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 and analyze the cause, prevention and treatment for complications in patients with serious hypospadias repaired by one-stage urethroplasty. Methods From 1987 to 2002,275 patients with serious hypospadias were repaired by one-stage urethroplasty, there were 35 cases had complications. The classifications were penoscrotal 148 cases, scrotal 95 cases and perineal 32 cases. The lengths of new urethras were from 3.0 to 8.4 cm, the mean was (4.1?0.7) cm. Thirty-two cases were received endocrinotherapy before urethroplasty. Results The rate of complications was 12.7% for 1～3 years following survey. There were urethral fistulas 24 cases (8.7%),urethral strictures 6 (2.2%), diverticulums 3 (1.1%), chordees 2 (0.7%).The rate of urethral fistulas was the first and urethral stricture was the second, they were higher than those of other complications (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.

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

PurposeTo optimize the imaging parameters of clinical MRI scanner in rat pancreas imaging to improve the image quality and to provide better MRI image quality and more economical research method for imaging study of rat pancreas. Materials and Methods Twenty-four healthy male Wistar rats were randomly divided into the conventional sequence (CS) group, the adjustment sequence (AS) group and the optimization sequence (OS) group, with 8 rats in each group. The rats in the CS group were scanned with conventional parameters using a clinical MRI scanner. The principle of parameter adjustment was: parameters associated with T1WI or T2WI imaging quality (TR, TE, slice thickness, NEX, FOV and matrix) was set with four changes, and only one of the six parameters was changed in each scan, image quality was evaluated by two senior radiologists, the parameter corresponded the best image quality evaluated consistently by two radiologists were selected as the optimal imaging parameter, all the optimized parameters were set up step by step in this way which formed the imaging parameters in OS group. The pancreatic signal intensity and signal to noise ratio was compared between CS group and OS group after imaging.Results The optimized sequence parameters in clinical MRI scanner were listed below: T1WI sequence (M3D/FSPGR/15): TR 6 ms, TE 2.5 ms, slice thickness 2.0 mm, NEX 8, FOV 7 cm×7 cm, Matrix 120×120; T2WI sequence (FSE-XL/90): TR 4000 ms, TE 71 ms, slice thickness 2.0 mm, NEX 1, FOV 8 cm×8 cm, Matrix 192×160. The pancreatic SI in T1WI and T2WI sequence of the OS group were significantly higher than those in the CS group (t=5.16 and 3.80,P<0.01), while the pancreatic SNR in T1WI and T2WI sequence of the OS group were significantly higher than those in the CS group (t=5.65 and 3.26,P<0.01).Conclusion The optimized parameters can improve the imaging quality of rat pancreas MRI significantly, thus provide a reference for the related experimental study.

Objective To analyze the genomic sequences of Cryptococcus neoformans var grubii strains of two genotypes with different virulence and to screen out the virulence-associated genes. Methods A clinical strain (IFM56800) with the strongest virulence and an environmental strain (IFM56731) with the weakest virulence were screened out for whole genome sequencing analysis. The results of sequencing analy-sis were comprehensively analyzed by using the method of comparative genomics. Genetic variations were ex-tensively screened by using the strategies of non-synonymous single nucleotide polymorphisms ( nsSNPs), nonsense SNPs and the insertions or deletions ( InDels) causing frameshift mutations. The filtered genes were sequenced in 20 experimental strains. The whole RNAs were extracted and then the full-length cDNAs were sequenced by using the rapid amplification of 5′ and 3′ cDNA ends (RACE) method. Results By whole genome sequencing, valid data with high coverage (127 times and 111 times) was obtained in both the environmental strain IFM56731 and the clinical strain IFM56800. The data of InDels and SNPs were statisti-cally analyzed, respectively. Six genes were chosen for further analysis based on the strategies of nonsense SNPs and the InDels causing frameshift mutations. The six genes were amplified and sequenced in all of the experimental strains, three of which were further analyzed with cDNA sequencing. Ultimately, the location and structure of CNAG_01032 gene were determined. The predicted nonsense mutation locus was verified to present in the actual mRNA. Conclusion The strategies of nonsense SNPs and the InDels causing frame-shift mutations showed high-efficiency in screening potential virulence-associated genes. The CNAG_01032 gene was screened out as a novel virulence-associated gene.

Objective To detect the serum levels of tumor necrosis factor-alpha (TNF-α) and interferengamma (IFN-γ) in brucellosis patients and to study the Th1 immune response in acute and chronic patients.Method Serum levels of TNF-alpha and IFN-gamma of 110 brucellosis patients,including 58 acute brucellosis patients and 52 chronic brucellosis patients,were measured by enzyme linked immunosorbent assay (ELISA) from 2014 to 2015 in Zhangjiakou Infectious Disease Hospital.Results The serum levels of TNF-alpha and IFN-gamma of 58 acute brucellosis patients were (38.2± 3.6) pg/L and (31.3 ± 3.7) ng/L,respectively;the serum levels of TNF-alpha and IFN-gamma of 52 chronic brucellosis patients were (12.4 ± 2.6) pg/L and (8.8 ± 3.4) ng/L,respectively.The differences were statistically significant between acute and chronic patients (t =43.216,33.809,all P ＜ 0.05).The early cure rate,early base cure rate,improvement rate and inefficiency rate were 36.2％ (21/58),32.7％ (19/58),25.9％ (15/58)and 5.2％ (3/58),respectively in acute patients.Inversely,they were 17.3％ (9/52),13.5％ (7/52),15.4％ (8/52)and 53.8％ (28/52),respectively in chronic patients.The therapeutic effect was better in acutepatients than chronic patients (x2 =4.937,5.657,all P ＜ 0.05).Conclusion It seems that acute brucellosis patients have a higher serum levels of TNF-alpha and IFN-gamma and a better prognosis due to effective Th1 immune response,and chronic brucellosis patients are associated with poor outcome due to deficiency of Th1 immune response.

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.

Objective To investigate the genetic relation between Cryptococcus neoformans var.the clinical strains in MLMT - 13 genotype and the environmental strains in MLMT - 36 genotype. Methods Multilocus microsatellite typing (MLMT) method was applied for the genotype analysis in our study.Through this method, we recognized two genotypes that distinguish a majority of clinical and environmental strains. In order to compare virulence between the two types, we chose to infect BALB/c mice (6 weeks,female) with 9 MLMT-13 strains and 10 MLMT-36 strains intravenously. Results Forty( 17 clinical and 23 environmental isolates) were analyzed. Of 17 clinical strains, 9 belonged to a major type of MLMT-13 (52.9%). They were mainly isolated from clinical specimens. About 43.5% of strains from the environment belong to a major type of MLMT-36, which are indigenous to environments and which were not isolated from clinical samples. The mortality rate and pathological changes of the above mice were observed during two months after injection. The results showed that the mortality rate of mice infected with MLMT-13 strains was 100%, while the mortality rate with MLMT-36 strains was 7. 5%. The pathological sections showed that lesions of MLMT-13 infected mice appeared in the brain, lungs, liver and kidneys, while the lesions of MLMT-36 infected mice only appeared in the brain. Most brains of MLMT-13 infected mice were distorted,and both the number and size of lesions in such brains were much larger than those of MLMT-36 infected mice. Conclusion Our study illustrated the virulent difference between MLMT-13 and MLMT-36, which are isolated from patients and environment respectively. The results inferred that some genetic changes, such ss microsatellite repeats, might occur between environmental and clinical isolates through their environmental adaptation progress.