Objective: To determine if fusion genes of HSV-tk gene and cytokine gene have synergy on the cell killing of the Hep-2 human laryngeal carcinoma cell line in vitro. Methods: Different fusion genes expressing vectors PL(TI) SN, PL(TT)SN and PL(TK)SN were generated by recombinant DNA technology. Hep-2 was infected by the recombinant retrovirus. The positive clones were obtained after G418 selection and were termed Hep/TI, Hep/TT and Hep/TK respectively. The integration and expression of fusion genes in Hep-2 cells were identified by RT-PCR and Southern blot. The growth state and GCV killing effect of fusion genes modified cells were used to investigate the expression of fusion genes and antitumour effect on Hep-2 cells. Results: RT-PCR and Southern blot analysis confirmed the integration and expression of fusion genes in Hep-2 cells. There was no significant difference in cell proliferation between the Hep/TI and Hep/ TK.but the growth of Hep/TT was restrained. After the treatment of GCV the Hep/TI, Hep/TT and Hep/TK all showed high sensitivity to GCV. The killing effect of GCV on Hep/TT was the most siginificant and bystander effects were observed siginificantly in vitro. Conclusion: The fusion genes of HSV-tk and cytokine gene have synergistic effects on killing Hep-2 cell after treatment of GCV in vitro,which might have therapeutic potentials for laryngocarcinoma.

Cardiomyopathies ; etiology ; Humans ; Polymyositis ; complications

Objective: To construct the eukaryotic expression vector pSurp-EGFP regulated by the survivin gene promoter and to detect the specific expression of the promoter in human laryngeal cancer Hep-2 cells by green fluorescent protein assay.Methods: Thesurvivin gene promoter was generated by polymerase chain reaction(PCR) and the CMV promoter of the pShuttle vector replaced by the survivin gene promoter to generate the plasmid pSurp.The three plasmids pShuttle,pSurp and pEGFP-C1 were respectively double-enzyme digested so as to produce the plasmids pCMV-EGFP and pSurp-EGFP carrying the CMV or survivin promoter.The purified pCMV-EGFP and pSurp-EGFP were transfected into Hep-2 cell and vascular endothelial cell ECV304 using liposome transfection reagent and the expressions of EGFP detected by the fluorescent microscope.Results: Thesurvivin gene promoter was successfully cloned by PCR,and thesurvivin gene promoter-regulated pSurp-EGFP was constructed.Green fluorescence was observed in Hep-2 cells but not in ECV304. Conclusion: The high specific activity of the survivin gene promoter in Hep-2 cells that we successfully constructed attributes to the studies of tumor specific gene therapy.

AIM: To investigate lipoprotein lipase gene HindⅢ polymorphism and its relationship with serum lipids and apolipoprotein, serum HDL subclasses in patients with hyperlipoidemia. METHODS: Lipoprotein lipase gene HindⅢ polymorphism was assayed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The subclasses of serum HDL in 152 hyperlipoidemia patients and 128 healthy subjects were determined by two-dimensional gel electrophoresis conjunction with immunodetection method. RESULTS: H+H+ genotype and allele H+ in hyperlipoidemia and control groups were both the highest. In hyperlipidemia group, H+H+ genotypes tended to be higher than that in control group, while H+H- and H-H- genotypes were significantly lower (P

Objective To explore the microanatomy of the horizontal segment of the facial nerve (FN)in temporal bone and its adjacent structures,in order to provide microanatomical datag for the clinical works.Methods In 20 adult wet skull specimens(40 temporal bones),horizontal segment of FN and their adjacent structures in temporal bones were observed according to operational layers by simulating transmagtoid approach.Results The lenglh of the horizontal segment was[(8.85±1.01)(7.10～11.25)]mm;The diameter of the horizontal segment wag[(1.88±0.65(1.55～1.90)]mm;The angle opening towards anterior direction between horizontal segment and vertical segment wag[(115.5±6.89)(109.5～128.6)]..The ansh towards posterior direction between horizontal segment of FN and tympanic tegmen wag [(28.5±3.66)(25.8～31.5)°;The vertical distance from the cochlear form process to the horizontal segment of the FN wag[(1.89±0.58)(0.90～3.05)]mm;The vertical distance from the head of the stapes to the horizontal segment of the FN wag[(2.30±0.85)(1.97～3.11)]mm;The distance between the summit of pyramidal segment of FN and the apex of shor limb of incus wag 2.55±0.21(2.10-2.90)mm;The distance between the summit of pyramidal segment of FN and the eminence of the lateral semicircuiar canal wag[(2.86±0.31)(2.23～3.56)]mm;No branching or dislocated nerve of the horizontal segment of the FN wag found in all 40 cases.Five percent(2/40)distal part of the horizontal segment of FN tracks forward lateral to the eminence of the literal semicircular canal.Conclusion Geniculate ganglion,cochlearform process,stapes,tympanic tegmen,lateral semicircular canal,incus are important landmarks by which horizontal segment of FN might be located in manipulation of mastoid;The relations of horizontal segment of FN and its adjacent structures are complicated,compact.It will be good that the manipuliter absorb a great deal of anatomical knowledge about landmarks that can be used in locating horizontal segment of FN.

Objective To explore the clinical features and treatment of thyroid carcinoma in children. Method The clinical data of 19 children under 14 years old with thyroid carcinoma diagnosed and treated from January 2003 to January 2014 were retrospectively analyzed. Results In 19 cases (12 males and 7 females), there were 18 cases of papillocarcinoma and one case pf follicular carcinoma. Unilateral lobectomy plus isthmectomy was performed in 6 cases, subtotal thyroidectomy in 4 cases and total thyroidectomy in 9 cases. Unilateral cervical lymph node dissection was performed in 5 cases and bilateral in 11 cases. After the operation, multiple lesions were confirmed by pathology in 9 cases, thyroid capsular invasion in 14 cases, lymphatic metastasis in 15 cases and distant metastasis in 5 cases. All the patients were treated with TSH, and 10 cases were treated with 131I after operation. The median follow-up time was 63 months. There was no death in all cases, while local residual tumor recurrence was found in 2 cases and cervical lymph node metastasis in 2 cases and distant metastasis in one case. Conclusion Thyroid carcinoma in children is mostly well-differentiated, so the overall prognosis is better. However, children who have extracapsular invasion, multiple lesions in bilateral thyroid, cervical lymph node metastasis and distant metastasis are at high risks and should be treated with comprehensive therapy that includes total thyroidectomy.

Objective To study the clinical effects of addition treatment of liver-kidney yin deficiency and postmenopausal osteoporosis with Liuwei Dihuang pill.Methods 120 patients suffering from postmenopausal osteopo-rosis who were determined to be subject to liver -kidney Yin deficiency by doctors of traditional Chinese medicine when they received treatment at the outpatient clinic of traditional Chinese medicine department of our hospital in the period from February 2010 to june 2013 were divided into three groups:group A,B and C,with the random number table.For group A:provided the honeyed pills made up by supplemented six -ingredient rehmannia pill,and taken two pills per day continuously for six months;For group B:provided 5mg of zoledronic acid injection for intravenous drip infusion;For group C:provided traditional Chinese medicine honeyed pills and zoledronic acid for treatment at the same time;all of the three groups take Calcium carbonate D3 at the same time;The three groups of cases until the 12th month were observed and followed -up.The clinical efficacy of three groups were compared.Results A,C group 3 months after treatment with estradiol levels compared with before treatment,the difference was statistically significant (t =2.54,P =0.05,t =2.07,P =0.06).A group 3 months after treatment,the VAS scores at 6 months respectively (3.75 ±1.32),(2.06 ±1.53)points before treatment,and the differences were statistically significant(t =10.18, P =0.00,t =9.26,P =0.00).A group 3 months after treatment,the SF -36 scores at 6 months were (90.82 ± 12.38),(105.82 ±9.06)% respectively,compared with that before treatment,the differences were statistically significant(t =8.40,P =0.00,t =5.21,P =0.00).A group 6 months after treatment of lumbar 1 -4 bone density-were (0.85 ±0.07)g/m2 ,and before treatment[(0.76 ±0.07)g/m2 ]comparison,the difference was statistically significant(t =7.56,P =0.00);A group 6 months after treatment of Wards bone density compared with before treatment,the difference was statistically significant(t =8.07,P =0.00).C group 6 months after treatment of lumbar 1 -4 bone density(0.91 ±0.07)g/m2 ,and B Group[(0.88 ±0.05)g/m2 ]comparison,the difference was statistical-ly significant(t =27.19,P =0.00);C group after treatment,the bone mineral density of Wards area for 6 months were (0.62 ±0.04)g/m2 .The difference was statistically significant (t =26.22,P =0.00),compared with the B group. The TCMsyndromes of TCMin the 6 months after treatment in A group were (5.42 ±2.13),and the difference was statistically significant (t =13.29,P =0.00),compared with the B Group (10.75 ±4.08).(t =13.29,P =0.00). Compared with B group,the difference was statistically significant(t =26.22,P =0.00).A group after the treatment of TCMsyndrome score was 6 months(5.42 ±2.13),and B Group[(10.75 ±4.08)],the difference was statistically significant(t =13.29,P =0.00).Conclusion The addition of Liuwei Dihuang pill can improve BMD,alleviate the ostealgia and improve life quality of patients suffering from postmenopausal osteoporosis,and has no obvious adverse effects.It can enhance curative effects through using with diphosphonate.

OBJECTIVE:To observe the clinical efficacy and ADR of ultrasound debridement combined with Huangma tinc-ture in the treatment of diabetic foot ulcer. METHODS:90 diabetic foot ulcer patients were randomly divided into combination treatment group(ultrasound debridement combined with Huangma tincture wet compress),Huangma tincture group(Huangma tinc-ture wet compress) and control group (vaseline) with 30 cases in each group. 3 months after treatment,Ulcer healing rate,cure time,effective time,recovery time,cure rate and ADR were observed in each group. RESULTS:The ulcer healing rate of combi-nation treatment group was significantly higher than that of Huangma tincture group and control group,with statistical significance (P<0.05);there was statistical significance between Huangma tincture group and control group(P<0.05). Mean cure time,effec-tive time and recovery time of combination treatment group were all significantly lower than Huangma tincture group and control group,with statistical significance(P<0.05);there was statistical significance between Huangma tincture group and control group (P<0.05). Cure rate of combination treatment group was higher than that of Huangma tincture group and control group,with statis-tical significance (P<0.05);there was statistical significance between Huangma tincture group and control group (P<0.05). No ADR was found in combination treatment group and Huangma tincture group. CONCLUSIONS:The ultrasound debridement com-bined with Huangma tincture in the treatment of diabetic foot ulcer improve healing rate and shorten healing time significantly with-out obvious ADR. It has good clinical efficacy.

Aim To explore the effects of niacin on LDL-C uptake and metabolism in HepG2 cells,and to clarify the functions of niacin in lipid-lowering and slo-wing the atherosclerosis process,thus to provide a sci-entific basis for niacin as a lipid-lowering drug in clini-cal development.Methods Oil red O staining was used to observe HepG2 cells after lipid uptake.Enzy-matic method was used to determine the content of in-tracellular free cholesterol (FC)and total cholesterol (TC).The LDLR levels on the surface of cell mem-brane were detected by immunofluorescence flow cy-tometer.The mRNA and protein expressions of LDLR, SREBP2 and PCSK9 were analyzed by qPCR and Western blot.Results The results of oil red O staining showed that the rate of oil red O-positive cells and the number of red lipid droplets were significantly in-creased in niacin group than control group.Niacin sig-nificantly increased the levels of TC and FC in HepG2 cells(P <0.05 ).What’s more,niacin significantly upregulated the expression of LDLR and significantly downregulated the protein expression of PCSK9,while it had no effect on the expression of SREBP2.Conclu-sion Niacin accelerates LDL-C uptake probably via downregulating the expression of PCSK9 and reducing the degradation of LDLR protein in HepG2 cells.

AIM:To investigate high-density lipoprotein ( HDL) subclass distribution and to analyze the rela-tionship between HDL subclasses with plasma glucose and lipids in metabolic syndrome ( MS) .METHODS:Apolipopro-tein A-I ( apoA-I) contents of plasma HDL subclasses were determined by two-dimensional gel electrophoresis associated with immunodetection .The concentrations of lipids and apolipoproteins in the plasma were measured by an automated bio -chemical analyzer .RESULTS:Compared with the controls , the levels of fasting plasma glucose ( FPG) , total cholesterol (TC), triglyceride ( TG), low-density lipoprotein cholesterol ( LDL-C), LDL-C/high-density lipoprotein cholesterol (HDL-C), apolipoprotein B100(apoB100), apoB100/apoA-I, systolic blood pressure (SBP), body mass index (BMI) and HDL3b were increased in the MS patients (P<0.05).Meanwhile, HDL-C, apoA-I and preβ2-HDL, HDL2a and HDL2b were decreased in the MS patients (P<0.01).With the increase in the plasma glucose level , the contents of HDL2a and HDL2b were decreased in the MS patients (P<0.05), while preβ1-HDL was increased (P<0.05).With the decrease in the HDL-C level, the content of HDL2b was decreased in the MS patients (P<0.01), while preβ1-HDL was increased (P<0.01).With the increase in the TG level and the decrease in the HDL-C level, the content of HDL2b had a decrea-sing trend and the content of small-particle preβ1-HDL had an increasing trend , indicating that HDL maturation metabolism was disrupted.The correlation analysis showed that FPG was negatively correlated with the levels of HDL 2a and HDL2b, HDL-C was negatively correlated with the level of preβ1-HDL and positively correlated with the level of HDL 2b , and TG was positively correlated with the levels of preβ1-HDL and HDL3b .CONCLUSION:With the increases in the plasma glucose and TG, and the decrease in HDL-C in the MS patients, HDL particles have minifying tendency , and the maturation me-tabolism of HDL particles is disrupted .