Objective To investigate the effect and safety of urinary kallidinogenase on acute cerebral infarction.Methods One hundred and sixty-four patients with acute cerebral infarction were randomly divided into two groups:treatment group with urinary kallidinogenase(86 cases)and control group (78 cases).According to Chinese guidelines for prevention and management cerebrovascular disease,two groups were treated with basic therapy,such as antiplatelet,neurologic protection,blood pressure control,and so on.On basis of control group,treatment group Was administrated intravenous injection of urinary kallidinogenase 0.15 PNA U per day for 10 days.The primary efficacy Was evaluated by the National Institutes of Health Stroke Scale(NIHSS)score and Barthel index.Results The score of NIHSS and Barthel index at 15 days after treatment in the treatment group were higher than those in the control group[(6.67±3.02)scores vs(7.42±3.02)scores;75.36±23.56 vs 68.36±22.36,P＜0.05].Urinary kallidinogenase could significantly reduce neurological deficits in big artherosclerosis type by TOAST typing.Conclusion Urinary kallidinogenase may be effective and safe in the treatment of acute cerebral infarction.

Objective To study the results using T-tube and retroperitoneal space drainage to manage perforating injury of the distal common bile duct(PIDC) during common bile duct(CBD) exploration.Methods We retrospectively analyzed the clinical data of 12 patients who were diagnosed to suffer from PIDC during CBD exploration from 2010 to 2012.Result All these 12 patients who received T-tube and retroperitoneal space drainage,gastrointestinal decompression,nutritional support and antibiotics recovered uneventfully.Conclusion Given that the CBD was unobstructed,T-tube and retroperitoneal space drainage was an good treatment for patients with PIDC.

Objective To investigate the existence of epithelial-mesenchymal transition(EMT)and its relation to the expression of HBx protein in hepatocellular carcinoma(HCC).Methods The expression of HBx protein,E-cadherin,β-catenin,N-cadherin and fibronectin were detected by immumnohistochemistry technique in 76 cases of HCC.Results Among 76 HCC samples,the loss of expressions of E-cadherin and β-catenin were 34%(26/76)and 20%(15/76),respectively.The positive expressions of HBx protein,N-cadherin and frbronectin were observed in 68%(52/76),55%(42/76),46%(35/76)of HCC samples,respectively.The loss of expression of E-cadherin significantly correlated with the positive expression of N-cadherin and HBx protein (P＜0.01).The positive expression of N-cadherin and fibronectin significantly correlated with the loss of expression of β-catenin and the positive expression of HBx protein(P＜0.05).Conclusions EMT exists in hepatocellular carcinoma,and the expression of HBx is significantly associated with EMT.

Objective To investigate the clinical significance and mechanism of WW domain containing oxidoreductase (WWOX) gene and p73 gene abnormal expression in acute lymphocytic leukemia (ALL).MethodsCase-control study was used in the research.Forty-eight cases of bone marrows from ALL patients were collected,including 32 cases newly diagnosed,11 cases with complete remission and 5 case with relapse.Thirty-one cases of bone marrows from non-leukemia patients were used as control group.All the samples were collected from First Affiliated Hospital of Guangxi Medical University from July 2010 to July 2011.The doctors punctured patients' bone marrows 3 milliliters from the left of posterior superior iliac spine.Samples were bottled up with EDTA anti-coagulation tube.1 milliliter bone marrow was used to extract genome RNA with purity from 1.8 to 2.0.And then,the level of WWOX and p73 gene transcripts were tested immediately using reverse transcriptase-polymerase chain reaction (RT-PCR).Meanwhile genome DNA was also extracted from the other 2 milliliter bone marrow with purity from 1.7 to 1.9,which was used to detect the promoter methylation of WWOX gene and the first exon methylation of p73 gene by methylation PCR (MS-PCP).x2 test and Fisher's exact test were used to compare tbe methylation status of WWOX and p73 gene.Results In 31 controls,expression of WWOX and p73 gene mRNA was 94.00％.The total expression frequency of WWOX gene mRNA in 48 ALL samples was 48.00％ (23/48),much lower than control (x2 =17.434,P =0.000 ).There was significant difference (x2 =10.471,P =0.001 ) between newly diagnosed cases 34.38％ ( 11/32),complcte remission cases (90.91％,10/11 ) and control.The total expression frequency of p73 gene mRNA in 48 ALL samples was 56.00％ (27/48),much lower than control (x2 =12.697,P =0.000).There was significant difference (P =0.012 ) between newly diagnosed cases 43.75％(14/32) and complete remission cases 90.91％(10/11).It was unmethylation in 31 controls.The total methylation frequency of WWOX gene promoter region in 48 ALL samples was 44.00％(21/48),much lower than control (x2 =18.473,P =0.000).There was significant difference (P =0.012) between newly diagnosed cases 56.25％ (18/32),complete remission cases 9.09％ (1/11 ) and control.The total methylation frequency of p73 gene the first exon region in 48 ALL samples was 35.00％(17/48),much lower than control (x2 =13.990,P =0.000).There was significant difference (P =0.033) between newly diagnosed cases 46.88％ (15/32),complete remission cases 9.09％ ( 1/11 ) and control.There was a negative correlation between the expression of WWOX gene mRNA and its methylation status(r =- 0.678,P =0.000),the same as p73 gene ( r =- 0.577,P =0.000).ConclusionsThe abnormal methylation of WWOX and p73 gene may be the major mechanism of gene silence in ALL,which leads to no expression of WWOX mRNA or p73 mRNA.And the abnormal methylation of WWOX and p73 gene may be relevant with the process of occurrence and development in ALL.It may be an effective and significant to detect methylation status of WWOX gene and p73 gene for the diagnosis and treatment of ALL patients.(Chin J Lab Med,2012,35:820-825)

Objective To study the serum levels of proinsulin(PI),insulin(INS),proamylin(PA) and amylin(AMY) in abnormal glucose metabolism individuals and to explore the relationship between PI and insulin resistance(IR) and between PA and islet beta cell function.Methods Totally 79 subjects who received Oral glucose tolerance test(OGTT) and insulin release test in our department from March to May 2008 were divided into 4 groups according to the results of OGTT,that is,normal group,impaired glucose regulation(IGR) group,T2DM 1[fasting blood-glucose(FBG)15 mmol/L) group.All subjects underwent examination of anthropometry and serum bio-chemical indicators,HOMA-IR and HOMA-B were calculated.Results No significant difference was found in the serum levels of PI or AMY among the 4 groups,but the level of PA,PI/INS and PA/AMY among the 4 groups did have significant differences(P

Objective: To review the clinical effect of early enteral nutrition in patients with carcinoma of esophagus after operation.Methods: Nutrient canal was indwelt through Fresubin(TP) was given through the nasal-intestinal tube in 120 cases with carcinoma of esophagus after operation.Results: The complications such as anastomotic leakage,lung infection and incisional infection in the patients receiving early enteral nutrition were much less than those in control group.No death occurred.The time for bowel function recovery was much shorter.Conclusion: Early enteral nutrition can promote the recovery of gastrointestinal function,improve the nutrition,decrease the occurrence of complications and raise the successful rate of operation in operative patients with esophagus carcinoma.

OBJECTIVE To investigate clinical features and etiology of inhalation pneumonia.METHODS Totally 108 cases of inhalation pneumonia during from Jan 2000 to Dec 2005 were completely surveyed and analyzed. RESULTS There were underlying diseases and susceptible factors, and it was not typical in their clinical signs and symptoms.Totally 177 pathogens were isolated from sputa. There were 96 strains of Gram-negative bacilli (54.2%), 41 strains (23.2%) of Gram-positive cocci, and 40 strains (22.6%) of fungi. The 45 cases (41.7%) were with polyinfections, and 19 cases (17.6%) with double infections.CONCLUSIONS We should enhance diagnosis of inhalation pneumonia, make rational use of antibiotic, and take vigorous precautions against inhalation pneumonia.

Objective To investigate the diagnostic values of serum free fatty acids (FFA) and urine podocalyxin (PCX) in the patients with type 2 diabetic nephropathy(DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) were enrolled,and they were divided into three groups,normal albumin group(NA),microalbuminuria group(MA) and heavy albuminuria group(HA),based on the urinary albumin-creatinine ratio(UACR).In addition,40 healthy persons participated in medical examination were selected as the control group.Serum FFA levels were detected by a biochemical analyzer and urine PCX levels by ELISA.The correlation between them was analyzed by Pearson correlation,and their diagnostic values in type 2 DN were evaluated by the receiver operating characteristic curve (ROC).Results Serum FFA levels in the NA,MA,HA and control groups were (0.61 ± 0.14),(0.81 ± 0.13),(0.95 ± 0.18) and (0.49 ± 0.11) mmol/L,respectively,and urine PCX levels were (1.86 ± 0.45),(4.47 ± 1.48),(6.72 ± 1.40)and(1.38 ±0.24) ng/mL,respectively.The levels of serum FFA and urine PCX in type 2 DM patients were significantly higher than those in the control group(P ＜ 0.05),and increased with the progression of type 2 DM.Pearson correlation analysis showed that serum FFA levels were positively correlated with urine PCX levels(r =0.73,P ＜ 0.05).The sensitivity of serum FFA,urine PCX and combined detection in the diagnosis of type 2 DN were 74.1％,80.6％ and 89.5％,respectively,and the latter significantly higher than the former two (P ＜ 0.05).Conclusion Serum FFA and urine PCX levels increase significantly in diabetic patients with renal injury,and both of them have important clinical values in the diagnosis of type 2 DN.

Objective To cast light on the mechanisms governing anaplastic thyroid carcinoma (ATC) and to evaluate the minichromosomes maintain protein (MCM) 5 and MCM7 expression in human normal thyroid (NT),papillary thyroid carcinoma (PTC),and ATC samples,as well as in primary culture cells.Methods We tested the expression of MCM5,MCM7and PCNA in NT,PTC and ATC by immunohistochemistry.We used Western blot and Northern blot to test the expression of MCM5,and MCM7.ATC cells were transfected with MCM7 siRNA,and Western blot was used to examine the change of the expression of MCM7.Results In ATC samples,MCM5 had high expression in 65％ patients,and MCM7 had high expression in 73％ patients.The expression of MCM5 and MCM7 in NT and PTC samples were very low which can be ignored.In ATC cells,high MCM5 and MCM7 expression was paralleled by high levels of MCM2 and MCM6.Inhibition of MCM7 protein levels by small inhibitory duplex RNAs could reduce the rate of DNA synthesis in ATC cells.Conclusion MCM protein expression is upregulated in ATC and leads to excessive proliferation of ATC.

BACKGROUND: It is important to study the methods of culturing bone marrow-derived mesenchymal stem cells (MSCs) to obtain a great amount of high purity MSCs for applying ocular tissues constructed by tissue engineering technique to treat eye diseases.OBJECTTVE: To separate and culture in vitro MSCs from bone marrow of the adult rats by density gradient centrifugation combined with adherence culture, and observe the growing characteristics and the possibility of mass multiplication.DESIGN: A completely randomized grouping design/repetitive measuring experiment.SETTING: Pathological laboratory, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: Four six-week-old SD rats about 250 g, grade Ⅱ of cleaning, were provided by the Animal Center of Sun Yat-sen University [certificate number: SCSK(Yue)2004/0011], about 250 g each rat and there was no limit to the sex. The main reagents and instruments included low sugar Dulbecco modified Eagle culture medium (DMEM/F12, American Gibco Corporation), trypsin (fetal bovine serum (FBS, Hangzhou Sijiqing Bio-Engineering Material Research Institute), American Gibco Corporation), disodium edetate, lymphocyte separating medium, fibronectin, CD44, CD34, CD31 monoclonal antibodies, two-step-method kit for immunohistochemistry (Beijing Zhong shan Biotechnology Corporation).METHODS: This experiment was conducted at the Key laboratory of Ophthalmology (Sun Yat-sen University), Ministry of ethanol (750 g/L) for 10 minutes. Under aseptic condition, the medullaris cavitas was exposed, the syringe containing application m edium was directly punctured into the femoral cavity, the cells in the medullaris cavitas were washed out with the culture medium containing heparin and taken as the cell suspension. The bone marrow-derived MSCs were separated and purified by density gradient centrifugation combined with adherence culture, and the growing conditions of the wells. When the cells had generally connected with each other, they were fixed with methanol or dimethy ketone in situ for 10 minutes, and then hematoxylin eosin (HE) staining or immunohistochemical staining. Antibodies against fiinoculated to 96-well culture plate by a cell density of 4.25×107/L, with 200 μL every well; then put the culture plate into culture box. Then from the next day to the sixth day, 5 g/L MTT solution was added into two rows (20 μL every well) every day, continuously cultured for 4 hours, then the supernatant was removed, and 200 μL DMSO was added to each well, agitated for 5 minutes, then detected the absorbance (A) values at 570 nm wave length, and a growth curve was drawn.branes were clear and the cell bodies were lucent. Being cultured for 2 days, there appeared adherent cells. 3 days later,most of the adherent cells extended and appeared to be in polygon, star or long-shuttle shapes. 4 days later, the cells showed to be in division growth stage; and about 12 days later, the cell clones were connected to each other, appearing curve of subcultured MSCs was in S shape. Cells began growing fast on the 2nd day after being passaged, and they entered the growth period for 3-4 days followed by saturation period, and then cells stopped growing.CONCLUSION: It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of density gradient centrifugation and adherence. MSCs can greatly proliferate in vitro and offer seed cells for the application of tissue engineering technique to treat eye diseases.