Objective To investigate two fixation methods for reconstruction of anterior cruciate ligament(ACL)with autogenous patellar tendon graft and their instant stability.Methods 15 specimens were divided into three groups.The first group was treated with 1/3 bone-patellar ligament-bone replacement and in terference screw fixation,the second group was treated with 1/3 bone-patellar ligament and Krackow locked bilateral suture,and in the third group the intact ACL was not treated.The mono-axial tensile tests were performed on a versatile hydraulic material testing machine(WE-5A)to compare the linear load,maximum load,linear straining,maximum straining,linear displacement,maximum displacement,rupture displacement,destructive energy,elastic modulus,and ultimate strength of ACL in the three groups.Results The ACL stability and rigidity in the group of 1/3 bone-patellar-bone replacement and interference screw fixation were higher than those in the other two groups.The differences between test results were statistically significant.Conclusion The ACL reconstruction with 1/3 bone-patellar ligament-bone replacement and interference screw fixation is a good method to ensure higher strength of graft and better joint stability.

Graphene oxide (GO) modified stationary phase for open tubular capillary (3 m × 25 μm i. d. ) of liquid chromatography ( OT-CLC) was fabricated by coating GO sheets onto poly ( vinylbenzyl chloride-divinylbenzene) porous composite layer via covalent coupling, which was prepared by in-situ polymerization. Scanning electron microscopy ( SEM), Raman spectroscopy and transmission electron microscopy ( TEM) were used to characterize the structure of the stationary phase. The results demonstrated that poly(vinylbenzyl chloride-divinylbenzene) porous composite layer had decentralized globular structure, and the GO sheets covered on the porous layer homogenously. The phase ratio and sample capacity were greatly improved due to the spherical polymer layer and the coverage of GO. Thus, alkyl benzenes, neutral polycyclic aromatic hydrocarbons, acidic and basic compounds could be well separated by using acetonitrile-water as eluent, while four nucleobases were completely separated by using acetonitrile-0. 02 mol/ L ammonium acetate as mobile phase. The run-to-run, day-to-day, and column-to-column reproducibilities were evaluated by calculating the relative standard deviations (RSDs, n =6) of the retention time of o-phenylenediamine, aniline and 2, 4, 6-trifluoroaniline, respectively. These RSD values were all in the range of 0. 3% -2. 0% .

Objective To evaluate the pathological,histochemical and clinical features of primary clear cell carcinoma of the epididymis,and to investigate its diagnosis,treatment and prognosis.Methods The clinical data of one case of primary clear cell carcinoma of the epididymis were reported and the relevant literature was reviewed.Results The patient was a 43-year-old man presenting with a painless mass in the left epididymis.On physical examination,a 1.0 cm-sized smooth solid nodule on the head of left epididymis could be palpated without tenderness.Then the patient underwent left epididymectomy.On postoperative histopathological study,the tumor was characterized by diffuse sheets of clear cells and adenoid structures.Immunohistochemical assay showed AE(++),AE3(+/-),NSE(-) and interstitialoma(-) in the tumor tissues.Postoperatively,MRI showed no space-occupying lesion in both kidneys,and B-ultrasound indicated no abnormality of the bladder.Fifteen days after operation,left orchiectomy was performed on the patient.Pathology showed no clear cell carcinoma tissue of the testis.Follow-up was 15 months.The patient was reported to be alive and tumor-free.Conclusions The histogenesis of primary clear cell carcinoma of epididymis is unclear yet.The diagnosis depends on pathologic studies,and should be differentiated from nephrogenic adenoma and metastatic carcinoma.Radical surgery is the main treatment choice.

Objective: To express the protein of HPV18E6 based on pET-32a(+) at high level and study the expression and significance of HPV18E6 proteins in laryngeal carcinoma. Methods: The HPV18E6 gene was amplified by PCR and cloned into pET-32a(+). The amplified fragment was inserted into the plasmid pET32a (+) that was digested with BamHⅠand Hind Ⅲ. The recombinant plasmid pET32/E6 was transformed into E.coli JM109 which was selected with ampicillin. The recombinant plasmids were successfully introduced into E.coli BL21(DE 3) and were induced by IPTG. SDS-PAGE and Western blot analysis were used to detect the confusion protein. Finally, the optimization of expression conditions, such as temperature, concentration of IPTG, was studied. Results: The recombinant plasmids were identified and confirmed with enzyme digestion and sequencing. The BL21(DE3) transformed recombinant plasmid pET32/E6 had expressed HPV18E6 recombinant protein effectively. The optimum conditions of expression were 37 ℃, 1 mmol/L IPTG. Conclusion:Prokaryotic expression vector pET-32a(+)-HPV18E6 was successfully constructed. The high-level expression of HPV18E6 was achieved in E.coli BL21(DE3).

Objective To evaluate whether low carbohydrate diet before 18F-FDG tumor imaging could reduce myocardial 18F-FDG uptake.Methods From April 2011 to January 2012,70 patients were enrolled in this study.They were randomly divided into control group (34 cases) and test group (36 cases).Patients in control group were on regular diet,while those in test group had low carbohydrate diet in the evening before imaging.Blood samples were taken before injection of 18F-FDG for the measurement of serum glucose,free fatty acid,insulin and ketone body.Whole body 18F-FDG tomography was performed with dualhead coincidence SPECT.The myocardial uptake of FDG was assessed visually and scored as 0 for no uptake,1 for uptake lower than liver,2 for uptake similar to liver,3 for uptake higher than liver,and 4 for remarkable uptake.The ratio of myocardium to liver (H/L) was calculated.Two-sample t test,Wilcoxon rank sum test and linear correlation analysis were performed.Results The myocardial uptake in test group was significantly lower than that in control group with H/L ratios of 0.94±0.57 and 1.50±1.04,respectively(t=-2.75,P＜0.05).The concentrations of serum free fatty acid and ketone body in test group were significantly higher than those in control group: (0.671±0.229) mmol/L vs (0.547±0.207) mmol/L and (0.88±0.60) mmol/L vs (0.57±0.32) mmol/L,t=2.38 and 2.67,both P＜0.05.The concentrations of glucose and insulin were (5.28±1.06) mmol/L and (35.16±33.70) pmol/L in test group,which showed no significant difference with those in control group ((5.19±0.78) mmol/L and (41.64±35.13) pmol/L,t=0.39 and-0.79,both P＞0.05).A negative correlation was found between the myocardial uptake of 18F-FDG and serum free fatty acid/ketone body concentration (r=-0.40,-0.33,both P＜0.01),respectively.There was no correlation between the myocardial uptake of 18 F-FDG and glucose/insulin (r =-0.02,0.13,both P＞0.05),respectively.Conclusion Low carbohydrate diet before 18F-FDG tumor imaging can reduce myocardial uptake,thus facilitating detection of lesions near the heart.

Objective To analyse the effect of holmium laser incision through ureteroscopy and simple ureteroscopy treatment for refrac-tory hemospermia. Methods From December 2003 to April 2013,the data of 67 cases with refractory hemospermia were retrospectively ana-lyzed. All the patients underwent semen analysis,transrectal ultrasonography,seminal vesicle ultrasonography,some patients underwent pelvic CT or MRI. Results Simple ureteroscopy were done for 24 cases,holmium laser incision through ureteroscopy were done for 43 cases. Var-ying degrees of ejaculatory duct stenosis or obstruction were observed. Postoperative follow-up was from 6 months to 8 years,in 24 cases of simple ureteroscopy,2 cases experienced recurrence 6 or 8 months later. The ejaculatory duct narrow were found when they received reopera-tion,with holmium laser incision,hemospermia disappeared. No complications such as retrograde ejaculation,urinary incontinence or rectal injury occurred postoperatively. Conclusion The effect of holmium laser incision through ureteroscopy for refractory hemospermia is better than simple ureteroscopy,which is worthy of clinical application needs further observation and summary.

Aim To investigate the modeling of breast cancer in zebrafish embryos and its related protein expression. Methods 48hpf wild type AB/ TG(Transgenic) zebrafishs were micro-in-jected with breast cancer cell line: MCF-7,T-47D, MDA-MB-231 respectively, the relationship between the number of tumor and model application was investigated, and the number of sub-intestinal veins(SIVs) was detected under confocal microscope, as well as the metastasis of tumor cells in embryos; then the ze-brafish xenografts of MB-231 were co-cultured with tofacitinib/ptk787 for 48 h, optical density(OD) of the cell survival and subintestinal veins(SIVs) were evaluated under confocal micro-scope, and Western blot(WB) analysis was used to test micro-circumstances related protein. Results When the number of in-oculated cells was more than 200 per embryo, xenograft model rate woule be more than 0. 90;MB-231 xenografts showed metas-tasis feature in zebrafish, which could be inhibited by tofacitinib (P < 0. 01), while the number of xenograft MB-231 cells was reduced significantly(P < 0. 01); in another zebrafish xenografts SIVs assay, the tumor could promote the proliferation of SIVs, and 4 mg·L - 1 PTK787 showed inhibiton effect( P < 0. 01). Western blot showed 4d T-47D xenograft zebrafish got more HER2 expression than AB embryos; VEGFa expression in ze-brafish MB-231 model group was higher, and model zebrafish P53 expressi was higher after treated by tofacitinib. Conclusion A zebrafish xenograft model of human brest cancer can be es-tablished, which demonstrates applicability for screening com-pounds in drug discovery studies.

Prokaryotic expression vector of mouse HPV16E6 gene was constructed. A pair of primers were designed according to the digestion sites in plasmid pGEX-KG and the HPV16E6 gene sequence published by GenBank. The DNA fragment of 321bp was amplified by PCR from the HPV recombinant plasmid with HPV16E6 gene, then cloned into pGEX-KG and transformed into the host E. coli strain JM109. The fragment was conformed to the original sequence, which indicated that fusion expression vector pGEX-KG-HPV16E6 was constructed. The pGEX-KG-HPV16E6 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degrees C, the expression product of HPV16E6 gene was identified by SDS-PAGE and Western blot. HPV16E6 fusion protein had been expressed successfully in the form of inclusion bodies, the molecular weight of fusion protein being 38 kD. Meanwhile, the optimum condition of HPV16E6 fusion protein expression was induced with 1.0 mmol/L IPTG for 4h. The fusion protein reacted specifically with the antibodies against HPV16E6. HPV16E6 gene was successfully expressed in E. coli, which could be used as a basis for preparing HPV16E6 vaccine in human.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Repressor Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology

Objective To evaluate the clinical effect of milrinone nebulized inhalation for improving intraoperative cardiac function and pulmonary arterial pressure in the patients with chronic obstructive pulmonary disease (COPD) complicating pulmonary hypertension (PH).Methods Forty-four surgical patients with COPD complicating PH in the Chongqing Municipal Medical Emergency Center from June 2015 to June 2016 were chosen,including 23 cases of thoracic surgery,13 cases of abdominal surgery and 8 cases of lower extremity fracture surgery.The patients were divided into the control group and treatment group,22 cases in each group.The control group received the routine comprehensive treatment.In addition receiving the conventional comprehensive treatment,milrinone nebulized inhalation in the treatment group was given before general anesthesia.Both of the two groups were imbedded with floating catheter in the right internal jugular vein for detecting the clinical indexes:cardiac output (CO),pulmonary artery systolic pressure (PASP),pulmonary artery mean pressure (PAMP) and pulmonary capillary wedge pressure (PCWP).Results Compared with before treatment,CO after treatment in the treatment group was significantly increased (P＜0.05),PASP,PAMP and PCWP after treatment in the treatment group were significantly decreased(P＜0.05).CO,PASP,PAMP and PCWP in the control group had no statistical difference between before and after treatment,the difference was not statistically significant (P＞0.05).Conclusion Intraoperative milrinone nebulized inhalation in the patients with COPD complicating PH can effectively improve the patient's cardiopulmonary function.

Aim To investigate the anti-angiogenesis and anti-xenograftes of UA in zebrafish larvaes. Meth-ods 24 hour post-fertilization ( hpf ) TG zebrafish was treated with different concentration of UA for 24h, and the number of intersegmental vessels( IVS) were detec-ted under fluorescent microscope respectively;then the models of AB/TG zebrafish xenografts were established by be micro-injected with SMMC-7721 or HT-29 cell at 48hpf respectively, and after cocultured with UA for 48h, optical density (OD) of the SMMC-7721 cell and subintestinal veins ( SIVs) induced by HT-29 were e-valuated under confocal microscope. Results ISV as-say showed that UA could cause IVS missing or disap-perance, inhibition ratio reaching 20. 25% and 26. 65%. UA blocked the spread of SMMC-7721 cells in zebrafish and OD value,and inhibition ratios at three concentrations were 38. 01%, 54. 69%, 61. 88%, re-spectively; in another SIVs assay induced by xeno-grafts, UA at concentration 10 and 15mg·L-1 showed that SIVs were inhibited (P<0. 01) obviously. Con-clusion UA could inhibit the angiogenesis and the growth of SMMC-7721/HT-29 xenografts,and the anti-tumor mechanism may be related with VEGFR2 expres-sion.