Objective To investigate the relationships between the expression of VEGF,its receptor KDR/flk-1,cNOS mRNA and angiogenesis,cell proliferation,metastasis in human hepatocellular carcinoma(HCC). Methods Immunohistochemical analysis using antibodies against VEGF and its kinase insert domain receptor (KDR) was carried out.cNOS mRNA expression in HCC,liver cirrhosis and normal liver tissue was observed by in situ hybridization.CD34 immunostaining was used to measure the microvascular density(MVD)and proliferative index was evaluated by Ki-67 immunostaining. Results The expressions of VEGF and KDR of HCC were significantly related to MVD,proliferation and metastasis of HCC(P

Objective To investigate the association of angiotensinogen(AGT)gene A-6G、T174M and G-217A polymorphisms with the risk of essential hypertension(EH)in the elderly of Han nationality.Methods Genotypes of AGT gene A-6G,T174M and G-217A polymorphisms in 177 aged EH patients and 86 sex and age-matched controls were analyzed with gene chip technology.Results The A-6G and T174M polymorphisms of AGT gene were significantly associated with EH.The numbers of the three genotypes of A-6G were 113,58 and 6 in the patient group and 70,15 and 1 in the control group(P= 0.014)and those of T174M were 94,77 and 6,60,25 and 1(P=0.031),respectively.G-217A polymorphism was not related to EH.Individuals carrying A-6G AA and T174M CC genotypes showed 57% and 56% lower risk of EH(OR=0.43;95%CI=0.23-0.82 and OR=0.44;95%CI=0.25-0.79, respectively).Conclusions The A-6G AA and the T174M CC genotype may be related with decreased risk of EH and G-217A polymorphism may have little role in the etiology of EH in Han nationality.

OBJECTIVETo investigate clinical outcomes of tendon allograft reconstruction with arthroscopy minimally invasive technique at stage I for the treatment of knee dislocation with multiple ligaments injury.

METHODSForty-eight patients with knee dislocation were reconstructed anterior and posterior ligament under arthroscopy at stage I from January 2008 to January 2012, and repaired ligaments injury of knee joint by minimally invasive technique. There were 38 males and 10 females aged from 20 to 59 years old with an average of 35.6 years old; 22 cases on the left side and 26 cases on the right side; the time from injury to operation ranged from 2 d to 2 weeks. Two cases combined with anterior cruciate ligament (ACL), posterior cruciate ligament (PCL), medial collateral ligament (MCL) and posterolateral complex injuries, 36 cases combined with ACL, PCL, and MCL injuries, 10 cases combined with ACL, PCL and PLC injuries; 4 cases combined with peroneal nerve injury. Lysholm scoring were used to compared the cases before operation and final following-up to evaluate knee function.

RESULTSAll patients were followed up from 12 to 30 months with an average of (18.2 ± 6.3) months. Activity and stability of joint were obviously improved. Lysholm score were improved from 40.3 ± 4.1 before operation to 87.0 ± 6.4 at final following-up.

CONCLUSIONReconstruction with arthroscopy minimally invasive technique at stage I for the treatment of knee dislocation with multiple ligaments injury could recover stability of joint better,reserve joint function. Preoperative training and postoperative individualized rehabilitation treatment is the key point of recover knee joint function.

Adult ; Anterior Cruciate Ligament Injuries ; Arthroscopy ; Female ; Humans ; Knee Dislocation ; rehabilitation ; surgery ; Male ; Middle Aged ; Multiple Trauma ; surgery ; Posterior Cruciate Ligament ; injuries ; Reconstructive Surgical Procedures ; methods

Objective To explore the pathological changes by CT scan on localized thermochemotherapy dur- ing and after the operation of human gliomas.Methods Retrospective analysis was given to the CT scan of 37 pa- tients receiving thermochemotherapy during and after the operation,and the relation of the tumorous cells and mi- crovessels and CT density by EM were analyzed.Changes of tumorous cells and microvessels after localized ther- mochemotherapy on C_6 gliomas in rat were analyzed.Results When the tumor was low dense on CT pattern,less cellular number with increasing the amount of fluid between the cells was demonstrated pathologically.On EM,a lower cellular electron density was observed.The amount of fluid in cytoplasm was increased,the cytoplasm was porous,swelling denaturation was chiefly seen in organelle.If the tumor had mixed density on CT,cellular number was more,the amount of fluid was less.On EM,cellular electron density increased correspondingly,the fluid in cyto- plasm decreased,organdie was aggregated.After thermochemotherapy,the tumor reduced,liquefied,and vanished by CT scan.It could be observed that the tumorous cell become smaller,concentrated and cataclased,finally formed apoprotic bodies and separated from the cell in C_6 gliomas in EM.The tumorous vessels was less,smaller and thinker. Some vessels only could see the base membrane and no endothelioid cells.Conclusion The remaining tumors is van- ished by CT scan.The mechanisms of tumors disappearance proposes to explain that thermochemotherapy can dam- age C_6 glioma cells and microvessels,decrease microvessels density and induce tumor ceils apoptosis.That inhibits tu- morous angiogenesis and proliferation.

OBJECTIVETo analyze the Chinese medicine syndrome types of patients undergoing maintenance peritoneal dialysis to provide some clinical reference for the treatment based on syndromes.

METHODSAccording to the criterion made by the Nephropathy Branch of China Association of Chinese Medicine in 2006, the syndrome type of 156 patients were differentiated, and the related laboratory parameters, including serum albumin (ALB), C-reactive protein (CRP), hemoglobin (HB), total urea clearance rate (KT/Vt), residual kidney urea clearance (KT/Vr), blood flow mediated vascular endothelial dilatation (FMD) and volume overload (OH) were measured.

RESULTSSyndrome type presented in patients was different. Along with the progress of dialysis, it changed in the root syndromes from qi-deficiency to yang-deficiency and further to both yin-yang deficiency, while in the superficial syndromes it turned from turbid-damp to blood-stasis. ALB in patients with Pi-shen yang-deficiency type and both yin-yang deficiency type was significantly lower than that in patients with Pi-Shen qi-deficiency type and both qi-yin deficiency type (P < 0.05); KT/Vt in both yin-yang deficiency type was the lowest, significantly lower than that in Gan-Shen yin-deficiency type and both qi-yin deficiency (P < 0.05); OH in Pi-Shen yang-deficiency type and both yin-yang deficiency type was significantly higher than that in other types (P < 0.01). Comparison of patients' age showed that group of patients without superficial syndrome was the youngest and the group of patients with damp-heat syndrome type was the oldest (P < 0.01). CRP in damp-heat type was significantly higher than that in other types (P < 0.05); FMD was lower in blood-stasis type than in turbid-damp type and toxic heat type (P < 0.01); and OH was significantly higher in turbid-damp type than in other types (P < 0.01).

CONCLUSIONSome rules of syndrome type distribution could be seen in patients undergoing peritoneal dialysis, which is related with some laboratory parameters to a certain extent, so may provide a few references for clinical treatment based on syndrome type.

Adult ; Aged ; Aged, 80 and over ; C-Reactive Protein ; metabolism ; Female ; Humans ; Inflammation ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Peritoneal Dialysis, Continuous Ambulatory ; Serum Albumin ; metabolism

BACKGROUND: Angiotensinogen (AGT) gene is the firstly discovered candidate gene for essential hypertension, both the T174M and M235T polymorphisms locate at the second exons of AGT gene, and there is existence of linkage disequilibrium. The polymorphism at A-6G and G-217A sites in promotor region plays an important role in regulating the gene expression, and the products of keep close correlation with the level of blood pressure. OBJECTIVE: To investigate the association between the polymorphism of AGT gene at A-6G, T174M and G-217A sites and the risk for the attack of essential hypertension in Chinese Han population, DESIGN: A cluster sampling and case-control analysis. SETTINGS: Department of Geriatrics and Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University; Southern Research Center of National Human genome; Department of Cardiology, Dongtai People's Hospital of Jiangsu Province. PARTICIPANTS: The experiment was carried out in the countryside of Dongtai county, Yancheng city, Jiangsu province. All the subjects were selected from the countryside of Dongtai county, Yancheng city, Jiangsu province. Totally 177 patients with essential hypertension who had never accepted any drug treatment, were taken as the essential hypertension group, and hypertension was diagnosed according to the diagnostic standard of hypertension set by WHO/ISH in 1999 (systolic blood pressure ≥ 140 mm Hg and/or diastolic blood pressure ≥ 90 mm Hg); Another 86 normal person were taken as the normal control group. ② Inclusive criteria: The enrolled subjects should be Han nationality; long-term local residents but not from other places; able to answer questions clearly; diagnosed by disease history, clinical symptoms, physical signs and assistant examinations; have complete data of investigation of uniform questionnaires by face-to-face interview (including demographic information, profession history, family history and life styles of smoking, drinking, drinking tea, etc.). ③ Exclusive criteria: The patients with secondary hypertension in the essential hypertension group, subjects having family hisory of hypertension in the normal control group, and those with chronic diseases of liver and kidney, and diabetes mellitus in both groups were excluded. METHODS: Peripheral venous blood samples (3 mL) were collected, and DNA was extracted from human peripheral blood with FlexiGene DNA Kit (250). The Primer3 software was applied to design primers, and the polymorphism sites in the primer sequence were excluded. After multiplex polymerase chain reaction (PCR), 3 μL products were selected to detected the amplified results by agarose gel electrophoresis. The successfully amplified PCR products were purified with the QIAquick PCR Purification Kit, and the purified products were fragmentized with Dnase Ⅰ . The fragmentized products of enzyme digestion were labeled with fluorescein by deoxynucleotide terminal transferase. Two allele specific probes and one mismatched probe were designed respectively for each single nucleotide polymorphism. The chips were prepared with the OmniGridTM 100 TLC samler, each probe was repeated for three times to form three matrix. The hyridization solution was degenerated at 95 ℃ for 10 minutes, and then immediately cut on ice. 10 μL hybridization solution was added onto the chip matrix, hybridized at 50 ℃ for 2 hours, then washed and dried. The chips were scanned with the GenePix 4000B laser confocal scanner (Figure 2),and the intensity of the fluorescent signal for each probe was extracted with GenePix Pro, and the allele score of each single nucleotide polymorphism was calculated to judge the genotype. MAIN OUTCOME MEASURES: ① Comparison of the frequencies of genotype distribution at each polymorphism site of AGT gene in both groups; ② Correlation analysis of the polymorphism of AGT gene at A-6G and T-174M sites with the risk for the attack of essential hypertension; ③ Effects of the polymorphism of AGT gene at A-6G, T-174M and G-217A sites on blood pressure.RESULTS: According to the intention-to-treat analysis,all the 263 subjects were involved in the analysis of results. ① At the A-6G site of AGT gene, the frequencies of AA, AG and GG genotypes (P=0.014) and A and G alleles (P=0.004, OR=0.44) had significant differences between the essential hypertension group and normal control group; At the T174M site, the frequencies of CC, CT and TT genotypes (P=0.031) and A and G alleles (P=0.014, OR=0.55) were significantly different; At the G-217A site, no obvious differences were found in the GG, AG and AA genotypes (P=0.722) and G and A alleles (P=0.403, OR=0.80). ② The risk of essential hypertension in the individuals carrying AA genotype of A-6G polymorphism and CC genotype of T174M polymorphism was reduced by 57% (95%CI= 0.23-0.82, P= 0.010) and 56% (95%CI= 0.25-0.79, P= 0.006) respectively. ③ There were no significant differences in the systolic blood pressure, diastolic blood pressure and mean arterial pressure among different genotypes at the A-6G, T174M sites and G-217A sites (F=0.100- 2.911, P ＞ 0.05). CONCLUSION: The AA genope at A-6G and the CC genotype at T174M site of AGT gene may reduce the risk for the attack of essential hypertension in Chinese Hun population, and no significant correlation was found between the genotype of G-217A polymorphism and the attack of essential hypertension.

Objective To analyze the related factors associated with the effect of Roux-en-Y gastric bypass in Goto-Kakizaki rats.Methods In GK rats undergoing Roux-en-Y gastric bypass,the weight,food intake,and the fasting blood glucose were retrospectively analyzed before surgery and 1,3,6,12,24 weeks after surgery,and the glycosylated haemoglobin concentrations were retrospectively analyzed before surgery and 4,12,24 weeks after surgery.According to preoperative glycosylated haemoglobin concentrations,GK rats were divided into 3 groups(group A,HbAlc 6.5％-7.9％; group B,HbAlc 8.0％-9.9％ ; group C,HbAlc ＞ 10％).Results The fasting blood glucose and the glycosylated haemoglobin concentrations significantly decreased 1 week after surgery(P ＜ 0.01),on 24 weeks after operation,the fasting blood glucose levels decreased from(12.1 ± 3.0)mmol/L to(7.6 ± 1.3)mmol/L,and the glycosylated haemoglobin concentrations declined from(9.2％ ± 1.8％)to(6.3％ ± 0.8％).Preoperative fasting blood glucose was(11.1 ± 2.2)mmol/L and(15.8 ± 2.3)mmol/L respectively,and the preoperative glycosylated haemoglobin concentration was(8.6％ ± 1.4％)and(11.5％ ± 1.4％)respectively(P ＜ 0.01).Non-conditional Logistic regression analysis identified preoperative glycosylated haemoglobin concentrations as an independent predictor for the cure rate(P ＜ 0.01); Differences were significant between group A and group C,and group B and group C on the efficiency of the surgery(P ＜0.01).Conclusions Significant correlation established between glycosylated haemoglobin concentrations and the efficiency of the surgery,especially for those rats with preoperative HbAlc ＜ 10％.

Objective To investigate the blood and chest fluid level of Th17 cell and IL-17 in patients with tuberculous pleural effusion and its pathological role.Methods Flow cytometry and enzyme linked immunosorbent assay (ELISA) were used to measure the blood and chest fluid level of Th17 cell and IL-17 from 30 patients with tuberculous pleural effusion,20 patients without tuberculous pleural effusion,and 20 healthy persons.Results The blood level of Th17 cell and IL-17 wwere higher in tuberculous pleural effusion than in the other two groups(P ＜0.05).The chest fluid level of Th17 cell and IL-17 in patients with tuberculous pleural effusion were significantly higher than those in patients without tuberculous pleural effusion (P ＜ 0.05 ).The chest fluid level of Th17 cell and IL-17 in patients with and without tuberculous pleural effusion were significantly higher than that of the blood serum level.After treatment for 1,3,7 and 14 days,tbe blood serum and chest fluid level of Th17 cell and IL-17 were obviously lower.( P ＜ 0.01 ).After treatment for 1 day,the blood level of Th17 cell was obviously lower than before treatment( P ＜ 0.01 ).After treatment for 3 days,the chest fluid level of Th17 cell was obviously lower than before treatment( P ＜0.01 ).After treatment for 3 days,the blood serum level of IL-17 was obviously lower than before treatment (P ＜ 0.01 ).After treatment for 7 days,the chest fluid level of IL-17 was obviously lower than before treatment ( P ＜0.01 ).Conclusion Th17 cell and IL-17 might play an important role in the pathogenesis of tuberculous pleural effusion and they were correlated with disease progression and the therapeutic effect.

Objective Multiple carboxylase deficiency(MCD) is an autosomal recessively inherited defect of organic acid metabolism.The underlying mechanism is biotinidase(BT) or holocarboxylase synthetase(HLCS) deficiency.The authors reported 15 cases of MCD(clinical characteristics,diagnosis and treatment) and outcomes of long-term follow-up.Methods The clinical data of 15 patients with MCD admitted to Guangzhou Women and Children's Medical Center between Aug.2001 and Feb.2013 were analyzed.The diagnosis was confirmed by urinary organic acid analysis with gas chromatography/mass spectrometry (GC/MS),blood enzymatic determination and gene analysis.The patients consisted of 12 male and 3 female.The onset age ranged from newborn infants to 16 months.Results 1.Remarkable elevations of urinary lactate,3-oxy-butyric acid,3-OH-isovalerate,methylcitrate,3-methylcrontonylglycine,3-OH-propionate were detected in all of 15 cases.Fourteen cases with HLCS deficiency and 1 case with BT deficiency were confirmed by gene analysis.2.Most of patients with HLCS deficiency presented in the neonatal period or early infancy.The main clinical manifestations were skin rash (14 cases),tachypnea (9 cases),developmental retardation (8 cases),vomiting(5 cases),poor feeding (3 cases),developmental regradation (1 case),convulsion (1 case).Laboratory evaluation showed persistent metabolic acidosis and varied degree of ketosis,lactic acidosis,hyperuricacidemia,ammoniemia and hypoglycemia.Biotin was given to 13 patients in 10 mg/d and their metabolic disorders were corrected within 48 h,except one who gave up treatment and died.Treatment with Biotin in 5 mg/d provided clinical stability and normal neurodevelopmental outcome on follow-up for 3-11 (6.47 ± 2.70) years.3.One patient with BT deficiency presented with severe neurological symptoms(impaired consciousness,limbs trembling,tachypnea with irregular respiratory rhythm) at 16 months old.Demyelination of corpus callosum was evident on magnetic resonance imaging.Biotin treatment was given to him on the second of onset,and 1-year follow-up indicated significant improvement of his clinical symptoms,but the right limb weakness did not completely recover.Conclusions The main clinical features of HLCS deficiency include frequent skin rash,tachypnea,and metabolic disorders.BT deficiency may produce variable neurologic manifestations including demyelination of corpus callosum.Urinary organic acid analysis with GC/MS is critical to the early diagnosis of MCD.Prompt biotin treatment is recommended to correct metabolic derangements and continued therapy is essential to the improvement of long-term prognosis.Delayed commencement of therapy in BT deficiency can result in irreversible neurological damage.

Objective To investigate the expression and distribution of plasmacytoid dendritic cells(pDC) in peripheral blood and renal tissues in children with Henoch-SchSnlein purpura(HSP),and explore the role of pDCs in the pathogenesis of Henoch-Schtnlein purpura nephritis(HSPN).Methods Among the 40 children with HSP,28 cases were in the active phase(renal biopsy performed in 8 cases of them) and the other 12 in remission phase.Peripheral blood mononuclear cells were isolated,and the expression of pDC was detected by flow cytometry.The normal control group was established (n =15).Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression(indicated as 2-△Ct value) of CXC motif chemokine 10 (CXCL10),CC chemokine ligand 5 (CCL5),chemokine CXC subfamily receptor 3 (CXCR3),CC chemokine receptor 5 (CCR5) in children with HSP and those in the controls.Immunohistochemistry labeling technique was used to detect the distribution of pDC in renal tissues from renal biopsy,and the normal controls were established (n =3).Results The expression percentage of pDC in peripheral blood in active phase was 0.051 ± 0.039,significantly lower than those in remission phase (0.181 ± 0.082) and the normal controls (0.166 ± 0.079) (P ＜ 0.000 1).Chemokines genes CXCL10 and CCL5 were overexpressed in peripheral blood ceils of acute phase HSP children,but chemokine receptors CXCR3,CCR5 were lowly expressed compared with normal controls.There was almost no expression of pDC in the normal control renal tissues,while pDC was infiltrated in glomeruli of HSPN children.Conclusions The number of pDC and chemokines' expression in peripheral blood is abnormal,and the pathogenesis of nephritis may be involved with the pDC in peripheral blood to migrate to the renal tissues.