Objective To analyze the curative effects of total knee arthroplasty(TKA)for severe genu varus combined with flexion deformity.Methods The clinical data of 25 patients(36 knees),who had undergone TKA for severe genu varus combined with flexion deformity from January 2005 to October 2010,were retrospectively analyzed.There were 7 males and 18 females,aged from 55 to 80 years(average 70.5 years).The primary diseases were osteoarthritis in 22 cases and rheumatoid arthritis in 3 cases.HSS knee score was used before and after the operation to assess the efficacy of the TKA.Results All of the patients were followed up for 4 to 9 years(average 6 years).The degree of flexion deformity was reduced from (21 ±63)°to(1.1 ±2.3)°;the degree of the varus decreased from(210 ±4.8)°to(175.6 ±2.1)°;range of motion of the knee joint increased from(70.5 ±20.5)°to(115.1 ±5.3)°;knee score augmen-ted from(33.2 ±10.5)to(90.7 ±8.5);function score added from(35.5 ±14.2)to(85.6 ±10.5);there were 21 cases(8 knees)rated as excellent,2(3 knees)as good and 2(3 knees)as fair,and the excel-lent and good rate was 86%.Most of the patients had normal force line of the knee joint,but 2 patients re-mained varus deformity of 5°~10°.Conclusion Satisfying outcome can be achieved by TKA in treating severe genu varus combined with flexion deformity of the knee.

ObjectiveTo explore the clinical effect of levofloxacin used on patients with HAP. Methods134 cases of patients with HAP as studied object were randomly divided into observation group and control group,observation group were given therapy of levofloxacin,and control group were given therapy of azithromycin. ResultsThe efficiency cases,effective cases,vain cases in observation group were 51,13 and 3,the efficiency rate and total effective rate were 75.0％ and 93.3％, the data in control group were 38,17,12,51.7 ％ and 81.7 ％. The efficiency rate ( x2 =4. 818, P ＜ 0.05 )and total effective rate( x2 =4. 804 ,P ＜ 0.05 ) of observation group was significantly better than that of control group. ConclusionLevofloxacin used on patients with HAP, could acquire good clinical effect,and the side-reactions were light.

Objective To study the effect of radiofrequency ablation (RFA) with sublethal temperature on the production of liver cancer stem cells (LCSCs) and on the expression of LCSCs-related transcriptional factors.Methods Mouse hepl-6 hepatoma cell line and clinical samples of patients with hepatocellular carcinoma (HCC) were used to test the expressions of LCSCs-related markers and transcriptional factors.Results Different temperatures were used to stimulate Hep1-6 cells,and it was proved that the temperature of 45℃ was a sublethal temperature that could not induce cell death.Flow cytometry testing showed that treatment with 45℃ could obviously increase CD13+,CD44+,CD90 and CD133+ Hep1-6 cells,suggesting that treatment with 45℃ could increase the production of above mentioned types of LCSCs in hep1-6 cells.Real-time quantitative polymerase chain reaction (RT-qPCR) assay indicated that the temperature of 45℃could cause significant increase in CD13,CD90 and CD133 mRNA.In all 5 HCC patients,CD13 mRNA in the recurrent HCC lesions was remarkably increased,CD133 mRNA was increased in 4 patients with recurrent HCC,and CD90 mRNA was increased in only one patient with recurrent HCC.Flow cytometry testing revealed that CD13+ LCSCs were strikingly increased in 4 recurrent HCC patients,while CD133+LCSC was increased in only one patient,suggesting that more close correlation existed between the increase of CD13+ LCSCs and the temperature of 45℃.RT-qPCR assay showed that in 4 recurrent HCC patients with increased CD13+ LCSC,the Sox2 and Stat2 among 13 LCSCs-related transcriptional factors were obviously increased.Flow cytometry testing showed that 45℃ treatment also increased the expression of Sox2 and Stat1 mRNA in Hep1-6 cells.Finally,Sox2 and Stat1 could be knockdown by siRNAs,indicating that both Sox2 and Stat1 transcriptional factors were involved in 45℃-induced production of CD13+ LCSCs in Hep1-6 cells.Conclusion In RFA therapy,the use of sublethal temperature of 45℃ can increase CD13+LCSCs,which is related to the promotion of Sox2 and Stat1 expression.The results of this study can be used for reference in the research of liver cancer recurrence.

Objective To investigate the clinicopathological significances of LDHA/mutant p53 co-expres-sion in gliomas. Methods According to the 2016 WHO CNS,archived 68 gliomas were collected and analyzed retrospectively. The co-expression of LDHA/mutant p53 was detected by immunohistochemical staining. Results High expression of LDHA alone was always found in high grade gliomas(48.5%). Mutant p53 high expression was usually observed in glioblastomas (26.5%). There was a close relationship between co-expression of LDHA/mutant p53 in glioblastoma(27.9%,P = 0.005),or gliomas with high histological grading(27.9%,P = 0.002). Conclusions Co-expression of LDHA/mutant p53 in tumor cells might be a specific immunohistochemical pheno-type of gliomas,and may help for distinguishing glioblastoma and other high grade gliomas from low grade gliomas.

Objective To construct the bait expression plasmid pGBKT7-GR of glucocorticoid receptor(GR)binding domain.Methods The fragments of GR binding domain was amplified by RT-PCR,and then was cloned into pMD18-T.After being verified by sequencing,it was subcloned into the bait expression vector pGBKT7.Then the bait vector pGBKT7-GR was transformed into AH109 yeast cells and the expression of the bait protein was analyzed by Western blot.Toxicity and self-activation of the bait protein were detected.Results GR binding domain was amplified and cloned into pMD18-T and pGBKT7 successfully.The bait vector was transformed into AH109 yeast cells successfully,without toxicity or self-activation.The expression of the bait protein was confirmed by Western blot.Conclusion The successful construction of bait expression vector of glucocorticoid receptor binding domain lays the foundation for constructing small molecule ligand yeast three-hybrid system.

Aim To study the mechanism of cucurb-itacin E ( CuE )-induced autophagy in HeLa cells. Methods Improved MTT assay was adopted to meas-ure the effect of CuE on cell proliferation. Western blot was used to determine the phosphorylation levels of downstream signaling proteins of mTORC1 and the ex-pression of autophagy associated proteins. ResultsCuE inhibited the proliferation of HeLa cells in a dose-dependent manner, and the 24-h IC50 of CuE was 4. 01μmol· L-1 . CuE significantly inhibited the phospho-rylation of p70 S6 K in a time-and dose-dependent man-ner as evidenced by decreased phosphorylation levels of
the mTORC1 substrate. Meanwhile, the expression of LC3-II, a marker for autophagosome formation, was elevated by CuE treatment, and was further increased in the presence of chloroquine. Furthermore, CuE re-duced the levels of p62/SQSTM1 . These results indi-cated that CuE induced autophagy in HeLa cells. The decreased levels of phosphorylated ULK1 S757 were posi-tively correlated with autophagy induction in HeLa cells. Conclusion CuE is likely to induce autophagy through inhibiting mTORC1 activity.

Objective To detect the incidence of inherited metabolic diseases(IMD)and disorders of metabo-lism in 4 710 high - risk infants,as well as providing basis of clinical diagnosis and treatment by using urease pretreat-ment - gas chromatography - mass spectrometry(UP - GC - MS). Methods Samples were collected from high - risk infants with IMD,after removing urea,putting in internal standard,removing protein,vacuum drying and bis (trimethyl - silyl)trifluoroacetamide / trimethyl - chlorosilane derivativing,UP - GC - MS was used to analyze compo-sitions such as organic acids,amino acids,carbohydrates,pyridoxines,purines and pyrimidines,then metabolic analysis was proceeded to refer to the normal detection value of the healthy children,finally a metabolic diagnosis was made ba-sing on the clinical data such as the high - risk clinical manifestations and general biochemical tests and other special examinations. Results In the 4 710 cases,there were 98 cases of IMD(2. 1% ),326 cases of suspected IMD(6. 9% ), 2 610 cases of metabolic disorders(55. 4% ). There were 98 cases of IMD,including 57 cases of methylmalonic aciduria,12 cases of propionic acidemia,7 cases of glutaric aciduria,5 cases of hyperphenylalaninemia,maple syrup u-rine disease and multiple carboxylase defects each,4 cases of isovaleric acidemia and 3 cases of 4 - hydroxy butyric acid urine disease. Conclusions UP - GC - MS is a effective way to diagnose IMD and metabolic disorders of infants. Common IMD in Guangdong Province include methylmalonic aciduria,propionic academia,glutaric aciduria,hyperphe-nylalaninemia,maple syrup urine disease and multiple carboxylase defects. The results of the tests can provide effective guidance for diagnosis and treatment of suspected infants.

Objective To evaluate the application of echocardiography in guiding percutaneous balloon pulmonary valvuloplasty in children and to summarize the key echocardiographic planes used in the procedure Methods From February 2013 to September 201 5 38 isolated congenital pulmonary valve stenosis patients were recruited Case inclusion criteria age ≥3 years old purely congenital pulmonary valve stenosis and pulmonary transvalvular pressure gradient ≥40 mmHg Echocardiography was used to assess the severity of pulmonary valve disease and to measure pulmonary transvalvular pressure gradient before procedure Intraoperative transthoracic or transesophageal echocardiography was used to monitor the whole process of percutaneous balloon pulmonary valvuloplasty and to evaluate immediate postoperative efficacy of the procedure All patients were followed up by echocardiography after a month post-discharge Results Thrity eight cases were successfully treated by echocardiography-guided percutaneous balloon pulmonary valvuloplasty The average age of children was 7 1 ±2 5 years mean body weight was 25 3 ±7 1 kg Before the procedure pulmonary transvalvular pressure gradient was 65 9 ± 8 9 mmHg pulmonary annular diameter was 14 6±1 1 mm Immediate postoperative pulmonary transvalvular pressure gradient was 1 5 5 ± 3 4 mmHg All children survived and had no significant complications After a month pulmonary transvalvular pressure was 16 1 ± 3 3 mmHg Conclusions Echocardiography plays an important role in percutaneous balloon pulmonary valvuloplasty for children with congenital pulmonary valve stenosis As a non-x ray guided way it has advantages in preoperative screening of patients intraoperative real-time monitoring and postoperative assessment of efficacy The key sections of echocardiography for intraoperative monitoring are four-chamber and aortic short axis view.

Objective To prospectively evaluate the value of US,MSCT,EUS and MRI in the quantitative evaluation of the extent of pancreaticobiliary duct obstruction in pancreatic cancer.Methods Consecutive 68 patients with pancreatic carcinoma underwent US,MSCT,EUS and MRI before surgery.The diameter of extrahepatic bile duct and pancreatic duet were measured,and correlation analysis was performed with surgical specimens.Results Diameters of extrahepatic bile duct scaled by US.MSCT,EUS and MRI were(16.60±6.33)mm,(18.90±6.74)mm,(18.80±5.88)nun and(17.26±4.83)mm,and diameter measured from surgical specimens was(18.39±6.05)mm;the correlation among the four imaging examinations and the surgical evaluation were r=0.3839,P=0.1055;r=0.7113,P=0.0011; r=0.3759,P=0.0465;r=0.3376,P=0.2872,respectively. Kappa Values were 0.6285,0.7115,0.6661 and 0.7490,respectively.The diameter of pancreatic duct was(15.90±3.41)mm,(6.83 4-3.70)mm,(6.77±3.22)mm and(5.58±2.65)mm,and diameter measured from surgical specimens was(5.97±2.60)mm,the correlation among the four imaging examinations and the surgical evaluation were r=0.3584,P=0.2895;r=0.6148,P＜0.0001; r=0.7373,P＜0.0001;r=1.0746,P＜0.0001.Kappa values were 4.159,9.094,9.001 and 4.050.All of these parameters were in coherence with surgical findings.Condusions US could be used as the initial method in the assessment of extrahepatic and pancreatic duct obstruction.MRI and MSCT,combined with EUS if necessary,could be used to quantitatively evaluate the extent of pancreaticobiliary obstruction.