< 0.01).CV(% ) were 8.1% and 15.8% . It means that the Sysmex XE-2100 is more precise in analyzing the nucleated red blood cells than that under microscopy. Conclusion The nucleated red blood cells count by Sysmex XE-2100 is accurate and fast to obtain clinical data.

ObjectiveTo investigate the unfavorable factors of intestinal mucosa repair after the intestinal epithelial injury in vivo in a mouse model of sepsis. MethodsThe method of cecal ligature and puncture (CLP) was used to induce sepsis and then the intestinal mucosa damage, epithelial cell apoptosis and the number of transformed goblet cells were observed, and the concentrations of serum TNF-αt, IL-1 and TGF-β1 and TFF3 ( trefoil factor 3) in small intestinal mucosa were determined. All above various laboratory examinations were made by different assays including H-E staining, western blot, ELISA and immunohistochemistry respectively. The experimental mice were divided into sepsis group and sham operation control group. The mice with sepsis were separately sacrificed 6 hours ( n = 7 ), 24 hours ( n = 7) and 48 hours ( n = 7) after CLP. Results In septic mice group, the injured intestinal mucosa was found 6 hours after CLP. The damage scores in mice 24 h and 48 h after CLP were higher than those 6 h after CLP, but there was no significant difference between those 24 h and 48 h after CLP. Moreover, a few goblet cells or other epithelial cells adjacent to the injured surface migrated onto the wound to cover the denuded area. The number of goblet cells was substantially decreased in mice of sepsis group 6 hours after CLP compared with sham operation control group. Compared with sham operation control group, levels of IL-1 and TNF-α significantly increased 3-4 times in mice of sepsis group at all intervals, and the phosphorylated caspase-3 increased 4 times. Although TFF3 assayed by using Western blot showed modest increase 6 h after CLP and it declined 24 h and 48 h later. A similar change was found in TGF-β1, it modestly increased 6h after CLP, but it didn't elevate 24 h and 48 h later. ConclusionsSevere sepsis keeps on the inflammatory reaction and epithelial cell apoptosis, preventing the repair of intestinal mucosa from injury.

Aim To investigate the effect of insulin glargine and human insulin on proliferation of a human breast cancer cell line MDA-MB-231 and the role of ERK in the process.Methods MDA-MB-231 cells were incubated with insulin glargine and human insulin at different concentrations and for different time courses.A specific ERK1/2 inhibitor,PD98059,was used either alone or in combination with insulin glargine or human insulin to test the involvement of ERK pathway in cell growth.Cell proliferation was evaluated using cell counting kit-8 reagents.Cell cycle distribution was analyzed by flow cytometry.Results Both insulin glargine and human insulin dose-dependently enhanced MDA-MB-231 cell proliferation at the concentrations from 1 to 100 IU?L-1 after treatment for 96 h.At the concentration of 10 IU?L-1,both drugs promoted cell growth at 48,72,and 96 h.The percentage of S+G2/M cells was significantly increased in both insulin glargine and human insulin treated groups as compared to untreated controls.No significant difference was observed between insulin glargine and human insulin in their effects on cell proliferation and cell cycle distribution.Cell proliferation was significantly inhibited by PD98059.However,in the presence of PD98059,both drugs still promoted cell proliferation significantly as compared to untreated controls.Conclusions Insulin galrgine and human insulin similarly promote proliferation of MDA-MB-231 cells independent of ERK activation.

AIM: To study the differentiation of rat bone mesenchymal stem cells (MSCs) into cardiomyocytes in vitro. METHODS: MSCs were isolated and purified from the bone marrow of rats by density gradient centrifugation and adhering to the plastic culture. The third passage MSCs were treated by 5-azacytidine (5-aza). The induced cells were evaluated by immunocytochemistry staining and RT-PCR analysis. RESULTS: After being induced by 5-aza, some MSCs became bigger and longer. The connection of the cells were formed on day 14.The direction of the cells arraying was similar gradually. The induced cells were stained positively for desmin, ?-actin and troponin I. RT-PCR showed that these cells expressed ? myosin heavy chain. CONCLUSION: 5-aza can induce MSCs to differentiate into cardiomyocytes in vitro.

Objective:To analyze the clinical features, differential diagnosis, treatment and prognosis of complex lymphatic malformations.Methods:The clinical data of patients with complex lymphatic malformation were retrospectively analyzed from April 2010 to April 2022 in the Multidisciplinary Outpatient Department of the Vascular Disease Team of West China Hospital, Sichuan University. All patients were diagnosed with complex lymphatic malformation after consultation with multidisciplinary experts in pediatric surgery, radiology, plastic surgery, pathology, rehabilitation and other departments. The clinical manifestations, blood routine, coagulation function, magnetic resonance imaging and treatment methods of the patients were analyzed. According to the follow-up and disease results, the patients were divided into improvement, stability, progress and death.Results:A total of 18 patients with complex lymphatic malformations were included in the study, including 6 males and 12 females. The age of first diagnosis ranged from 1 month to 29 years old, and the median age was 2.5 years old. Patients were followed up and treated for 0.4 to 12.0 years, with an average follow-up of 3.5 years. Ten patients had pleural and pericardial effusion; 15 patients had visceral involvement which showed multifocal changes in imaging examinations; 9 cases were accompanied by bone destruction, which in Gorham-Stout disease patients broke through the cortex while in generalized lymphatic anomalies it did not; 14 patients had various degrees of coagulation abnormalities, of which 8 patients with severe coagulation dysfunction were all diagnosed as kaposiform lymphangiomatosis. Of the 18 patients, one kaposiform lymphangiomatosis patient died; six patients progressed; eight patients were stable; and three patients improved.Conclusion:The clinical characteristics of patients with complex lymphatic malformations are systemic, diverse and complex. The clinical symptoms of patients with diffuse lymphatic malformation accompanied by involvement of bone and multiple internal organs, chest and abdominal effusion, and coagulation dysfunction should be considered as complex lymphatic malformation. However, due to overlapping clinical characteristics of each subtypes, it is difficult to distinguish patients with complex lymphatic malformation, and the curative effect and prognosis are poor. Precision targeted drugs are the future research direction for the treatment of such diseases.

Objective:To analyze the clinical features, differential diagnosis, treatment and prognosis of complex lymphatic malformations.Methods:The clinical data of patients with complex lymphatic malformation were retrospectively analyzed from April 2010 to April 2022 in the Multidisciplinary Outpatient Department of the Vascular Disease Team of West China Hospital, Sichuan University. All patients were diagnosed with complex lymphatic malformation after consultation with multidisciplinary experts in pediatric surgery, radiology, plastic surgery, pathology, rehabilitation and other departments. The clinical manifestations, blood routine, coagulation function, magnetic resonance imaging and treatment methods of the patients were analyzed. According to the follow-up and disease results, the patients were divided into improvement, stability, progress and death.Results:A total of 18 patients with complex lymphatic malformations were included in the study, including 6 males and 12 females. The age of first diagnosis ranged from 1 month to 29 years old, and the median age was 2.5 years old. Patients were followed up and treated for 0.4 to 12.0 years, with an average follow-up of 3.5 years. Ten patients had pleural and pericardial effusion; 15 patients had visceral involvement which showed multifocal changes in imaging examinations; 9 cases were accompanied by bone destruction, which in Gorham-Stout disease patients broke through the cortex while in generalized lymphatic anomalies it did not; 14 patients had various degrees of coagulation abnormalities, of which 8 patients with severe coagulation dysfunction were all diagnosed as kaposiform lymphangiomatosis. Of the 18 patients, one kaposiform lymphangiomatosis patient died; six patients progressed; eight patients were stable; and three patients improved.Conclusion:The clinical characteristics of patients with complex lymphatic malformations are systemic, diverse and complex. The clinical symptoms of patients with diffuse lymphatic malformation accompanied by involvement of bone and multiple internal organs, chest and abdominal effusion, and coagulation dysfunction should be considered as complex lymphatic malformation. However, due to overlapping clinical characteristics of each subtypes, it is difficult to distinguish patients with complex lymphatic malformation, and the curative effect and prognosis are poor. Precision targeted drugs are the future research direction for the treatment of such diseases.