Objective To investigate the efficacy and feasibility of extra-peritoneal larparoscopic radical prostatectomy (eLRP) in the treatment of patients with high-risk prostate cancer (HRPC).Methods From February 2009 to December 2013,121 patients,who were diagnosed as HRPC according to the D'Amico definition,were received eLRP.The mean age was 70 years old (range 54 ～ 82 years old).The mean PSA level was 25.45 (range 2.40 ～ 111.31) μg/L and mean Gleason score was 8 (range 6 ～ 10).The classification of clinic stage in this study included 52 cases in cT1-cT2b,58 cases in cT2c,8 cases in cT3a,and 3 cases incT3b,respectively.The perioperative data were collected,including operative time,blood loss,intraoperative complications,urine leakage,lymph leakage,incontinent ability,erectile function and changing of PSA level.Results All the operations were successfully performed.The mean operative time was 165 minutes (range 105 ～341min),the average blood loss was 150 ml(range 50 ～ 1500ml).The intraoperative complications included hemorrhage in 4 cases and intra-operative obturator nerve injury in 3 cases.The mean duration of intestinal function recovery was 35h (range 24 ～72h) The mean interval of catheter indwelling was 9 days (range 7 ～14 days).The anastomotic leakage was found in 12 cases,including 1 day after surgery in 5 cases,2 days after surgery in 3 cases,3 days after surgery in 2 cases,4 day after surgery in 1 case and 5 day after surgery in 1 case.The anastomotic stricture in 3 cases within 2 to 4 months after operation,which the symptom improved after urethral dilation in 2 cases and urethrotomy in 1 case.Deep vein thrombosis was noticed in 1 case 5 days after the procedure.And lymphatic fistula was recorded in 1 case after the operation.Positive surgical margin,seminal vesicle invasion,and positive iliac vessel lymph node were found in 18,21,and 9 patients,respectively.The mean hospitalization duration was 10 days (range 5 ～ 22 d).Of the 107 patients followed-up,Ninety-six patients were continent in 1 year,except other 11 patients.Nerve sparing procedure was performed in 51 patients,and thirty-three of them were potent.The mean PSA level was 0.14 μg/L (range 0 ～8.75 μg/L) six weeks after the surgey.Fourty-eight patients had biochemical recurrence with 5 ～36 months followed-up,mean 18 months.Conclusions Extraperitoneal LRP is an efficacious approach for patients with high-risk prostate cancer.

Background The establishment of diabetic animal model is a crucial step for the study about diabetic eye diseases. At present,the main modeling method include the injection of streptozotocin and alloxan. But the shortcoming of the former is an expensive price, and that of the later is high death rate of animals. Objective This experiment was to discuss the way which decrease the death of alloxan-injected animal and explore the effects of high blood glucose on the posterior capsular opacification (PCO). Methods Forty clean healthy male New Zealand white rabbits were randomly divided into 2 groups. 90mg/kg of alloxan were injected via ear vein once in 20 rabbits to create the diabetic animal models,and the equivalent amount of normal saline solution was injected at the same way as normal blood glucose group. The successful models were selected in the animals with the blood glucose level over 12. 0 mmol/L two weeks later, and PCO of lens were graded based on the method of Odrieh under the slit lamp. Extracapsular lens extraction was then performed on the right eye of rabbits in both groups, and the posterior capsules were obtained from these eyes at the 6th, 10th and 14th days after operation. The expression of proliferating cell nuclear antigen ( PCNA ) in posterior capsular lens epithelial cell was detected by immunohistochemistry. Results The modeling successful rate was 70% after injection of alloxan. The body weight of rabbits in high blood glucose group was significantly lowed and the blood glucose was significantly elevated in comparison with normal blood glucose group ( all P<0. 05). Two weeks after surgery ,2 eyes occurred 2 grade of PCO and only one eye showed the 1 grade of PCO in the high blood glucose group. However, 1 grade of PCO was found in 3 eyes in the normal blood glucose group. Biopsy revealed that PCNA was positively expressed in the cell nuclei of LECs in high blood glucose group rather than the normal blood glucose group from the 10th day after surgery. The proliferation index of PCNA was 0. 86±0. 04 and 0. 25±0. 03 respectively in high blood glucose group and normal blood glucose group, showing a significant difference between them (t = -16. 171 ,P = 0. 000). Conclusion Stable diabetic models of rabbits can be created by intravenous injection of 90 mg/kg alloxan. High blood glucose level is one of the important factors for the development of PCO.

OBJECTIVETo evaluate pathologically the effect of the single or combined application of UDP-glucose, GDNF and memantine on the improvement of white matter injury in neonatal rats with periventricular leukomalacia (PVL) under light and electron microscopy.

METHODSA five-day-old neonatal rat model for PVL was established by ligation of the lateral common carotid artery following 120-minute hypoxia. Rats were randomly divided into six groups (30 rats in each group): sham-operated, PVL, UDP-glucose (UDP-glucose 2000 mg/kg intraperitoneally after PVL), GDNF (GDNF 100 μg/kg intracerebrally after PVL), tmemantine (memantine 20 mg/kg intraperitoneally after PVL), and a combination administration of three drugs (UDP-glucose, GDNF and memantine). The rats were sacrificed 7 or 21 days after PVL for assessment of pathological changes in the white matter under both light and electron microscopy. The number and thickness of the myelin sheath in the white matter were measured under electron microscopy, and both of pathological grading and scoring were undertaken under light microscopy.

RESULTSThere was rare and sparse myelinogenesis with a loose arrangement of nerve fibers in the white matter under electron microscopy in the PVL group at 7 and 21 days after PVL. The number and thickness of the myelin sheath in the PVL group were significantly less than in the sham-operated, UDP-glucose, GDNF, memantine and combination administration groups (P<0.01). The results of pathological grading of white matter under light microscopy showed that all rats in the PVL group manifested either mild injury (38%-50%) or severe injury (50%-62%) at 7 and 21 days after PVL. The majority of rats (50%-88%) in the four drug administration groups had normal white matter at 7 and 21 days after PVL. The pathological scores at 7 and 21 days after PVL in the PVL group were the highest, and they were significantly higher than in the other five groups (P<0.05).

CONCLUSIONSThe single or combined application of UDP-glucose, GDNF and memantine may significantly improve pathological changes in the white matter of rats with PVL. The favorable effect is inferred to be closely correlated with the improvement of brain microenvironment and the enhancement of nerve regeneration promoted by the three drugs.

Animals ; Brain Ischemia ; drug therapy ; pathology ; Cerebral Ventricles ; pathology ; ultrastructure ; Female ; Glial Cell Line-Derived Neurotrophic Factor ; administration & dosage ; therapeutic use ; Humans ; Infant, Newborn ; Leukomalacia, Periventricular ; drug therapy ; Male ; Memantine ; administration & dosage ; therapeutic use ; Microscopy, Electron ; Rats ; Rats, Sprague-Dawley ; Uridine Diphosphate Glucose ; administration & dosage ; therapeutic use

OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

Abstract: Objective To explore and analyze the diagnostic value of multicolor melting curve analysis (MMCA) for the resistance of five anti-tuberculosis drugs, so as to clarify the clinical value of MMCA in detecting drug resistance of Mycobacterium tuberculosis. Methods From April 2021 to May 2022, 200 patients with positive Mycobacterium tuberculosis admitted to the Fourth People's Hospital of Qinghai Province were selected as research objects, and sputum specimens were taken from the patients. Traditional Mycobacterium tuberculosis drug sensitivity test (modified Löwenstein-Jensen medium method) and MMCA analysis were respectively given to detect the resistance of five anti-tuberculosis drugs, including isoniazid, ethambutol, streptomycin, rifampicin and isoniazid, respectively. Those samples with inconsistent results between the two diagnosis methods were subjected to gene sequencing verification, and the diagnosis efficiency of MMCA for the five anti-tuberculosis drugs was compared. Results Using Mycobacterium tuberculosis drug sensitivity as the gold standard for drug resistance diagnosis, the sensitivity of MMCA for detecting drug resistance of rifampicin, ethambutol, streptomycin, isoniazid and levofloxacin were 95.83% (46/48), 93.75% (15/16), 100.00% (15/15), 100.00% (20/20) and 70.00% (7/10), respectively, with statistical differences between groups (P<0.05). There were no statistically significant differences in the specificity, positive predictive value, negative predictive value and accuracy of MMCA for the five anti-tuberculosis drugs (P>0.05). For the 8 samples with inconsistent results between MMCA and modified Löwenstein-Jensen medium method, gene sequencing was performed and compared with the results of gene sequencing. After comparison with gene sequencing results, it was found that the coincidence rate of MMCA and gene sequencing results was 75.00% (6/8). Conclusions In the detection of drug-resistant mutations in TB patients, multi-color probe fusion curve analysis has high diagnostic efficacy for first-line anti-tuberculosis drugs, but is not sensitive to second-line anti-tuberculosis drug levofloxacin. Therefore, for the detection of first-line anti-tuberculosis drugs, MMCA has a good clinical application prospect.

BackgroundThe proliferation of lens epithelial cellsLECs) following extracapsular cataract extraction is the biological basis of posterior capsular collagen and cataract formation.Disintegrin is certified to competitively bind the integrin with extracellular matrix and therefore prevent the posterior capsular opacification (PCO).But,its molecular mechanism is below understand.ObjectiveThe present study was to investigative the effects of disintegrin (kistrin) on the expression of collagen in lens posterior capsular.MethodsThe right eyes of 24 New Zealand white rabbits received transparent lens extracapsular enudeation and were randomly divided into two groups using random number table,0.2 ml of kistrin ( 80 mg/L) was intracapsularly injected at end of the operation in 12 eyes ( kistrin group) and the same volume of normal saline solution was used at the same way in other 12 operative eyes ( normal saline group).The PCO was graded in postoperative 1,3,5,7,14 days on Odrich' s criteria under the slit lamp.The lens section was prepared at 14 days and 3 months after operation.Haematoxylin and eosin stain was used to examine the proliferation of LECs in posterior capsule; Masson stain was used to observe the collagenous fiber formation in capsule bag,and the expression of collagen type Ⅳ was detected by immunochemistry.Results No significant difference in the number PCO eye was found in postoperative 14 days between normal saline group and kistrin group ( P =0.093 ).However,the eye numbers of 2-3 grades of PCO were significant increased in normal saline group compared with kistrin group in 1,2,3 months after operation( P=0.041,0.014,0.022).In the operative 14 days,staining and adhesion of LECs in posterior capsule were more in normal saline group than kistrin group,and the fibrocytes in capsule were evidently increased in normal saline group in 3 months.Masson stain certified that the blue stain was seen to be stronger and more in posterior capsule in normal saline group in comparison with kistrin group in 3 months after operation,and the immunochemistry showed that the gray values of collagen type Ⅳ in posterior capsule were significant lower in normal saline group compared with kistrin group in both 14 days and 3 months after operation (P=0.000,0.001 ).ConclusionsKistrin can suppresses the proliferation of LECs and collagen type Ⅳ on rabbit lens posterior capsular after transparent lens extracapsular enudeation.

OBJECTIVETo explore whether the inhibitory effect of Genipin on uncoupling protein-2 (UCP-2) in mitochondria is involved in energy metabolism of androgen-independent PC3 prostate cancer cells.

METHODSPC3 prostate cancer cells were cultured and treated with Genipin at the concentrations of 40, 80, and 160 μmol/L for 48 hours. Then the proliferation of the cells was detected by MTT assay, the expression of UCP-2 mRNA determined by RT-PCR, and the content of intracellular pyruvic acid (PA) and the activity of succinate dehydrogenase (SDH) in the mitochondria measured by visible spectrophotometry.

RESULTSWith the increased concentration of Genipin, the proliferative activity of the PC-3 cells, the expression level of UCP-2 mRNA, the content of intracellular PA and the activity of SDH in the cells were all decreased, namely, with the enhanced inhibitory effect of Genipin on UCP-2, a trend of reduction was observed in the proliferation of the cells, intracellular PA content, and SDH activity in the mitochondria.

CONCLUSIONGenipin is involved in the energy metabolism of androgen-independent PC3 prostate cancer cells by reducing the content of intracellular PA and the activity of SDH in the mitochondria, which may be associated with its inhibitory effect on UCP-2.

Cell Line, Tumor ; drug effects ; Energy Metabolism ; Humans ; Ion Channels ; metabolism ; Iridoids ; pharmacology ; Male ; Mitochondria ; metabolism ; Mitochondrial Proteins ; metabolism ; Prostatic Neoplasms ; metabolism ; Pyruvic Acid ; metabolism ; RNA, Messenger ; Succinate Dehydrogenase ; metabolism ; Uncoupling Protein 2

OBJECTIVETo study in vivo the endogenous self-repair mechanism in immature white matter induced by ischemia in neonatal rats with periventricular leukomalacia (PVL).

METHODSFive-day-old neonatal Sprague-Dawley (SD) rats were randomly divided into sham and PVL groups. Rat model of PVL was prepared by ligation of the right common carotid artery following 2 hours of exposure to 8% oxygen. Pathological changes and myelination in the white matter were assessed under light and electron microscopy at 7 and 21 days after PVL. O4-positive OL precursor cells in the white matter were determined with immunofluorescence staining. Activation, proliferation, migration and differentiation of glial progenitor cells in SVZ were observed using immunofluorescent double labeling of either NG2 (marker of progenitor cells) and 5-bromodeoxyuridine (BrdU), or O4 (marker of OL precursor cells) and BrdU.

RESULTSAll rats in the PVL group manifested either mild or severe white matter injury under light microscopy, and had higher pathological scores of white matter compared with the sham group at 7 and 21 days after PVL (P<0.05). Electron microscopy showed that the number and thickness of myelin sheath in the PVL group were significantly reduced compared with the sham group (P<0.01). O4-positive OL precursor cells in the white matter observed under fluorescence microscopy were significantly reduced in the PVL group compared with the sham group (P<0.05). BrdU/NG2-positive cells in the SVZ increased significantly in the PVL group 48 hours after PVL and migrated into the periventricular area, reaching a peak on day 7 after PVL. BrdU/O4-positive newborn cells began to appear in the periventricular area 72 hours after PVL, and the number of BrdU/O4-positive cells in the PVL group was statistically more than in the sham group on day 21 after PVL (P<0.05).

CONCLUSIONSIschemia may induce brain self-repair in neonatal rats, resulting in activation and proliferation of NG2 glial progenitor cells in the SVZ migration and differentiation into OL precursor cells in periventricular white matter.

Animals ; Animals, Newborn ; Brain ; pathology ; Brain Ischemia ; pathology ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Disease Models, Animal ; Humans ; Infant, Newborn ; Leukomalacia, Periventricular ; pathology ; Myelin Sheath ; physiology ; Neuroglia ; pathology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; pathology

This study was purposed to investigate the immunoregulatory effect of endothelial cells derived from mesenchymal stem cells (MSC). The human MSC was induced to differentiate into endothelial cells for one week. The phenotypes were evaluated by flow cytometry, the cell morphologic feature was observed by invert phase-contrast microscope and analysis of capillary formation was performed by using the in vitro angiogenesis kit. The immunoregulatory effect was detected by lymphocyte transformation test. The result indicated that during the differentiation cells grew fast and there was no significant change in the phenotypes, i.e. CD73, CD105, HLA-ABC were positive and CD34, CD80, CD86, HLA-DR, CD31 were negative. Immunofluorescence analysis showed typical expression of the von Willebrand factor. Differentiated MSCs formed capillary-like structure. Endothelial cells derived from MSC also revealed immunosuppressive effect on T cell proliferation in a dose-dependent manner. It is concluded that endothelial cells derived from MSC also harbor immunoregulatory effect on T lymphocytes.
5'-Nucleotidase ; metabolism ; Cell Differentiation ; physiology ; Cells, Cultured ; Child ; Endothelial Cells ; cytology ; immunology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; T-Lymphocytes ; immunology ; von Willebrand Factor ; metabolism

OBJECTIVETo investigate the features of circulating plasmacytoid dendritic cells (pDCs) in patients with chronic hepatitis B for a better understanding of the immunopathogenesis of HBV infection.

METHODSFresh blood samples were collected from 20 patients with chronic hepatitis B (CHB) and from 15 healthy individuals who served as controls. pDCs were isolated from peripheral blood mononuclear cells (PBMCs) using immunomagnetic assay and detected by flow cytometry. Fresh PBMCs and isolated pDCs were stimulated in vitro using CpG ODN2216. The supernatants were measured for IFNa production using ELISA.

RESULTSThe peripheral pDCs frequency in CHB patients (0.192%+/-0.110%) was markedly lower than that in the healthy controls (0.287%+/-0.142%). After being pulsed with CpG ODN2216, the isolated pDCs produced lower levels of IFNa and expressed lower levels of CD80 and CD40 in the CHB patients when compared to those of the healthy controls. The level of IFNa was (972.6+/-705.5) pg/ml in the patients and (3 142.9+/-1 292.2) pg/ml in the controls. Moreover, the pDCs frequency was reversely correlated with serum ALT levels in these HBV infected patients.

CONCLUSIONThe reduced number and impaired function of circulating pDCs in patients with CHB may be related to their disease progression.

Adult ; Case-Control Studies ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; Female ; Flow Cytometry ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Interferon-alpha ; biosynthesis ; Male ; Middle Aged ; Oligodeoxyribonucleotides ; pharmacology