The diagnosis of polypoid lesions of the gallbladder plays a key role in the selection of treatment methods.However we still lack of effective means to exclude the malignant transformation currently.It is believed that lesions,whose diameter more than 10 mm,tend to be malignant.In addition,multiple imagings and related risk factors can also bring us some valuable tips.This article summarizes the current research progress related to the differential diagnosis of the disease,hoping to provide help for the clinicians in the timing of operation for such disease.

Objective To establish a reliable approach for quantification of colony forming unit(CFU)of Mycobac-terium tuberculosis (M.tb)by measuring optical density(OD).Methods M.tb suspension H37Ra was prepared using low-power ultrasonic or glass bead beating methods,and was two-fold serially diluted,OD at 600nm (OD600)of each dilution ratio was measured respectively,OD600 and dilution curve were analyzed to determine the optimum approach for preparing bacterial suspension,linear range of OD600,as well as linear relationship between OD600 and CFU.Results OD600 was 0.1 -0.6,linear regression analysis of OD600 and dilution ratio within linear range revealed that correlation coefficient (R2 )of glass bead beating and low-power ultrasonic methods were 0.98 and 1 .00 respectively,both presented a good correlation,low-power ultrasonic method was better than glass bead beat-ing method,bacterial suspension dispersed more evenly.Linear regression analysis results of OD600 and CFU val-ues showed that the regression equation of glass bead beating method and low-power ultrasonic method were CFU=2.35×107 ×OD600+4.42×105 and CFU=3.26×107 ×OD600+6.89×105 respectively.Conclusion Low-power ultrasonic method is a good method for preparation of M.tb suspension,combined the measurement of OD600 value, it can be a reliable and rapid method for quantitative analysis of M.tb.

High-speed counter-current chromatographic ( HSCCC ) method was successfully used to separate and purify prim-O-glucosylcimifugin and 5-0-methylvisammioside from Saposhnikovia divaricata(Thicz. ) Schis-chk. ,a traditional Chinese herb,with a solvent system composed of ethyl acetate-n-butanol-water(2:7:9,V/V). The lower phase of the system was used as the mobile phase at the flow rate of 2.0 mL/min,and the upper phase was used as the stationary phase. The separation produced a total 13.9 mg prim-O-glucosylcimifugin and 25.0 mg 5-0-methylvisammioside with the purity of 98. 1% and 99.2% determined by high performance liquid chromatography (HPLC) from 316 mg of crude sample from S. divaricata. The structures of isolated compounds were identified by ESI-MS,~1H NMR and ~(13)C NMR.

BACKGROUND:Cationic liposome-mediated celltransfection is reliable and repeatable. However the transfection efficiency is often low.
OBJECTIVE:To study the optimized methods for gene transfection mediated by liposome into N2a cells (mouse neuroblastma cells).
METHODS:Using traditional adherent method and improved suspension method, 500 ng recombinant plasmid pcDNA3-GFP carrying green fluorescence protein was transfected into N2a cells in 24-wel culture plate, which was mediated by 1.5μL Lipofectamine?LTX Reagent. The expression of green fluorescent protein was observed by inverted fluorescence microscope, and the transfection efficiencies at different transfection ways were calculated. By using improved suspension transfection method, 500 ng plasmid DNA was transfected with different doses of Lipofectamine?LTX Reagent (1.0, 1.5, 2.0, 2.5μL). The optimal ratio of liposome and DNA was explored.
RESULTS AND CONCLUSION:The transfection efficiency of suspension transfection method was significantly higher than that of the tranditional adherent method (P<0.01) when using 1.5μL liposome/500 ng DNA. The transfection efficiency of the 1.0, 1.5, 2.0, 2.5μL Lipofectamine?LTX on 500 ng plasmid DNA was respectively (76.60±3.85)%, (80.00±4.17)%, (88.00±5.89)%, (54.96±4.23)%. It showed the 500 ng DNA and 2.0μL liposome achieve the highest transfection efficiency.

The insecurity factors of third-class steel implants in the clinical application are introduced.A method for organizing the stocking way and improving the management in hospital are used to monitor and manage all the processes of the steel implants.All problems in management are solved,and this can insure the operation security and depress the costs of patients.

Objective To evaluate the effect of external direct current shock on left atrial(LA) diamension and volumes after cardioversion for atrial fibrillation(AF) and the relation between LA size and function.Methods The initial study population included 68 patients with AF.According to cardioversion patterns, the patients were divided randomly into cardioversion with direct-current shock(36 patients) and cardioversion with drugs(32 patients). Echocardiographic evaluations included LA size and volume,LA passive and active emptying volumes were calculated,and LA function was measured as LA ejection force (LAEF).The correlation between LA diameters and volumes and LA systolic function was analyzed. Results LA was dilated in all the patients during AF and the superior-infra dimensions of LA decreased after restoration of sinus rhythm in the patients reverted with external direct current shock and the patients reverted with durgs (P

Objective To analyze the safety and effectiveness of ambulatory surgery in inguinal herniorraphy.Methods The clinical data of 3 852 cases of inguinal hernia repair patients admitted from January 2009 to December 2013 in this single center was analyzed retrospectively.Cases of emergency surgery were excluded.Results All patients had long-term follow-up (12-60 months).1 575 patients underwent day-surgery,mean operation time was (43.84 ± 12.35) min,mean time of ambulation was (1.12 ± 0.91) d,mean time of recovery was (5.78 ± 1.12) d,mean hospitalization was (1.34 ± 0.48) d,mean hospitalization cost was (7 546.49 ±2 962.57) RMB.In contrast,there were 511 patients underwent a non day-surgery,mean operation time was (48.59 ± 14.52) min,mean time of ambulation was (2.43 ± 1.38) d,mean time of recovery was (7.46 ± 2.62) d,mean hospitalization was (4.8 ± 2.91) d,mean hospitalization cost was (9 165.16 ± 4 281.83) RMB.Patients with day-surgery were significantly superior to those with non day-surgery in operation time (P =0.000),mean time of ambulation (P =0.000),mean hospitalization (9 =0.000),mean hospitalization cost (P =0.000) and mean time of recovery (P =0.000).Infection and readmission in non day-surgery patients was higher than that in day-surgery (P =0.000).There was no difference in postoperation pain and hernia recurrence.Conclusions Ambulatory surgery in inguinal herniorraphy is safe with similar recurrence rate;but significantly lower cost and shorter hospitalization.

Objective To investigate the surgical method and clinical efficacy of hilar cholangio carcinoma in Bismuth type Ⅳ.Methods The retrospective descriptive study was adopted.The clinical data of 1 patient with hilar cholangiocarcinoma in Bismuth type Ⅳ who was admitted to the Renji Hospital affiliated to Shanghai Jiaotong University in October 2014 were collected.The patient had complaint about right upper abdominal pain for half month.Enhanced CT scan showed soft-tissue mass at hepatic hilum.After accurate assessment,the patient underwent radical resection of hilar cholangiocarcinoma + right hemihepatectomy + perihilar resection + right caudate hepatectomy + Roux-en-Y hepaticojejunostomy.The operation time,volume of intraoperative blood loss,results of pathological examination,postoperative complications,time of drainage tube removal,discharge time and follow-up were observed.The follow-up was performed to detect the life quality and tumor recurrence by outpatient examination and telephone interview up to July 2015.Results The patient received successful radical resection of hilar cholangiocarcinoma + right hemihepatectomy + perihilar resection + right caudate hepatectomy + Roux-en-Y hepaticojejunostomy.Operation time and volume of intraoperative blood loss were 480 minutes and 300 mL,respectively.The result of pathological examination showed that the size of hilar bile duct was 4 cm× 3 cm × 2 cm and poor-differentiated adenocarcinoma infiltrated through bile duct into liver tissues and right branch of portal vein.Two lymph nodes in the 8th group,1 in the 12a group and 3 in the 12p group were positive by detection,showing the metastasis of cancer cells.The resection margins of liver and bile ducts were negative,achieving a R0 resection.The patient had a removal of negative pressure drainage tube at postoperative day 7 and discharged from hospital at postoperative day 12,with a good recovery and without the complications of biliary fistula,abdominal infection and hepatic failure.During the 9-month follow-up,there was a good life quality and no tumor recurrence.Conclusion The radical resection rate of Bismuth type Ⅳ tumor can be increased by accurate preoperative evaluation,rational surgical approach,individualized surgical planning and precise intraoperative procedures.

Objective To determine whether pulmonary arterial hypertension(PAH) and pulmonary venous hypertension (PVH) can be differentiated noninvasively by echocardiography. Methods Fifty-six patients with pulmonary arterial systolic pressure(PASP) ≥40 mmHg by echocardiography were involved,and cardiac catheterization performed within 7 days of each other. Based on left ventricular end-diastolic pressure or pulmonary capillary wedge pressure(PCWP) ,30 patients were classified as PAH group and 26 patients as PVH group. The early(E) and late(A) diastolic mitral inflow velocities,E/A ratios,deceleration time(DT),early dastlic mitrial annular velocity(E') and E/E' ratios were measured by conventional and Doppler tissue imaging echocardiography in the two groups. Results Compared with PVH group,the PAH group had significantly higher A,DT,PASP and E',and significantly lower E,E/A ratio and E/E' ratio (P < 0. 01 or P <0. 001). E/E' and E/A ratio was optimal indexes for differentiation of PAH and PVH,the area under receiver operating characteristic curve was 97% and 91 %, respectively. Optimal cutoff for diagnosing PVH was E/E'>9.2 (sensitivity 95%, specificity 97%) and E/A> 1. 7 (sensitivity 75%,specificity 92%). Conclusions PAH and PVH imaging can be reliably differentiated by echocardiography.

Objective To establish a long-term culture system for mouse spermatogonial stem cells(SSCs). Methods Testis cells from 4-6 days postpartum male transgenic BALB/C mice were collected by a modified two-step enzymatic digestion method.After three differential adherence selections,the enriched germ cells were finally suspended in StemPro-34 SFM medium supplemented with other nutrients factors and plated on mouse embryonic fibroblast(MEF)feeder layer.20 ng/ml Glial cell line-derived neurotrophic factor,10 ng/ml basic fibroblast growth factor and 200 ng/ml GDNF-family receptor al were added to the serum-free medium to promote SSCs proliferation.Aduh male BALB/C mice,4-5 weeks old,underwent intraperitoneal injection of 40 mg/kg busulfan as recipient mice.Cultured SSCs were also injected into the seminiferous tubules of the left recipient testis through micromanipulator and right testis as self-control.Testes of recipient mice were observed by a fluorescence stereomicroscope and HE stains at 2 months after transplantation. Results By improved digestion method,the vitality of isolated testis cells was more than 98%and the stem cells was enriched about 18.5 fold. 1-2 days after transferred to MEF feeder, the round germ cells started to proliferate and had the shape of paired or aligned undifferentiated spermatogonia connected by cytoplasmic bridges. After 3-4 days, SSCs proliferated continuously and formed typical colonies. SSCs from BALB/c mice could be cultured and passaged in a steady state for 3 months. Cryostat section through the transplanted testis showed that most of seminiferous tubules were filled with germ cells expressing EGFP.HE staining further showed clearly that seminiferous tubules contained complete spermatogenesis.Conclusions SSCs from BALB/c mice could be cultured in an improved culture system for 3 months.The culture system could facilitate understanding the regulatory mechanism that governs SSCs and might provide an opportunity for the cure of infertility.