Objective To explore the function of fluorescent probe JC-1 in detecting the changes of mitochondrial membrane potential(△?m)in early apoptotic cells.Methods After 2-ME was used to induce MUTZ-1 cell apoptosis,cells were dyed with fluorescent probe JC-1,and then the changes of △?m in the early stage of apoptotic cells were analyzed by flow cytometry or detected under fluorescent microscope. Results The control cells with high △?m are those forming JC-1 aggregates in the inner membrane of mitochondria,thus showing orange-red fluorescence.2-ME caused decrease of △?m in MUTZ-1 cells,in which JC-1 maintains monomeric form,thus showing only green fluorescence.The decreases of △?m were in a time-dependent manner,which were significantly higher than those in control group(P

In order to further improve the curative effect of the herb-participated moxibustion on umbilicus, we collected a wide range of literature on herb-participated moxibustion on umbilicus and then systematically arranged them to analyze and summarize the technology and operating methods of herb-participated moxibustion on umbilicus. We also briefly discussed issues on application of medicine, locust tree skin and ginger, the method of kneading dough for medical uses, and the appropriate size of moxa cone and its using frequency in order to form clear concepts and standardized operations to provide theories and operational basis for the clinical application of modern herb-participated moxibustion on umbilicus.
China ; History, Ancient ; Humans ; Medicine in Literature ; Medicine, Chinese Traditional ; history ; Moxibustion ; history ; instrumentation ; methods ; Plants, Medicinal ; chemistry ; Umbilicus ; physiopathology

Objective To assess the efficacy of one-stage posterior median incision via costotransverse joint in treating thoracic spinal tuberculosis.Methods Thirty three patients with tuberculosis of thoracic spine undergoing one-stage posterior thoracic spine debridement,bone grafting fusion and posterior instrumentation from July 2011 to October 2013 were included in the study.There were 18 males and 15 females.The age was from 17 to 39 years old with an average of 29.5.The course was from 5 to 11 months with an average of 7.3.The Cobb angle was from 19° to 42° with an average of 29.8°.There were 5 in upper thoracic spine,17 in middle thoracic spine and 11 in lower thoracic spine.6 were in Frankel scale grade C,11 were in grade D and 16 were in grade E before surgery.Postoperative kyphosis correction,recovery of neurological function and bone fusion were observed.Results All the surgeries were completed successfully.The operation time was from 120 to 230 min with an average of 183.The blood loss during operation was from 480 to 700 ml with an average of 530.All 33 patients were followed-up for 12-36 months with an average of 22.7 months.The Cobb angle was from 8° to 15° with an average of 10.2°,the correction rate was 77％.The Frankel scale of 11 patients recovered from D to E,2 recovered from C to D and 4 recovered from C to E.The postoperative kyphosis correction and Frankel scale were significantly improved,all patients had a 100％ bone fusion rate and there were no internal fixation loosened or shift of graft bone at the last follow-up.ESR and CRP were returned to the normal range and the tuberculosis symptoms disappeared.Conclusions One-stage posterior median incision via costotransverse joint can complete surgery by the same position and the same incision in treating thoracic spinal tuberculosis with safety and good clinical efficacy.

Objective To assess the clinical efficacy of a modified complete retroperitoneal laparoscopic nephroureterectomy via 3 port approach.Methods From August 2013 to February 2016,23 patients with complete retroperitoneal laparoscopic renal and ureteral sleeve resection were treated with modified three port approach,including 15 males and 8 females.The average age was 67 years old (ranging 44-83 years old).All patients had complained about the hematuria before operation and urine exfoliated cells showed moderate to severe nuclear atypia.All patients accepted the abdominal CT and urography CTU examination,pre-operatively.All of them was diagnosed localized upper urinary tract malignant tumors based on those images,including 13 cases in the pelvis,and 10 cases in the upper segment of the ureter.No chemotherapy,radiotherapy or immunotherapy was performed before surgery.No patients have the history of severe basic disease or upper urinary procedure.The operations were performed under general anesthesia,patients take the contralateral back 30 degrees slope,low elevation head foot,waist bridge,side waist stretch.In the anterior superior iliac spine perpendicular to the line 2 cm parallel to the lower intersection of the rib border were disposed into the 12 mm trocar.Above the anterior superior iliac spine two cross finger level with the intersection of the anterior axillary node,we placed into the 10 mm trocar placement lens.Laparoscopic placement of third casing form an isosceles triangle with the first two casing.The renal fascia was incised with an ultrasonic knife from the renal dorsal side,and the renal hilum was isolated from the kidney by suction aspirator.The renal artery and vein were separated and closed by hem-o-lok.Along the psoas muscle surface to ureter,ureteral clipping by hem-o-lok but not to cut off the free distal ureteral,the lens is composed of first casing into,using ultrasonic knife to free ureter to the bladder wall segment,with 30 mm endoscopic stapler ureter and bladder wall cut off part.Operation time,blood loss and postoperative recovery were recorded in 23 cases.Results All 23 cases were successfully operated without related the operative complication.The operative duration ranged from 3.5 to 6.1 h (mean 4.8 h),the blood loss was 30-880 ml (mean 304 ml),and the postoperative stay was 8-30 d (mean 17.8 d).There are 3 cases of positive lymph node by postoperative pathological reports.Within 2 to 30 months following up,2 patients died of tumor progression in 6 months after surgery.4 patients were diagnosed with bladder cancer in 15 months,15 months,21 months,24 months after surgery,respectively.And the transurethral resection of bladder tumor was performed.Conclusion The modified complete retroperitoneal laparoscopic nephroureterectomy via three ports is safe and reliable.

· AIM: To investigate the role of intercellular adhesion molecule-1 (ICAM-1) in the graft rejection of rabbit penetrating keratoplasty (PKP).· METHODS: Sutures were used to produce corneal neovascularization (CNV) on New Zealand white rabbits to make PKP model. Graft mean survival time and rejection index were determined. The levels of sICAM-1 in serum and aqueous humor were dynamically determined by enzyme-linked immunosorbent assay (ELISA).Immunohistochemical technique was used on cornea to get the evidence of expression and distribution of ICAM-1.· RESULTS: Graft mean survival time is (12.4±1.3) days.The mean sICAM-1 levels in aqueous humor and serum in the control animals that did not undergo surgery were (16.6±3.6)ng/L and (95.2±6.3) ng/L respectively. Levels in aqueous humor and serum for the graft rejection group increased postoperatively, continued to increase to (53.9±19.2)ng/Land (378.8±30.6)ng/L, compared to those in the other group (P ＜0.01). ICAM-1 positive cells were found in the graft rejection group.· CONCLUSION: ICAM-1 plays a critical role in initiating and maintaining corneal graft rejection. Sequential determination of serum sICAM-1 after operation may be of value in the prediction and diagnosis of acute rejection.

Antibodies, Viral ; blood ; Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Fever ; Humans ; Leukoencephalitis, Acute Hemorrhagic ; diagnosis ; therapy ; Male ; Prognosis

Objective High-throughout sequencing of all plasmid of 206 strains of Klebsiella pneu-moniae using Solexa/Illumina sequencing technology in order to investigate the resistance for plasmids in Klebsiella pneumoniae. Methods Bacterial isolates were obtained over the years 2002-2008. Solexa/Illumi-na sequencing technology was used to sequence both samples (S1 and S2) to a depth of between 10-560 fold coverage. We used SOAP provided by BGI to find SNPS and use velvet package to assemble these sequences and gained some long sequences, and MAQ programs developed in the laboratory were used to annotate SNPs and compare lineage-specific mutations in SHV-ESBLs. Results The Metagenome of plasmid encodes a 13 variety of resistance-related genes with exceptionally high copy numbers, including ABC-type efflux pumps and 4 variety of β-lactamases, SHV-ESBLs is abroad presence. We systematically investigated single nucleo-tide substitutions in plasmids metagenome, and found an amount of nonsynonymous mutations in the SHV-ESBLs genes. Conclusion Probabily in press of positive selection, we can clearly see these nonsynonymous changes predominantly occurred in plasmid SHV-ESBLs genes. And our findings indicate a unspecial low-level resistance contribute to antimicrobial efflux in the metagenome of plasmid in Klebsiella pneumoniae.

OBJECTIVETo sum up the clinical experience in the management of benign prostatic hyperplasia (BPH) with the prostate weighing over 80 ml by transurethral resection of the prostate (TURP) combined with 2 μm continuous-wave laser vaporesection (LVR).

METHODSWe retrospectively analyzed the clinical effects of TURP combined with 2 μm LVR in the treatment of 46 cases of BPH with the prostate volume > 80 ml.

RESULTSAll the operations were successfully accomplished. The operation time and intraoperative blood loss were (112.0 ± 20.0) min (range 86-176 min) and (77.9 ± 25.9) ml (range 50-200 ml), respectively. The catheters were withdrawn at 7 days after surgery. Transient urinary incontinence occurred in 6 cases and secondary hemorrhage was found in 2 postoperatively. Six-month follow-up revealed no urethral stricture or other complications. Compared with the baseline, the international prostate symptom score (IPSS) was significantly decreased at 6 months after operation (26.3 ± 1.8 vs 11.6 ± 1.7, P <0.05), and so were the quality of life (QOL) score (5.3 ± 0.7 vs 1.3 ± 1.1, P <0.05) and post-void residual urine (PVR) ([115.5 ± 55.6] ml vs [19.9 ± 11.6] ml, P <0.05). However, the maximum urinary flow rate (Qmax) was remarkably increased from (4.1 ± 2.6) ml/s to (16.2 ± 1.7) ml/s (P <0.05).

CONCLUSIONTURP combined with 2 μm LVR is safe and effective for the treatment of BPH with the prostate volume >80 ml.

Aged ; Blood Loss, Surgical ; Humans ; Laser Therapy ; methods ; Male ; Middle Aged ; Organ Size ; Prostate ; pathology ; Prostatic Hyperplasia ; pathology ; surgery ; Quality of Life ; Retrospective Studies ; Transurethral Resection of Prostate ; methods ; Treatment Outcome ; Urethral Stricture ; Urinary Incontinence ; etiology ; Urinary Retention

OBJECTIVETo study the effect of 2-methoxyestradiol(2-ME) on telomerase activity expression and apoptosis in human myelodysplastic syndrome cells line MUTZ-1.

METHODSMUTZ-1 cells were incubated with different concentrations of 2-ME, apoptosis rate and cell cycle were measured by flow cytometry (FCM). Telomerase activity in MUTZ-1 cells was examined by telomeric repeat amplification protocol-Enzyme linked immunosorbent assay (TRAP-ELISA).

RESULTSThe FCM analysis showed that cells in G0/G1 phase and S phase were decreased, while in G2/M phase increased after exposed to 1,2 and 4 micromol/L of 2-ME for 12 hours (P < 0.05). 1 and 2 micromol/L of 2-ME had no notable effect on MUTZ-1 cells as compared with the control group (P > 0. 05). Cells incubated with 1, 2 and 4 micromol/L of 2-ME for 36 hours were induced apoptosis, the percentage of apoptosis was between (12.87 +/- 0.86)% and (21.82 +/- 1.71)% with a dose- and time- dependent manner. Telomerase activity was significantly inhibited in these concentration and negatively correlated with cell number in G2/M phase (r = -0.979, P = 0.021) and increased apoptosis (r = -0.970, P = 0.030 ), respectively. Moreover, the inhibition effect of telomerase activity was enhanced in a dose- and time- dependent manner.

CONCLUSIONS2-ME-induced apoptosis and inhibition of telomerase activity provide a possible mechanism for explaining the 2-ME's anticancer activity.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; Myelodysplastic Syndromes ; enzymology ; pathology ; Telomerase ; metabolism

This study was aimed to investigate the reversible effect of tetrandrine, toremifene and their combination on multidrug resistance of K562/A02 cell line. The IC(50) (the concentration causing 50% inhibition of cell growth) of adriamycin (ADR) were assayed by MTT method, the expression of MDR1 mRNA was measured by RT-PCR, the concentration of p-glycoprotein (P-gp) and intracellular ADR were detected by flow cytometry. The results showed that the IC(50) of ADR on K562/A02 and K562 cells were 57.43 and 1.16 mg/L, respectively. The IC(50) of ADR on K562/A02 cells after treatment with tetrandrine, toremifene and both combination were 14.12, 20.74 and 9.14 mg/L respectively, but both drugs did not influence the IC(50) of ADR on K562 cells. Pretreating K562/A02 cells with toremifene (2.5 micromol/L), tetrandrine (1 micromol/L) or both for 72 hours partially restored the sensitivity of K562/A02 cells to ADR. Tetrandrine and toremifene (alone or combination) elevated the ADR concentration in K562/A02, down regulated the expressions of P-gp and MDR1 mRNA. It is concluded that multidrug resistance of K562/A02 cells can be partially reversed by tetrandrine or toremifene, the combination of both drugs shows a higher synergistic reversal effect.
Antineoplastic Agents, Hormonal ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Benzylisoquinolines ; pharmacology ; Doxorubicin ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; Toremifene ; pharmacology