Objective To study the effect of methotrexate on the neointimal formation after arterial balloon injury. Methods Male rabbits were randomized into four groups: MTX 0.5 mg/kg per week, MTX 1.5 mg/kg per week, MTX 5 mg/kg per week and the control. Drugs were administered by intramuscular injection. Rabbit carotid arteries were harvested 2 and 4 weeks after injury. Results Histologically, the intimal areas were reduced significantly in MTX treated animals compared with the controls. The VSMC proliferation in injured vessels was identified by immunostaining for proliferating cell nuclear antigen (PCNA). In comparison, PCNA-positive cells in both intima and media were significantly reduced by treatment of MTX. But MTX did not enhance reendothelialization in the injured carotid arteries as determined by Evans blue stain.Conclusion Low dosage of MTX could attenuate neointimal formation after arterial injury by inhibiting VSMC proliferation.

Autophagy is an important homeostatic cellular recycling mechanism responsible for degrading injured or dysfunctional cellular organelles and proteins in all living cells. Aging is a universal phenomenon characterized by progressive deterioration of cells and organs due to accumulation of macromolecular and organelle damage. Growing evidences indicate that the rate of autophagosome formation and maturation and the efficiency of autophagosome/lysosome fusion decline with age. Dysfunctional autophagy has also been observed in age-related diseases. Autophagy disruption resulted accumulation of mutated or misfolded proteins is the essential feature of neurodegenerative disorders. However, in cancers, fibroproliferative diseases or cardiovascular diseases, autophagy can play either a protective or destructive role in different types of disease, and even in different stages of the same disease. The review will discuss the cellular and molecular mechanisms of autophagy and its important role in the pathogenesis of aging and age-related diseases, and the ongoing drug discovery strategies for therapeutic intervention.
Aging ; Autophagy ; Drug Discovery ; Humans ; Lysosomes ; metabolism ; Neurodegenerative Diseases ; Phagosomes ; metabolism ; Protein Folding

Objective To compare the perioperative outcomes and short-term efficacy of three-port extraperitoneal laparoscopic radical prostatectomy (ELRP) and four-port ELRP.Methods Two hundred patients who had undergone ELRP for prostate cancer by a single surgeon from November 2010 to October 2014 were retrospectively analyzed.Among them,95 cases underwent three-port ELRP and 105 cases underwent four-port ELRP.On the basis of traditional four-port ELRP,three-port ELRP was characterized by the omission of the trocar on the inner side of right anterior superior iliac spine.The mean age was 66.8 ± 15.5 years,and mean total prostate specific antigen (tPSA) was 15.3 ± 12.4 μg/L.There were no significant differences including age,body mass index,tPSA,clinical stages,acceptance of neoadjuvant hormone therapy,history of transurethral resection of the prostate,history of diabetes mellitus between the 2 groups (P ＞ 0.05).Patients in three-port ELRP group had significantly smaller prostate volume than fourport group (35.6 ± 16.7 ml versus 42.2 ± 24.7 ml,P ＜ 0.05).The clinical factors as operative time,estimated blood loss,hospital stay,drainage tube keeping days,pathological Gleason scores,pathological stages,positive surgical margin rates,biochemical recurrence rates and urinary incontinence rates were compared between the 2 groups.Results The three-port group had significantly shorter operative time than the four-port group (81.0 ± 18.6 min versus 103.6 ±34.6 min),less estimated blood loss (102.6 ±75.8 ml versus 217.5 ± 182.9 ml),less positive surgical margin rates (13.7％ versus 27.6％).There were 9 patients having Gleason scores more than 7 in the three-port ELRP group and 29 patients in four-port ELRP group (P ＜ 0.05).There were no significant differences of hospital stay,drainage tube keeping days,pathological stages between the 2 groups (P ＞ 0.05).Eighty-three cases in the three-port ELRP group (87.4％) were followed up for 5-19 months with the median time of 11 months.Ninety-two cases in fourport ELRP group (87.6％) were followed up for 17-52 months and the median time was 27 months.There were no significant differences of biochemical recurrence rates and urinary incontinence rates between the 2 groups(P ＞ 0.05).Conclusions Compared to four-port ELRP,three-port ELRP can provide shorter operative time,less blood loss,better negative surgical margin rates,similar oncological control and recovery of postoperative continence.In experienced hands,three-port ELRP could be a feasible and effective option for localized prostate cancer.

Objective To evaluate the learning curve of three-port extraperitoneal laparoscopic radical prostatectomy(ELRP) and to minimize operative time and blood loss about this procedure.Methods From August 2013 to October 2014,the data from 95 consecutive patients,who had undergone three-port ELRP for prostate cancer,were retrospectively analyzed.The mean age was 65.9 ± 7.7 years,mean total PSA level was 15.4 ± 12.7 μg/L,and mean body mass index(BMI) was 24.8 ± 3.2 kg/m2.According to the number of procedures performed by the surgeon,all patients were classified into three chronologic groups,including group A (No.1-32),group B (No.33-64) and group C (No.65-95).There were no significant differences including age,BMI,tPSA,estimated prostate volume,clinical stages,history of neoadjuvant endocrine therapy,history of transurethral resection of the prostate (TURP) among group A,B and C (P ＞ 0.05).The operative outcomes analyzed were operative time,estimated blood loss,hospital stay,drainage tube indwelling days,pathological Gleason scores,pathological stages,positive surgical margin rates,biochemical recurrence rates and urinary incontinence rates.Among these 95 patients,the results of the first 32 cases were compared with those of the remaining 63 cases,the first 64 with the remaining 31.Results The average operative time in 95 patients was 81.0 ± 18.6 min.The sloping learning curve for this surgeon showed that the operative time for all 95 cases was strongly correlated with additional experience (| rs | =0.612,P＜0.01).Operative time,however,was not strongly correlated with the surgeon's experience in each group of A,B and C(P ＞0.05).Group A had longer operative time than that of Group B plus C(96.4 ± 11.3 min vs 73.2 ± 16.7 min,P ＜0.01).Group A plus B had longer operative time than that of group C (87.6 ± 17.2 min vs 67.5 ± 13.8 min,P ＜ 0.01).For all cases,the estimated blood loss was strongly correlated with additional experience (| rs | =0.677,P ＜ 0.01).Estimated blood loss was strongly correlated with the accumulation of experience for the initial 32 cases(| rs | =0.619,P ＜ 0.01).However,no strong correlation was observed over the next 63 cases.Group A had more blood loss than that of Group B plus C (158.7 ± 81.3 ml vs 74.1 ± 54.4 ml,P ＜ 0.01).Group A plus B had more blood loss than that of group C (125.5 ± 71.6 ml vs 55.3 ± 61.6 ml,P ＜ 0.01).But hospital stay,drainage tube keeping days were not strongly correlated with additional experience in each group(P ＞ 0.05).There were no significant correlation between the accumulation of experience and positive surgical margin rates,biochemical recurrence rates and urinary incontinence rates.Conclusion Our experience of three-port ELRP cases appears to be favorable with decreasing tendency in operative time,estimated blood loss with experience accumulation.Exposure to 32 surgeries,operative time and estimated blood loss reduced significantly,and after 64 cases operative time and estimated blood loss further reduced.

Objective To construct and screen effective shRNA expression vectors targeting human AQP1 gene ,and evaluate the interference efficiency of the AQP1 shRNA recombinant plasmids ,thus provide basis for further exploration on the effect and mechanism of AQP1 gene on human breast cancer cells .Methods Four pairs of shRNA sequences targeting human AQP1 gene were designed and synthesized ,and then inserted into the GV115 vector .AQP1 shRNA and control shRNA plasmids were trans‐fected into human breast cancer MCF‐7 cells .The expression of AQP1 mRNA and protein were detected by real time PCR(RT‐PCR) and Western blot to evaluate the interfering efficiency .Results RT‐PCR demonstrated that AQP1 was expressed in human breast cancer MCF‐7 cells .Sequencing showed that the shRNA vectors targeting AQP1 were successfully constructed .48 h after the AQP1 shRNA transfection ,AQP1 mRNA and protein expression levels in MCF‐7 cells were reduced to a significant degree ,and the AQP1 shRNA 4 plasmid vector could inhibit the AQP1 most efficiently .Conclusion The AQP1 shRNA recombinant plasmids vectors were successfully constructed and can significantly inhibit the expression of AQP1 in MCF‐7 human breast cancer cells .

Lactic acid production of twelve strains of LAB isolated from broiler intestine and tolerance property of three strains were investigated. The results of lactic acid production showed that among all strains K6 exhibited the most rapid production during the first twelve hours, the seconds were K9 and C1; D17 exhibited the highest production of lactic acid by twenty-four hours, C1 exhibited the highest production of lactic acid by forty-eight hours. The pH values in three strains of K9、D17 and C1 culture showed the fast decline during the first twelve hours, with the final values significantly lower than those of other strains cultures. The results of tolerance property showed that the survival counts of C1could be detected when pH value was at 2 after three hours, but the survival counts of D17 and K9 could not be detected after one hour. When pH value was at 2.5 after three hours ,the survival counts of C1 declined from 10~ 8.2 /mL to 10~ 4.8 /mL, K9 from 10~ 8.2 /mL to 10~ 4.6 /mL, the survival counts of D17 could not be detected. 0.08% bile had few effects on the survival counts of three strains; when incubated in the medium with 0.40% bile, the survival counts of C1 declined from 10~ 8.4 /mL to 10~ 6.5 /mL,D17 from 10~ 10.3 /mL to 10~ 7.5 /mL, and K9 from 10~ 9.8 /mL to 10~ 7.7 /mL. When the group treated with 37℃ for 20 minutes was served as the control, the survival counts of C1 and K9 was not detected when treated with 80℃, but the survival counts of D17 were 10~ 4.9 /mL, when treatment with 65℃ the survival counts of C1 and K9 decreased significantly .

The nuclear transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) plays a crucial role in maintaining cellular redox homeostasis. The aberrant NRF2 signaling confers enhanced antioxidant capacity, which is linked to tumor progression and therapeutic resistance. The current study investigates the biological effects and molecular mechanism of tribbles homolog 3 (TRIB3), a stress-induced protein, in regulating cell survival and apoptosis in lung cancer. This study first performed the RNA sequencing data analysis with 576 lung adenocarcinoma patients from the cancer genome atlas (TCGA) database. The NRF2- antioxidant response element (ARE) signature was enriched in patients with high TRIB3 expression. Dual-luciferase reporter assay and real-time quantitative polymerase chain reaction (PCR) were used to confirm the effect of TRIB3 on the kelch-like ECH-associated protein-1 (KEAP1)-NRF2 pathway. Abrogation of TRIB3 impaired NRF2 transcriptional activity and reduced the expression of its target genes. Moreover, TRIB3 enhanced NRF2 stability via blocking KEAP1-NRF2 interaction. TRIB3-depletion promoted reactive oxygen species (ROS) production, restrained cell proliferation, and enhanced carboplatin-induced apoptosis. In addition, NRF2 overexpression recovered the tumor inhibition effect of TRIB3-depletion. Consistently, TRIB3 failed to modulate apoptosis in NRF2 depletion cells. In summary, this study shows that TRIB3 inhibits the KEAP1-NRF2 interaction and upregulates the transcriptional activity of NRF2, thereby promoting lung cancer cell proliferation and reducing the sensitivity to chemotherapy. Targeting the TRIB3-NRF2 signal axis may become a new strategy for ROS homeostasis and lung cancer treatment.

Objective To investigate the change of connexin (Cx) in mesenteric artery (MA) and aorta of spontaneously hyper‐tensive rats (SHR) and normotensive rats .Methods Quantitative RT‐PCR and Werstern blot technique were used to compare the difference in the expression of Cx37 and Cx40 mRNA and protein in MA and aorta of SHR and normotensive rats .Results The level of Cx37 mRNA expression in MA from SHR was decreased compared with that of the normotensive rats (P<0 .05) ,and more significantly in aorta (P<0 .01) .Cx40 mRNA expression in MA from SHR was significantly decreased compared with that of the normotensive rats (P<0 .01) ,but no change in aorta (P>0 .05) .The expression of Cx37 protein in MA from SHR was decreased compared with that of the normotensive rats (P<0 .05) ,and more significantly in aorta (P<0 .01) .Cx40 protein expression in MA from SHR was significantly decreased compared with that of the normotensive rats (P<0 .05) ,but no change in aorta (P>0 .05) . Conclusion Hypertension may could decrease gap junctional communication in cells of MA and aorta from SHR by the downregu‐lation of the expression of Cx37 and/or Cx40 .

ObjectiveTo learn the status quo of pre-job training,on-the-job training and the demands of training among community health service staff,and to provide evidence of continuing education.MethodsInvestigation was made by self-designed questionnaire in 335 community health service staff from ten service centers and seven service stations in four regions of Guangdong province.Results The rate of pre-job training and on-the-job training were lower than the training demands among community health service staff.The content of on-the-job training was varied and could meet the training needs.Training was maily done in superior hospitals and the main form of training was seminars and classroom teaching.ConclusionTraining efforts should be increased to meet the training demands.Training model should be innovated to improve the training effect.Hospital and community exchanges should be strengthened and the training system should be improved.

Researching on a large relevant literature of spinal tuberculosis, this paper discussed characteristics of spinal tuberculosis and its early diagnosis method. If spinal tuberculosis can be found and treated earlier, prognosis will be better. The early diagnosis of spinal tuberculosis has become hot spot at present. There are many different diagnostic methods including clinical manifestation, laboratory examination, imaging examination, etc. The newest standpoint showed that immunization has closely correlation with tuberculosis and it will become the focus and developmental direction of future research in spinal tuberculosis. In addition, we should know the differential diagnosis of spinal tuberculosis to avoid the misdiagnosis.
Diagnosis, Differential ; Early Diagnosis ; Humans ; Tuberculosis, Spinal ; diagnosis