Objective To evaluate microdeletion of DAZ gene on the Y chromosome in patients suffering from idiopathic azoospermia or severe oligozoospermia.Methods The four sites including SY154,SY155,SY254,SY255 and SRY regions of Y chromosome in 38 cases of idiopathic azoospermia and severe oligozoospermia were detected by STS-PCR.Results Six cases of microdeletion of DAZ genes were found in 38 patiens,microdeletion rate was 15.8%.Five cases from 30 patiens suffering from idiopathic azoospermia,microdeletion rate was 16.7%.One case from 8 patients suffering from severe oligozoospermia had microdeletion.The microdeletion rate was 12.5%.Conclusions The microdeletion of DAZ gene is a major cause of idiopathic azoospermia and severe oligozoospermia leading to male infertility.Therefore,DAZ gene should be detected and PGD should be done prior to ICSI for the patients suffering from idiopathic azoospermia and severe oligozoospermia.

Objective The purpose of this paper is to study the relationship between blood concentration and brain tissue drug concentration by different dose of TMX chemotherapy acute lymphoblastic leukemia in mice. Methods 4 weeks, health Kun Ming mice 80: establishment acute lymphoblastic leukemia mice model,20 mice were randomly selected to take the femur bone marrow biopsy bone marrow OK for model verification; the remaining 60 acute lymphoblastic leukemia mice were allocated randomly 6 groups of 10 mice in each group, respectively A, B, C, D, E, F groups. And collected blood 0.5 ml and brain tissue 0.4 g individually at 0.5 hour in every group. We used supernatant of centrifugation blood and brain homogenate to detected drug concentration by fluorescence polarization immunoassay. Results The mean blood concentration of MTX of six groups A, B, C, D, E, F are (39.08±5.18) μmol/L, (15.86±1.02)μmol/L, (8.67± 5.43)μmol/L, (68.29±5.19)μmol/L, (29.55±6.22)μmol/L, (13.98±1.12)μmol/L, respectively. Compared the mean blood concentration of MTX of each group there are statistical significance (P<0.05). The mean concentration of MTX of six groups in brain tissue are followed by A group (1.05±0.26)μmol/L, B group (0.61±0.25)μmol/L, C group (0.48±0.25)μmol/L, D group (2.07±0.35)μmol/L, E group (1.27±0.21)μmol/L, F group (0.59±0.69)μmol/L. Compared the mean concentration of MTX of each group in brain tissue there are statistical significance (P<0.05). MTX concentration in blood and in brain tissue of correlation coefficient followed by 0.82, 0.75, 0.19, 0.81, 0.55, 0.43. Conclusion The chemotherapy acute lymphoblastic leukemia mice of HDMTX scheme, the peak of blood concentration and brain tissue drug concentration is come after injected MTX 0.5 hour, MTX 5 g/m~2 is better permeation blood-brain barrier and more easy make brain tissue drug concentration to reach effectively therapeutic concentration than MTX 3 g/m~2.

Objective To explore the application value of minimally invasive surgery in the treatment of lumbar spinal tuberculosis, and to provide reference for clinical treatment of spinal tuberculosis. Methods Data of 252 cases of patients with lumbar spinal tuberculosis treated by conservative treatment in our hospital from January 2005 to December 2014 were retrospectively analyzed. Patients were divided into four groups on the basis of systemic application of antituberculosis chemotherapy. A total of 154 patients were given simple local chemotherapy of percutaneous placement of focus catheter (group A), 48 patients were received percutaneous perfusion drainage and local chemotherapy (group B), 32 patients underwent percutaneous puncture catheter debridement combined with local chemotherapy (group C), and 18 patients were given percutaneous debridement and internal fixation combined with local chemotherapy catheter (group D). Data of erythrocyte sedimentation rate (ESR), visual analogue scale (VAS), Oswestry disability index (ODI) score and the modified MacNab criteria were recorded before operation and at the end of the follow-up in four groups of patients. Results Of the 252 patients, 228 were followed up and 214 patients achieved clinical cure. The lost access were15 cases in group A, 5 cases in group B, 2 cases in group C and 2 cases in group D. The total rate of lost visit was 9.52%. The follow up duration ranged from 25-126 months. The mean duration of follow-up was 68(60, 76) months. A total 214 cases reached the standard of clinical cure. No complications (retrograde infection and cross infection) were found in all patients during treatment. ESR was statistically decreased to (7.26 ± 3.43) mm/1 h at the last follow-up (t=35.06, P=0.023) compared with that (44.96 ± 12.42) mm/1 h before operation. The VAS and ODI were 1.5(1, 3) and 30(25, 35)% at the last follow-up, which were significantly improved than those [7.5(7.0, 8.0) and 60(55, 65)%] before operation (Z=13.641 and 6.806, P<0.05). According to the improved MacNab criteria, the overall excellent and good rates for patients were 86.4%(197/228) at the last follow-up. Conclusion According to the stepped care and personalized treatment, patients of lumbar tuberculosis are preoperative comprehensive evaluated, and most patients can achieve long-term stability and a better clinical efficacy after interventional and minimally invasive treatment.

Objective:To investigate the clinical efficacy of partial excision of nail matrix combined with phenolic ablation in the treatment of ingrowing toenail.Methods:115 patients(148 toenails) with ingrowing toenail treated in the Central hospital of Wuhan from October 2004 to May 2013 were selected for this study.The patients were divided into two groups,53 patients(67 toenails) admitted from October 2004 to December 2007 were considered as the observation group and treated with Partial excision of nail matrix,62 patients(81 toenails) admitted from January 2008 to May 2013 were considered as the control group and treated with Partial excision of nail matrix and phenolic ablation.The bleeding time,pain relief time,healing time,recurrences after one year and satisfaction rate of two groups were compared.Results:The wounds of 148 toenails were healing.The Bleeding time,pain relief time,recurrences after one year in the control group (1.85:±:0.42days,13.25± 2.17hours) were lower than the observation group (2.69± 0.53 days,21.54± 2.56hours),and healing time in the control group (11.32± 2.37days) were longer than the observation group (8.93± 2.06 days)(P＜0.05),the recurrence rate and overall satisfaction rate of observation group and control group were 6.15％,12.5％ and 97.06％,91.07％.Conclusion:Partial excision of nail matrix combined with phenolic ablation was more effective in the treatment of ingrowing toenail than surgical excisional techniques.

Objective:To detect the expression levels of neuropilin1 (NRP1)mRNA and miR-9 in non-small cell lung cancer (NSCLC)tissue samples, and to explore the correlations between the expressions of NRP1 mRNA, miR-9 and the clinicopathological characteristics of the patients with NSCLC.Methods:Informed consent was obtained from each patient before surgery.The tissue samples including 45 NSCLC tissue ,45 adjacent carcinoma tissue and 45 normal lung tissue were collected from China-Japan Union Hospital of Jilin University from 2010 to 2011.qRT-PCR was used to detect the expression levels of NRP1 mRNA and miR-9 in three kinds of lung tissue, and the correlation between the expressions of NRP1 mRNA, miR-9 and clinicopathological characteristics of the patients with NSCLC was analyzed.Results:Compared with normal tissue,the expression level of NRP1 mRNA in adjacent carcinoma tissue had no change (P>0.05),but the expression level of NRP1 mRNA in non-small cell lung cancer tissue was significantly decreased (P<0.05).Compared with normal tissue,the expression level of miR-9 in adjacent carcinoma tissue had no change (P>0.05),but the expression level of miR-9 in non-small cell lung cancer tissue was significantly increased (P < 0.05 ). Furthermore, in adjacent carcinoma tissue, the expression level of miR-9 in the males was lower than that in the females (P<0.05 ). In NSCLC tissue, the expression level of NRP1 mRNA had no relationship with sex,age,differentiation degree,TNM stage and clinical stage,but was significantly correlated to the histological subtype and lymph node metastasis (P<0.05).In NSCLC tissue,the expression level of miR-9 had no relationship with age, pathological type, lymph node metastasis, differentiation degree,TNM stage,and clinical stage (P>0.05),but was correlated to the sex (P<0.05). Conclusion:The expression level of miR-9 is up-regulated and the expression level of NRP1 mRNA is down-regulated significantly in non-small cell lung cancer tissue. The detection of the expression level of NRP1 mRNA contributes to j udge the histological subtype and lymph node metastasis of NSCLC.

Objective To analyze the clinical effects of arthroscopic en masse repair with footprint ending shift using double-row suture-bridge technique for delaminated rotator cuff tears under tension.Methods A total of 58 patients with delaminated rotator cuff tears under tension from August 2013 to August 2016 who underwent arthroscopic en masse repair using doublerow suture-bridge technique were retrospectively analyzed.There were 33 males and 25 females with a mean age of 53.0±7.8 years (range 39-74) with 24 patients left side involved and 34 right side.They were divided into 2 groups to receive en masse repair either footprint ending shift or on the footprint.There were 28 patients with footprint ending shift and 30 patients on the footprint.Clinical effects were evaluated by University of California Los Angeles (UCLA) score,American Shoulder and Elbow Surgeons (ASES) score,visual analogue scale (VAS),Constant-Murley score and shoulder range of motion at preoperatively and postoperatively.Results The average follow-up duration was 23.2±0.8 months (range 21-24).The two groups were compatible with no significant difference in age,gender,tear size,follow-up duration,preoperative function and range of motion of the shoulder joint (P＞0.05).At the last follow up,the UCLA,ASES,VAS,Constant-Murley scores and shoulder range of motion in the group footprint ending shift were respectively 32.4±2.5,12.8±0.9,1.0±1.1,93.4±5.6,158.3°±9.3°,58.9°±5.0° with significantly differences compared with preoperative scores (P ＜ 0.05).The postoperative value in the group on footprint were respectively 31.6±2.9,12.8±0.9,0.7 ± 1.2,91.3±7.1,156.1°± 10.7°,59.6°±4.6° with significantly differences compared with the preoperative scores (P ＜ 0.05).There were no significant difference between the two groups (P ＞ 0.05).The operation duration in the group footprint ending shift was 100.9±6.0 min,while that in the group on footprint was 106.6±6.1 min.There was significantly difference in the operation duration between two groups (t=-3.600,P=0.001).Conclusion Arthroscopic en masse repair using double-row suture-bridge technique can successfully treat delaminated rotator cuff tears under tension.Compared with arthroscopic en masse repair on footprint using double-row suture-bridge technique,the footprint ending shift is easier and time saving without significant difference in function of the shoulder joint and the range of motion in repair of delaminated rotator cuff tear under tension.

Objective To evaluate the clinical efficacy of arthroscopic bursal layer-only double-row suture-bridge repair for delaminated rotator cuff tear which is difficult to reposit in comparison with separate double-layer repair and whole-layer repair.Methods From May 2013 through June 2016,82 patients with delaminated rotator cuff tear difficult to reposit were treated at Department of Joint Surgery,The Affiliate Hospital to Chengde Medical University.They were 47 males and 35 females with a mean age of 53.0 ± 7.9 years.They were divided into 3 groups according to their surgical procedures.In group A,28 cases were treated by arthroscopic whole-layer double-row suture-bridge procedure;in group B,29 cases were treated by arthroscopic separate double-layer double-row suture-bridge procedure;in group C,25 cases were treated by arthroscopic bursal layer-only double-row suture-bridge procedure.The 3 groups were compared in terms of University of California Los Angeles (UCLA) score,American Shoulder and Elbow Surgeons (ASES) score,visual analogue scale (VAS),Constant shoulder score,range of motion of shoulder joint and rotator cuff retear preoperatively and postoperatively.Results The patients in the 3 groups were comparable because their preoperative general data showed no significant significances (P ＞ 0.05).The operation time for groups A,B and C was respectively 105.5 ±5.6 min,117.4 ±6.9 min and 88.0 ±4.2 min,showing significant differences between the 3 groups (P ＜ 0.05).The 82 patients were followed up for 21 to 24 months (average,23.3 months).At 24 months postoperatively,UCLA,ASES,VAS,Constant score,shoulder anteflexion and lateral extorsion were respectively 32.4 ± 2.5,12.8 ± 0.9,1.0 ± 1.1,93.4 ± 5.6,158.3° ± 9.3°and 58.9°±5.0°in group A,32.2±2.5,12.9±1.0,0.9±1.0,92.8±6.0,156.4°±9.5°and 59.3°± 5.6° in groups B,and32.4±2.4,12.9±0.9,0.7±0.9,94.3±5.2,156.0°±9.5°and57.6°°5.4°in group C,showing no significant differences between the 3 groups (P ＞ 0.05).The occurrence of rotator cuff retear in groups A,B and C were respectively 17.9％ (5/28),13.8％ (4/29) and 12.0％ (3/25),showing no significant differences between the 3 groups (P ＞ 0.05).Conclusions In repair of delaminated rotator cuff tear difficult to reposit,although the arthroscopic bursal layer-only double-row suture-bridge repair is similar to conventional arthroscopic whole-layer double-row suture-bridge repair and arthroscopic separate double-layer double-row suture-bridge repair in functional recovery and range of motion of the shoulder and incidence of rotator cuff retear,it can reduce obviously operation time and make the operation easier.

Previous studies have found that As2 O3 can interfere with serum thyroid hormone TH levels in rats and cause chronic liver damage,but the mechanism remains unclear.In order to ex-plore the role of TH signaling pathway in As2 O3-induced chronic liver injury,qRT-PCR and West-ern blot techniques were used to detect the expression changes of genes and protein of TRβ1(a key regulator of TH signaling pathway in rat liver)and cyclin D1(the downstream factor of TRβ1 in nuclear pathway).Meanwhile,the changes in the protein of key factors(Bax,Bcl-2)of the TH sig-nal nuclear outside pathway were detected.The results indicated that:after As2 O3 treatment for 110 days,compared with the control group,the expression of TRβ1 protein in the liver of female mice significantly decreased(P＜0.01),the expression of cyclin D1 significantly increased in the 0.1 and 0.2 mg/L groups(P＜0.01).Meanwhile,the expression of TRβ1 protein in male mice sig-nificantly decreased in 0.4 mg/L group(P＜0.01),and the expression of cyclin D1 in each group significantly increased(P＜0.01).The mRNA expression results were basically the same as those of protein expression.After As2 O3 treatment for 194 days,compared with the control group,the expression of TRβ1 protein in each group significantly decreased(P＜0.01),and the expression of cyclin D1 significantly increased(P＜0.01).The mRNA expression results were basically consist-ent with the protein.As2 O3 interfered with the expression of Bcl-2 and Bax proteins in rats and in-duced the increase in the ratio of Bcl-2/Bax protein as the action time increased.Among them,the Bcl-2/Bax ratio of female rats in each group and male rats in the 0.4 mg/L group significantly in-creased(P＜0.01),and male rats in the 0.1 mg/L group significantly increased(P＜0.05).It shows that As2O3 can cause abnormal levels of TRβ1,cyclin D1 and the Bcl-2/Bax ratio in rat liv-er.

Objective:To investigate the effect and mechanism of SIRT7 in epithelial mesenchymal transformation (EMT) of pancreatic cancer cells.Methods:The pancreatic cancer cells were divided into siControl, siSIRT7, over-expression SIRT7, siSIRT7+siCOL4A1, and siSIRT7+siSLUG groups using siRNA or plasmid transfection. The proliferation, migration and invasion of pancreatic cancer cells were detected by EdU, wound healing assay and Transwell experiments, respectively. The expression of EMT and cancer stem cell (CSC) markers were detected by quantitative real-time reverse transcription polymerase chain reaction assay (qRT-PCR) and western blot. RNA sequencing (RNA-seq) in SIRT7 knockdown PANC-1 cells was performed to explore the signaling pathways and target genes regulated by SIRT7. Then the target genes directly regulated by SIRT7 were identified with quantitative chromatin immunoprecipitation experiment (q-ChIP) and chromatin immunoprecipitation polymerase chain reaction (ChIP-PCR). The expressions of SIRT7 and target genes were detected by immunohistochemical (IHC) in pancreatic cancer tissues, and the correlation between SIRT7 and target gene expression was analyzed using TCGA dataset. The correlation between expression of SIRT7 or target genes and survival was analyzed on KM-plotter website. Finally, GeneMANIA, STRING and ENCORI were used to predict SIRT7-related proteins and miRNAs.Results:EdU assay showed that the cell proliferation rates in SIRT7-overexpressed PANC-1 [(19.33±0.35)%] and BxPC-3 cells [(17.00±1.89)%] were lower than those in the control group [(31.60±1.37)% and (24.33±0.78)%, respectively, P＜0.05]. The proliferation rates of SIRT7-knockdown PANC-1 [(23.94±1.00)% and (27.08±0.97)%] and BxPC-3 cells [(22.00±1.86)% and (25.96±1.61)%] were higher than those of the siControl group [(11.80±1.86)% and (13.42±1.39)%, respectively, P＜0.05]. In PANC-1 cells, the wound healing assay showed that the relative migration rate of SIRT7-overexpression cells [(76.67±2.74)%] was lower than that of control cells [(100.00±2.13)%, P＜0.05]; the relative migration rate of cells with SIRT7 knockdown [(134.22±4.08)% and (199.82±9.20)%, respectively] was higher than that of siControl group [(102.24±3.13)%, P＜0.05]. Compared with the control group, SIRT7 overexpression decreased the number of migrated BxPC-3 cells (45.66±1.69 vs 28.33±2.62, P＜0.05); while SIRT7 knockdown increased these numbers (65.66±2.86 and 82.00±2.94 versus 33.00±0.81, P＜0.01). Transwell experiment revealed that the number of invaded cells in SIRT7 overexpression groups (16.33±2.05 and 34.66±1.69) was lower than that control groups (54.33±4.64 and 58.66±5.90, P＜0.05); with SIRT7 knockdown, the numbers of invaded PANC-1 (63.66±2.49 and 69.33±3.29) and BxPC-3 cells (134.33±3.09 and 181.66±4.02) were higher than those in control groups (35.33±2.49 and 42.00±0.81, P?0.05). Also, SIRT7 knockdown decreased the expressions of epithelial markers and increased the expressions of mesenchymal and CSC markers. RNA-seq analysis showed that SIRT7 was involved in regulating a variety of cancer-related signaling pathways, including the pancreatic cancer pathway and the EMT pathway. Furthermore, SIRT7 could directly bind to the promoter regions of target genes, such as COL4A1 and SLUG. SIRT7 was negatively correlated with the expression and function of COL4A1 and SLUG in pancreatic cancer cells. The expressions of SIRT7, COL4A1, SLUG and SOX2 were verified in pancreatic cancer tissues by IHC. Finally, SIRT7 was predicted to be associated with many proteins and miRNAs based on GeneMANIA, STRING, and ENCORI online tools. Conclusions:SIRT7 can inhibit the EMT of pancreatic cancer cells through transcriptionally inhibiting the expression of target genes, such as COL4A1 and SLUG. Thus, SIRT7 may serve as a potential tumor suppressor gene in pancreatic cancer.

Objective:To investigate the effect and mechanism of SIRT7 in epithelial mesenchymal transformation (EMT) of pancreatic cancer cells.Methods:The pancreatic cancer cells were divided into siControl, siSIRT7, over-expression SIRT7, siSIRT7+siCOL4A1, and siSIRT7+siSLUG groups using siRNA or plasmid transfection. The proliferation, migration and invasion of pancreatic cancer cells were detected by EdU, wound healing assay and Transwell experiments, respectively. The expression of EMT and cancer stem cell (CSC) markers were detected by quantitative real-time reverse transcription polymerase chain reaction assay (qRT-PCR) and western blot. RNA sequencing (RNA-seq) in SIRT7 knockdown PANC-1 cells was performed to explore the signaling pathways and target genes regulated by SIRT7. Then the target genes directly regulated by SIRT7 were identified with quantitative chromatin immunoprecipitation experiment (q-ChIP) and chromatin immunoprecipitation polymerase chain reaction (ChIP-PCR). The expressions of SIRT7 and target genes were detected by immunohistochemical (IHC) in pancreatic cancer tissues, and the correlation between SIRT7 and target gene expression was analyzed using TCGA dataset. The correlation between expression of SIRT7 or target genes and survival was analyzed on KM-plotter website. Finally, GeneMANIA, STRING and ENCORI were used to predict SIRT7-related proteins and miRNAs.Results:EdU assay showed that the cell proliferation rates in SIRT7-overexpressed PANC-1 [(19.33±0.35)%] and BxPC-3 cells [(17.00±1.89)%] were lower than those in the control group [(31.60±1.37)% and (24.33±0.78)%, respectively, P＜0.05]. The proliferation rates of SIRT7-knockdown PANC-1 [(23.94±1.00)% and (27.08±0.97)%] and BxPC-3 cells [(22.00±1.86)% and (25.96±1.61)%] were higher than those of the siControl group [(11.80±1.86)% and (13.42±1.39)%, respectively, P＜0.05]. In PANC-1 cells, the wound healing assay showed that the relative migration rate of SIRT7-overexpression cells [(76.67±2.74)%] was lower than that of control cells [(100.00±2.13)%, P＜0.05]; the relative migration rate of cells with SIRT7 knockdown [(134.22±4.08)% and (199.82±9.20)%, respectively] was higher than that of siControl group [(102.24±3.13)%, P＜0.05]. Compared with the control group, SIRT7 overexpression decreased the number of migrated BxPC-3 cells (45.66±1.69 vs 28.33±2.62, P＜0.05); while SIRT7 knockdown increased these numbers (65.66±2.86 and 82.00±2.94 versus 33.00±0.81, P＜0.01). Transwell experiment revealed that the number of invaded cells in SIRT7 overexpression groups (16.33±2.05 and 34.66±1.69) was lower than that control groups (54.33±4.64 and 58.66±5.90, P＜0.05); with SIRT7 knockdown, the numbers of invaded PANC-1 (63.66±2.49 and 69.33±3.29) and BxPC-3 cells (134.33±3.09 and 181.66±4.02) were higher than those in control groups (35.33±2.49 and 42.00±0.81, P?0.05). Also, SIRT7 knockdown decreased the expressions of epithelial markers and increased the expressions of mesenchymal and CSC markers. RNA-seq analysis showed that SIRT7 was involved in regulating a variety of cancer-related signaling pathways, including the pancreatic cancer pathway and the EMT pathway. Furthermore, SIRT7 could directly bind to the promoter regions of target genes, such as COL4A1 and SLUG. SIRT7 was negatively correlated with the expression and function of COL4A1 and SLUG in pancreatic cancer cells. The expressions of SIRT7, COL4A1, SLUG and SOX2 were verified in pancreatic cancer tissues by IHC. Finally, SIRT7 was predicted to be associated with many proteins and miRNAs based on GeneMANIA, STRING, and ENCORI online tools. Conclusions:SIRT7 can inhibit the EMT of pancreatic cancer cells through transcriptionally inhibiting the expression of target genes, such as COL4A1 and SLUG. Thus, SIRT7 may serve as a potential tumor suppressor gene in pancreatic cancer.