AIM: To investigate the expression of matrix metalloproteinases (MMPs), membrane type MMPs (MT-MMPs) and their inhibitors (TIMPs) in human primary cultured prostatic cells and malignant prostate cell lines.METHODS: Reverse transcription-polymerase chain reaction-based measurements of the mRNA levels of MMP-2, MMP-7, MMP-9, MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 in relation to the house-keeping gene glyceraldehydes phosphate dehydrogenase were performed in cancerous and non-cancerous prostatic tissue samples, in primary cell cultures of epithelial cells, in both fibroblasts and smooth-muscle cells as stromal cells, and in the human malignant prostatic cell lines DU-145, LNCaP and PC-3.RESULTS: MMP-2 was mainly expressed in stromal cells. MMP-7 and MMP-9 showed their highest values in epithelial cells. MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 were found both in stromal and in epithelial cells, but there were some differences between the expression in fibroblasts and smooth-muscle cells. Different expression was also observed between the cells deriving from the primary cell cultures, the benign cell line BPH-1, and the malignant cell lines LNCaP, DU-145, and PC-3.CONCLUSION: These results concerning different expression of MMPs and TIMPs in cells from prostatic tissue suggest that a better insight into changes observed in prostatic tissue needs studies on cells cultured from the tissue.

OBJECTIVETo observe preliminary efficacy of decompressive unilateral improved transforaminal lumbar interbody fusion (TLIF) for the treatment of lumbar degenerative diseases.

METHODSFrom August 2009 to December 2011, 28 patients with lumbar degenerative diseases were treated by decompressive unilateral improved TLIF,including 16 males and 12 females with an average of 61 (aged 46 to 71) years old,the courses of disease ranged from 6 months to 6 years. Among them, 20 cases suffered from lumbar spinal stenosis, 8 cases were lumbar disc herniation. Decompressive range included single segment in 24 cases,and double segments in 4 cases; 15 cases were performed operation on the left side, 13 cases on the right side. JOA lower back pain scoring system (29 points) were applied for evaluate preoperative and postoperative symptoms, physical signs and sphincteral functions;Visual analogue scale (VAS) were used to evaluate preoperative and postoperative low back pain.

RESULTSAll patients were followed up 6 to 28 (mean 14) months. Postoperative JOA score and VAS score were 17.9 +/- 2.2, 2.8 +/- 0.7 respectively,and preoperative JOA score and VAS score were 8.5 +/- 1.7, 8.6 +/- 1.2, respectively. There were significant meaning in JOA and VAS scores before and after operation (P < 0.05). Twenty-eight patients were all obtained intervertebral synostosis.

CONCLUSIONDecompressive unilateral improved TLIF for treatment of unilateral radicular lumbar spinal stenosis and lumbar disc herniation,which has advantages of minimally invasive,curative effects,decrease medical costs,is worthy spreading in clinical.

Adult ; Aged ; Decompression, Surgical ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Diseases ; surgery ; Spinal Fusion ; Treatment Outcome

Objective:To investigate the compliance with early Warfarin anticoagulation therapy after cardiac mechanical valve replacement(MHVR)and its related factors in elderly patients.Methods:The convenience sampling method was used to prospectively recruit 210 patients undergone MHVR at the Ningbo Medical Center of Lihuili Hospital from January 2017 to October 2019.Six months after discharge, face-to-face interviews or telephone follow-ups were conducted to assess general information, warfarin anticoagulation knowledge, anticoagulant treatment compliance and social support.Results:The overall compliance of early Warfarin anticoagulation therapy was excellent, with 99.5% of patients compliant with medication and 99.0% compliant with INR monitoring, both higher than the rate of compliance with advised lifestyle adjustment(92.1%). Anticoagulation knowledge and age were the main influencing factors for compliance in elderly patients after MHVR.Conclusions:The compliance with early Warfarin anticoagulation therapy after MHVR is good in elderly patients in the Ningbo area.The correlation analysis suggests that medical professionals need to promote education on anticoagulation knowledge and pay more attention to anticoagulation compliance in elderly people.

The single needle method for urethrovesical anastomosis with strengthened posterior fixation during laparoscopic radical prostatectomy was explored. The method was initiated by performing a fixing suture with a knot at 4 o'clock of the posterior lip of bladder neck, and another suture at nearby position was performed to leave the knot outside. From 5 o'clock to 8 o'clock, sutures were performed every one o'clock to secure posterior approximation, then every two o'clock a suture. To avoid a loose anastomosis, lock sutures were performed every 3 sutures. The needle was always driven full-thickness outside-in in the bladder neck and inside-out on the urethra. After completing the full circumference, the needle was drawn near the 4 o'clock and tied at the tail end. Any leakage could be closed with additional interrupted sutures. The clinical data of 89 patients who underwent this method were retrospectively compared with those of 23 patients who underwent the single knot method. The results showed that the anastomosis, operative and catheterization time was 17.6+/-4.7 min, 134.0+/-10.7 min and 6.5+1.6 days respectively. There were 3 temporal urinary leakages identified in 89 cases requiring prolonged catheterization. No urinary leak and anastomotic stricture was confirmed, and 95.2% patients had total urinary control. It was concluded that this method was simple and safe for urethrovesical anastomosis.

Objective To describe the single needle running suture method for the urethrovesi-cal anastomosis during laparoscopic radical prostatectomy(LRP). Methods Forty-five patients of prostate cancer underwent LRP with the single needle running suture method. The technique was initi-ated by performing a fixing suture at the posterior lip of bladder neck at 4 o' clock and tying the first knot. Another suture at the nearby position of the first suture was performed to leave the first knot outside. From 5 o' clock to 8 o' clock, sutures were performed every one o' clock to secure posterior approximation, then every two o'clock a suture. To avoid a loose anastomosis, lock sutures were per-formed every 3 sutures. After completing the full circumference, the needle was drawn at the 2 o' clock for the second knot. The needle was always driven full-thickness outside-in in the bladder neck and inside-out on the urethra. Any remaining leakage could be closed with additional interrupted su-tures. Results All urethrovesical anastomosis were completed successfully. The mean anastomosis time was 16 rain(from 12 to 25 min), and mean operative time was 132 rain (112 to 185 rain). The mean catheterization time was 9 d(7 to 14 d). Three temporal urinary leaks requiring prolonged cathe-terization were identified. Forty-four patients had total urinary control in 1 year postoperatively and no other short-term or persistent complication was found with a mean follow-up of 21 months. Conclu- sion The single needle running suture method could be a simple and safe method for urethrovesical anastomosis during LRP.

Objective To explore the expression and diagnostic value of serum microRNA-574-3p (miR-574-3p) in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC).Methods A total of 90 patients with HBV-related HCC,40 patients with HBV-related liver cirrhosis (LC) and 50 healthy controls were recruited.The expression levels of serum miR-574-3p,α-fetoprotein (AFP) and α-Lfucosidase (AFU) of all subjects were determined.The difference of serum miR-574-3p level between groups was compared.The relation between serum miR-574-3p level of HCC patients and its clinical pathological characteristics was analyzed.The t-test was performed.The relationship between serum miR-574-3p level of HCC patients and AFP,AFU was analyzed by Spearman correlation analysis.The diagnostic efficacy of them as diagnostic markers was evaluated by receiver operating characteristic curves (ROC) and the area under the curve (AUC)(95％ CI (confidence interval)).Results The relative quantity expression of serum miR-574-3p of HCC group,LC group and healthy group was 2.152(1.654,3.061),1.292 (0.984,1.666) and 1.018 (0.750,1.726),respectively.The expression level of serum miR-574-3p of HCC group was significantly higher than those of LC group and healthy control group,and the differences were statistically significant (t=2.726 and 2.845,both P＜0.01).The expression level of serum miR-574-3p of HCC patients was significantly different in tumors with different degree of differentiation (t=2.262,P=0.039),different stage (t=2.354,P=0.025) and different HBV DNA concentrations (t=2.771,P＜0.01).There was no correlation between serum miR-574-3p level and AFP (r2 =0.076,P=0.505),AFU (r2 =0.082,P=0.422) in HCC patients.When compared HCC group with LC group,AUC of serum miR-574-3p of was 0.823 and 95％ CI was 0.750 to 0.897.When compared HCC group with healthy control group,AUC of serum miR-574-3p was 0.840 and 95％CI was 0.769 to 0.910.Conclusions The expression level of serum miR-574-3p of HCC patients is significantly higher than those of LC patients and healthy controls.Serum miR-574-3p may be an important biomarker in the diagnosis of HCC.

The expression of KAI1/CD82 and MRP-1/CD9 in transitional cell carcinoma of bladder (TCCB) and its clinical significance were investigated. Immunohistochemistry was used to detect KAI1/CD82 and MRP-1/CD9 protein expression in 52 TCCB specimens. Correlation between the expression of KAI1/CD82 and MRP-1/CD9 to clinicopathologic factors was statistically analyzed. The results showed that the positive rate of KAI1/CD82 and MRP-1/CD9 in TCCB was 50% and 61.5%, respectively. The MRP-1/CD9 and KAI1/CD82 expression was significantly associated with grade of TCCB (P<0.05), but no correlation was found between MRP-1/CD9 or KAI1/CD82 expression and clinical stage of TCCB (P>0.05). The expression level of MRP-1/CD9 and KAI1/CD82 in recurrent TCCB samples was lower than that in non-recurrent samples (P<0.05). Meanwhile, the correlation between the KAI1/CD82 expression and MRP-1/CD9 expression was statistically significant (r=0.316, P<0.05). It was concluded that KAI1/CD82 and MRP-1/CD9 expression may be important prognostic indicators and potentially useful for assessing the biological behavior of TCCB.

Objective To investigate the genetic polymorphisms of 3 short tandem repeat (STR) loci D18S53, D18S59 and D18S488 on chromosome 18 in fetus of Tianjin Han population, and to provide basic data in the use of 3 STR lo-ci in the prenatal diagnosis of Edward syndrome (ES). Methods A total of 64 villus samples and 374 amniotic fluid sam-ples were collected from gravida in Tianjin Han population. QF-PCR and ABI PRISM 377 sequence were used in this study. The frequencies of the genotypes were tested with H-W equilibrium. Genetic analysis was performed to conclude some data of population genetics such as the frequency of the alleles, the heterozygosity of observation (Ho), the polymorphism informa-tion content (PIC), the probability of discrimination power (DP), and the probability of exclusion (PE). Results The 15, 13 and 15 alleles of D18S53, D18S59 and D18S488 were observed respectively. The frequencies of the genotypes were in good agreement with H-W equilibrium. The Ho of 3 STR loci were 0.797, 0.847 and 0.792. The PIC was 0.81, 0.75 and 0.73. The DP was 0.944, 0.901 and 0.881. The PE was 0.593, 0.689 and 0.585. Conclusion D18S53, D18S59 and D18S488 STR lo-ci were the favorable genetic markers of chromosome 18, which can be used in prenatal genetic diagnosis of ES.

Objective To study the effects on berberine hydrochloride on the mitochondria membrane potential and free intracellular calcium of HaCaT cells, and elucidate the mechanism of action of berberine on keratinocytes. Methods Rhodamine-123 fluorescence (very sensitive to mitochondria membrane potential) and Fluo-3/AM fluorescence (suitabe to detect free intracellular calcium in single HaCaT cell) were measured by laser scanning confocal technique. Results Fluo-3/AM fluorescence intensity of HaCaT cells was persistently increased after treating with berberine at concentrations of 5 ? 10-5M, 2.5 ? 10-5M and 1.25 ? 10-5M, and significant differences were observed as compared with the PBS control. The intensity of rhodamine-123 fluorescence in HaCaT cells was decreased immediately when exposed to berberine, with significant difference from that of the PBS control. Conclusions It is suggested that berberine could increase free intracellular calcium and decrease mitochondria membrane potential of HaCaT cells, induce overload of intracellular calcium, influence energy metabolism, and then inhibit the proliferation of keratinocytes.

AIM:To characterize the effect of estradiol on proliferation,differentiation and extracellular matrix(ECM) accumulation in stromal cells through regulation of BPH-1 paracrine.METHODS:BPH-1 cells were stimulated with different concentrations of estradiol.Conditioned media(CM) were harvested and their effects on stromal cell cultures were tested.Cell proliferation was determined by MTT assay.mRNA of smoothelin,fibronectin,collagen Ⅳ and transforming growth factor ?1(TGF-?1) were analyzed by real-time RT-PCR.Western blotting was used to determine smooth muscle myosin heavy chain(SMMHC).ELISA and radioimmunoassay were respectively used to measure fibronectin,TGF-?1 and collagen Ⅳ protein expressions.RESULTS:Estrodiol stimulated the expression and secretion of TGF-?1 in BPH-1 cells.The proliferation of stromal cells increased when they were cultured with CM harvested from estrogen treated BPH-1 cells.The mRNA levels of collagen Ⅳ and smoothelin increased in stromal cells treated with CM from BPH-1 cells.The results of radioimmunoassay also showed that the collagen Ⅳ protein level up-regulated in the supernatants and cell extracts of CM-treated stromal cells.A neutralizing antibody to TGF-?1 inhibited the stimulation of collagen Ⅳ and SMMHC by BPH-1 CM.The expression of fibronectin was only marginally changed in stromal cells cultured in the presence of BPH-1 CM.CONCLUSION:The BPH-1 cells increase ECM accumulation and differentiation of stromal cells through TGF-?1.Estradiol stimulate differentiation of stromal cells by induction of TGF-?1 expression.Estradiol stimulate proliferation by influencing the factors secreted from prostatic epithelial cells.