Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

BACKGROUNDVascular control and tissue dissection are crucial steps in successful laparoscopic surgery. Recently, a new commercially available vessel sealing technology, the LigaSure vessel sealing system (Valleylab, Boulder, USA), has been introduced. The aim of the present study was to evaluate the benefits of the LigaSure in laparoscopic nephrectomy.

METHODSFrom January 2005 to March 2010, 170 laparoscopic nephrectomies were performed with the LigaSure vessel sealing system, including simple and radical nephrectomy and nephroureterectomy. In a retrospective study, the laparoscopic operating time, estimated intraoperative blood loss, duration of postoperative drainage, total amount of postoperative drainage, as well as postoperative hospital stay, were recorded and studied.

RESULTSAll 170 laparoscopic nephrectomies using LigaSure were accomplished successfully without conversion to open surgery. There was no severe vascular complication or other serious complications. The mean laparoscopic operating time was 124.2 minutes (range, 14 - 230 minutes); mean blood loss was 148.6 ml (range, 20 - 540 ml); mean time for postoperative drainage was 3.1 days (range, 1 - 7 days); mean amount of postoperative drainage was 206.5 ml (range, 27 - 435 ml) and mean postoperative hospital stay was 6.9 days (range, 3 - 18 days).

CONCLUSIONSLaparoscopic nephrectomy using LigaSure appears technically feasible and easy, and produces satisfactory results. The LigaSure provides a safe and fast way to seal vessels and tissue bundles during nephrectomy.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Nephrectomy ; methods ; Retrospective Studies ; Young Adult

OBJECTIVETo construct a short hairpin RNA (shRNA) vector of the hypoxia inducible factor-1alpha (HIF-1alpha), determine its inhibitory effect on the expression of the HIF-1alpha gene in PC-3M cells, and investigate its application prospects in the treatment of prostate cancer.

METHODSWe designed and synthesized the shRNA template sequence specific against HIF-lalpha, inserted it into the vector psilencer 2.1-U6 to generate the plasmid psilencer-HIF, transfected the recombinant plasmid into prostate cancer cell line PC-3M cells and detected the transfection efficiency by cotransfection with the pEGFP vector as well as the expression of HIF-1alpha by RT-PCR and Western blot.

RESULTSThe DNA sequencing analysis showed a complete consistency of the recombinant plasmid psilencer-HIF with the design. Twenty-four hours after the transfection, the rate of transfected plasmid was about (89.26 +/- 4.72)% and the vector-mediated shRNA induced RNA interference (RNAi), while 48 hours transfection reduced the HIF-1alpha mRNA and protein levels by 82.09% and 81.61% respectively (P < 0.01) in PC-3M cells.

CONCLUSIONThe shRNA vector was successfully constructed, which can effectively suppress the expression of HIF-1alpha in prostate cancer cells.

Base Sequence ; Cell Line, Tumor ; Gene Expression ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Male ; Prostatic Neoplasms ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Transfection

OBJECTIVETyping of Mycobacterium tuberculosis strains and epidemiological studies in the army of southern China to provide scientific basis for prevention of pulmonary tuberculosis.

METHODSA rapid fingerprinting of M. tuberculosis strains method by polymerase chain reaction (PCR) with outward-directed primers that designed to the ends of the insertion sequence IS6110 was developed, and to analyze the relationship between the polymorphism of DNA fingerprinting and epidemiology of M. tuberculosis.

RESULTSOne hundred and fifty-four M. tuberculosis detected were classified into eight types according to their characters of PCR amplified fingerprints. The main types were type I (36.4%), type II (31.8%), and type III (21.4%), while other types were less than 4 percentage. In those main type groups, patients aged 20 to 29 and 30 to 39 took up 31.8% and 27.9% respectively. For those main types, the distribution of those types in the first treated patients showed significant difference compared with that in the retreated patients, and the rate of drug-resistance was also statistically different. However, the distribution was not statistically significant to history of BCG vaccination and patients living in urban or rural area. The main drug-resistant strains were only Isoniazid-resistant or Rifampin-resistant strains, while the drug-resistant strains were 44.4%, 29.6% and 14.8% respectively in type I, type II and type III.

CONCLUSIONPCR fingerprinting was a rapid, precise, sensitive, specific method to type M. tuberculosis, and could be used to study the epidemiology of tuberculosis; The prevalence of tuberculosis was primarily due to the transmission of type I, type II and type III in the army being studied from Southern China, to suggest that surveillance needs to be strengthened.

Adult ; China ; epidemiology ; DNA Fingerprinting ; methods ; DNA, Bacterial ; genetics ; Female ; Humans ; Male ; Military Personnel ; Molecular Epidemiology ; Mycobacterium tuberculosis ; classification ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Sensitivity and Specificity ; Tuberculosis ; epidemiology ; Tuberculosis, Multidrug-Resistant ; epidemiology ; microbiology

Adult ; Aged ; Endoscopy ; methods ; Female ; Humans ; Kidney Calculi ; surgery ; Male ; Middle Aged ; Nephrectomy ; methods ; Postoperative Complications ; Retrospective Studies ; Treatment Outcome

BACKGROUNDPercutaneous nephrolithotomy (PCNL) is the most widely recommended treatment for calyceal diverticular calculi, providing excellent stone-free results. However, its invasiveness is not negligible considering its major complication rates. Flexible ureteroscopy (FURS) is currently used to treat calyceal diverticula. However, the greatest drawback of FURS is locating the diverticulum since its neck is narrow and concealed. In such a case, the FURS procedure must be converted to PCNL. The aim of this study was to evaluate ultrasound-guided flexible ureteroscopy (UFURS) identifying diverticulum and the management of calyceal diverticular calculi.

METHODSA retrospective analysis was conducted on 24 patients who had calyceal diverticular calculi. In all 12 patients in the UFURS group, direct FURS failed to find evidence of calyceal diverticula but were confirmed with imaging. The other 12 patients in the PCNL group received PCNL plus fulguration of the diverticular walls.

RESULTSPuncture of calyceal diverticulum was successful in all 12 UFURS patients. Two patients in this group had postoperative residual calculi and two patients developed fever. In the PCNL group, percutaneous renal access and lithotomy were successful in all 12 patients. One patient in this group had residual calculi, one had perirenal hematoma, and two patients developed fever. No significant difference was found in the operating time (UFURS vs. PCNL, 91.8 ± 24.2 vs. 86.3 ± 18.7 min), stone-free rate (UFURS vs. PCNL, 9/12 vs. 10/12), and rate of successful lithotripsy (UFURS vs. PCNL, 10/12 vs. 11/12) between the two groups (all P> 0.05). Postoperative pain scores in the FURS group were significantly lower than that in the PCNL group (2.7 ± 1.2 vs. 6.2 ± 1.5, P< 0.05). Hospital stay in the UFURS group was significantly shorter than that in the PCNL group (3.4 ± 0.8 vs. 5.4 ± 1.0 days, P< 0.05). All patients were symptom-free following surgery (UFURS vs. PCNL, 10/10 vs. 12/12).

CONCLUSIONUltrasound-guided puncture facilitates identification of calyceal diverticula during FURS and improves the success rate of FURS surgery.

Adult ; Diverticulum ; diagnosis ; surgery ; Female ; Humans ; Kidney Calculi ; diagnosis ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Ultrasonography ; methods ; Ureteroscopes ; Ureteroscopy ; instrumentation ; methods

BackgroundRobot-assisted/laparoscopic intracorporeal ileal conduit (ICIC) has been reported in many experienced centers. Whether laparoscopic ICIC is superior to extracorporeal ileal conduit (ECIC) and whether laparoscopic ICIC should be promoted is still controversial. The aim of the study was to compare surgical and early oncological outcomes between patients undergoing laparoscopic radical cystectomy (LRC) with ICIC and ECIC.

MethodsFrom January 2011 to June 2016, a total of 45 patients with bladder cancer underwent LRC with ileal conduit at our department, of whom 20 patients underwent LRC with ECIC and 25 patients underwent LRC with ICIC. Data of each patient's characteristics, surgical outcomes, and short-term oncological outcomes were collected and analyzed.

ResultsLRC with ileal conduit was performed successfully on all 45 patients. There were no significant differences in patients' characteristics, mean total operative time, and mean estimated blood loss between the ICIC and ECIC groups. Median time of flatus and oral intake was shorter in the ICIC group compared with the ECIC group (3 vs. 5 days, P = 0.035; 4 vs. 5 days, P = 0.002). The complications rates did not show significant difference between the two groups within the first 90 days postoperatively (P = 0.538). Cancer staging showed 45% of patients in the ECIC group and 36% in the ICIC group had a pathologic stage of T3 or T4, and 50% of patients in the ECIC group and 44% in the ICIC group had a pathologic stage of N1 or N1+. Kaplan-Meier analysis showed no significant difference in overall survival at 24 months (60% vs. 62%, P = 0.857) between the ECIC and ICIC groups.

ConclusionsICIC after LRC may be successful with the benefits of faster recovery time. No significant difference was found in complications and oncological outcomes between ICIC and ECIC. However, larger series with longer follow-up are needed to validate this procedure.

Aged ; Cystectomy ; methods ; Female ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Urinary Bladder ; surgery ; Urinary Bladder Neoplasms ; surgery ; Urinary Diversion ; methods

OBJECTIVE To stud y the chemical cons tituents of n-butanol part of Qubai tablet and its pharmacodynamic effect on the model of de melanocyte. METHODS The n-butanol part of Qubai tablet was prepared. The chemical constituents were analyzed by ultra-high performance liquid chromatography-mass spectrometry （UPLC-MS）. Taking mice B 16 melanoma cells as the research object ，the de melanocyte model was established and divided into model group ，positive control different concentration groups（8-methoxypsoralen 10，50，100，150，200 μmol/L），solvent group （diluted with DMSO ）and Qubai tablet n-butanol part different concentration groups （10，50，100，150，200 μmol/L）. The number of cells were observed by inverted microscope ，and the cell proliferation rate ，the rate of melanin production and promotion rate of tyrosinase activity were also detected. RESULTS In the positive and negative ion mode ，53 compounds in the n-butanol part of Qubai tablet were preliminarily determined （29 in the positive ion mode ，33 in the negative ion mode ，overlapping 9），of which coumarins accounted for the largest proportion ， followed by flavonoids. The n-butanol part of Qubai tablet could significantly increase the number of cells ，which was positively correlated with the action time and administration concentration. It could significantly increase the proliferation rate of cells ，the rate of melanin production and promotion rate of tyrosinase activity （P＜0.01）. CONCLUSIONS Coumarins and flavonoids may be the material basis for the anti-vitiligo effect of n-butanol part from Qubai tablet ；anti-vitiligo effect of n-butanol part of Qubai tablet may be realized by promoting tyrosinase activity.

Objective To investigate the clinicopathological features and prognosis of chromophobe renal cell carcinoma(ChRCC). Methods The clinical and pathological data of 126 patients with ChRCC treated in Cancer Hospital of Chinese Academy of Medical Sciences were retrospectively analyzed. Results The patients included 64 males and 62 females,with the age of 22-80 years(median of 52 years).The tumor was located on the right side in 70 cases and on the left side in 56 cases.Ultrasound,CT or magnetic resonance imaging(MRI)were performed.Of the 110 cases receiving ultrasound examination,63,23,13,10,and 1 cases showed hypoecho,hyperecho,isoecho,uneven or mixed echo,and dark hypoecho,respectively.Color Doppler flow imaging showed no blood flow signal in 42 cases and low blood flow signal in 60 cases out of 68 cases with blood flow signal.Among the 54 cases receiving CT,50 cases showed equal density or low density and 4 cases showed high density with clear boundary.The enhanced scanning showed mild to moderate uniform or non-uniform reinforcement,mostly below the renal parenchyma,and still showed reinforcement in the delayed period.Among the 97 cases receiving MRI,96 cases showed hypo-or isointense signals and 1 case showed hyperintense signal in T1 weighted images;71 cases showed hyper-or isointense signals and 26 cases showed hypo-or isointense signals in T2 weighted images;93 cases showed hyperintense signals with obvious limited diffusion and 4 cases showed unobvious limited diffusion in diffusion weighted images.Mild to moderate uniform or non-uniform reinforcement was observed in most of the enhanced scans.All the 126 patients underwent surgical treatment,including 64 cases of nephron sparing surgery and 62 cases of radical surgery.Pathological examinations confirmed ChRCC for all the patients,including 91 cases of T1N0M0,15 cases of T2N0M0,and 20 cases of T3N0M0.The immunohistochemical assay demonstrated the positive expression rate of 48.2%(54/112)for CD10,92.3%(96/104)for CD117,8.0%(9/112)for vimentin,85.6%(95/111)for CK7,and 97.6%(83/85)for colloidal iron.Conclusions ChRCC is less common,with low level of malignancy and good prognosis.Since the clinical symptoms of ChRCC are not typical,MRI is an important means of imaging differential diagnosis,and the disease can be confirmed depending on pathological diagnosis.Surgery is the preferred treatment method,and currently there is no standard treatment regimen for metastatic patients.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Renal Cell/diagnostic imaging* ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Kidney Neoplasms/surgery* ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Prostate cancer (PC) is one of the malignant tumors of the genitourinary system that occurs more often in elderly men. Screening, early diagnosis, and treatment of the PC high risk population are essential to improve the cure rate of PC. The development of the guideline for PC screening and early detection in line with epidemic characteristics of PC in China will greatly promote the homogeneity and quality of PC screening. This guideline was commissioned by the Bureau of Disease Control and Prevention of the National Health Commission. The National Cancer Center of China initiated and convened a working group comprising multidisciplinary experts. This guideline strictly followed the World Health Organization Handbook for Guideline Development and combined the most up-to-date evidence of PC screening, China's national conditions, and practical experience in cancer screening. A total of fifteen detailed evidence-based recommendations were provided with respect to the screening population, technology, procedure management, and quality control in the process of PC screening. This guideline aimed to standardize the practice of PC screening and improve the effectiveness and efficiency of PC prevention and control in China.
Aged ; Beijing ; China/epidemiology* ; Early Detection of Cancer ; Humans ; Male ; Mass Screening ; Prostatic Neoplasms/epidemiology*