Objective To observe the expression change of HRG-1 gene between urinary bladder carcinoma and normal tissues,and to investigate the effect of HRG-1-siRNA on the proliferation and apoptosis of human bladder carcinoma cells.Methods Immunohistochemisty was used to detect the expression of HRG-1 in 85 cases of bladder carcinomas and 20 normal bladder tissues.The siRNA of HRG-1 was designed,synthesized,and transfected into bladder cancer cell line T24.Results The HRG-1 gene expression had significant differences between bladder carcinoma and normal bladder tissues (P ＜ 0.05).The positive expression of HRG-1 gene had significant differences between the pathological grades and clinical stages of bladder carcinomas (P ＜0.05).After treated with siRNA,the expression levels of HRG-1 protein and mRNA in T24 cells decreased obviously (P ＜ 0.05).The apoptosis rate of T24 cells transfected with HRG-1-siRNA was significantly different from control-siRNA group and blank group (P ＜ 0.01).Conclusions The high expression of HRG-1 gene may play an important role in bladder carcinoma,and siRNA targeting HRG-1 can suppress HRG-1 protein expression markedly and enhance apoptosis of T24 cells.

Objective To explore the expression of a new candidate tumor suppressor N-myc downstream regulated gene 2 (NDRG2) in bladder cancer tissues and to investigate its clinical and pathological significance.Methods Formalin-fixed,paraffin-embedded tissue sections from 62 cases of bladder carcinomas and 10 cases of normal bladder tissues were analyzed retrospectively with immunohistochemistry (S-P method).Results The NDRG2 gene was highly expressed in normal bladder tissues,but low expressed in bladder carcinoma tissues.Positive expression of NDRG2 was detected in 8 of the 10 (80.0％) normal tissues and 40.3％ in bladder carcinoma ones (x2 =3.98,P ＜0.05).Furthermore,with the degree of malignancy increased,the positive expression of NDRG2 in bladder carcinoma samples was decreased.The expression of NDRG2 in bladder carcinoma was negatively correlated(r =-0.288,P ＜0.05) with C-myc(r =-0.436,P ＜0.01) and positively correlated with p53 in bladder carcinoma tissues(r =0.717,P ＜0.01).Conclusions The level of NDRG2 expression was lower in bladder carcinomas than in normal tissues.NDRG2 may play an important role in bladder carcinogenesis and in the progress of bladder cancers.

Objective To investigate the antitumor activities of adenovirus-mediated NDRG2 gene (Ad-NDRG2) and docetaxel on human prostate cancer DU145 cells.Methods The protein expressions of cyclin D1,cycliu E,and NDRG2 in the cells were determined by Western blot.MTT and flow cytometry were used to observe the effects of docetaxel (10-6 mol/L,10-7 mol/L,and 10-s mol/L) and Ad-NDRG2 on prostate cancer cell line DU145 in single or synergistic administration ways for 24 and 48 hours in vitro.Male BALB/C-nu mice with DU145 prostate cancer cell lines were treated by docetaxel and Ad-NDRG2 singly or synergistically in vivo.Results After infected by adenovirus,the protein expression of NDRG2 increased,but cyclin D1 and cyclin E decreased in DU145 cells.Ad-NDRG2 inhibited the cell growth (inhibition ratio =41.8％,t =4.18,P ＜0.01),promoted apoptosis (apoptosis ratio =32.4％,x2 =11.66,P ＜0.05),changed the ratio of G2/M phase from 50.2％ to 23.6％,and reversed partially the G2/M arrest,of DU145 cells induced by 10-7 mol/L docetaxel.In vivo experiment showed that docetaxel,Ad-NDRG2,and combination of docetaxel and Ad-NDRG2 inhibited tumor growth with a inhibition rate of 30.7％,28.2％,and 55.8％,respectively.The coefficient of drug interaction (CDI) of docetaxel and Ad-NDRG2 was 0.89.Conclusions Ad-NDRG2 can enhance the growth suppression and apoptosis induced by docetaxel in synergistic way in vitro and in vivo.It demonstrated the great potential of Ad-NDRG2 in the treatment of androgen-independent prostate carcinoma.

Objective To analyze and identify the mutations in SGLT2 gene of nine Chinese families with FRG, and determine the renal threshold for glucose excretion (RTG), so as to explore the association of genotype and RTG. Methods All coding regions of SGLT2 gene, including intron exon boundaries, were analyzed using PCR followed by direct sequence analysis. Quantitative test for 24?hour urine glucose and RTG were measured among 9 probands (21 patients) and their family members from 9 pedigrees (total 25 subjects). The differences in renal glucose thresholds between patients with different genotypes (heterozygotes and compound heterozygotes; c.886(-10_-31) del heterozygotes and other heterozygotes) were compared. Results Twelve mutations were identified by SGLT2 gene analysis, including 10 novel ones that were not included in HGMD:c.331T>C, p.W111R;c.374T>C, p.M125T; c.394C>T, p.R132C; c.612G>C, p.Q204H; c.829C>T, p.P277S; c.880G>A, p.D294N;c.1129G>A, p.G377S; c.1194C>A, p.F398L; c.1540C>T, p.P514S; c.1573C>T, p.H525Y. In thisstudy, the mutation c.886(-10_-31)del that is specific to Chinese population accounted for about 28％of the total alleles (5/18). The RTG values of patients with compound heterozygous mutations were much lower than those with simple heterozygous mutations [(1.28 ±0.10) vs (5.14±0.77) mmol/L; P<0.001];and c.886(-10_-31)del heterozygotes had significant lower RTG values than others [(4.43 ± 0.37) vs (5.70 ± 0.51) mmol/L, P<0.001]. Conclusions Ten novel mutations which may be related to FRG are found in this study, and c.886(-10-31)del may be a hot?spot mutation in Chinese patients. Compound heterozygotes had much lower RTG values than simple heterozygotes.

Objective:To evaluate the availability and safety of laparoscopic pyeloplasty with extracorporeal ureteral tailor.Methods:Clinical data of 26 patients with ureteropelvic junction obstruction (UPJO)who were treated by laparoscopic combined with extracorporeal ureteral tailor pyeloplasty in our hospital from March 2016 to August 2019 were retrospectively analyzed. There were 19 males and 7 females. 22 cases had unilateral lesion, including 6 cases on the right side and 16 cases on the left; 2 cases were bilateral. The average age was 22.3 years old (6-54 years). 19 cases felt discomfort in the renal region and 7 cases were asymptomatic. The mean body mass index was 21.7 kg/m 2 (17.2- 26.4 kg/m 2). 5 cases had mild hydronephrosis, 17 cases had moderate hydronephrosis and 4 cases had severe hydronephrosis. 6 cases combined with cross vessels. In all the 26 cases, the ureter was pulled out of the abdomen through a laparoscopic incision, cut lengthways in vitro, sutured at the lowest point, and then returned it to the abdomen. Then, double J tube implantation and ureteropelvic anastomosis were performed under the laparoscopy. Results:26 cases were completed successfully without conversion, with the average operation time of 99 minutes (50-158 minutes), the average blood loss of 19.4 ml (10-50 ml), the average hospital stay of 6.5 days (5-11 days), and the average drainage indwelling time of 5.3 days (4-10 days). For complications, urine leakage occurred on the 3rd day after the operation in 1 case, and the daily drainage fluid was more than 500 ml, which decreased suddenly after 4 days. Postoperative average follow-up was 10.8 months (6-24 months). Renal region pain disappeared in all patients. CT reexamination 3 months after the operation showed that hydronephrosis was alleviated or disappeared in 24 cases, and there was no significant change in 2 cases compared with the preoperative images, but no progress was found in the subsequent reexamination, so we didn't deal with the hydronephrosis again.Conclusions:Laparoscopic pyeloplasty combined with extracorporeal ureteral tailor is minimally invasive and flexible, which greatly reduces the difficulty and time of operation, and has a high success rate.

Objective:The purpose of this study was to investigate the expression and role of N-myc downstream regulatory gene 2 (NDRG2) in radiation resistance of bladder cancer cells.Methods:T24 cells were cultured in vitro and irradiated with different doses of X-ray (0, 2, 4, 8, 10 and 20 Gy). The best dose of X-ray was selected for subsequent treatment. The radioresistant BCa cell line T24/R was established. The cytotoxicity of T24/R cells was detected by counting kit-8 (CCK-8) method. The proliferation and invasion ability of T24/R cells and T24 cells were detected by flow cytometry and transwell, respectively. Western blot was used to detect the expression of epithelial mesenchymal transition (EMT) related proteins. The survival rate of T24/R group (control group) and T24/R-NDRG 2 group was detected, and the migration ability of T24/R-NDRG 2 cells was detected after 2 Gy treatment. Results:The cell viability was inhibited significantly when the dose of X-ray was ≥2 Gy X-ray, so 2 Gy X-ray irradiation was chosen as the best condition for BCa cytotoxicity and T24/R radiation resistance cell line was successfully established; Apoptosis test showed that the number of S-phase cells was increased in T24/R group, and the proportion of S-phase cells in T24/R vs T24 was (26.49±4.5)% vs (14±2.6)% ( P<0.05); Transwell test showed that T24/R cells showed stronger migration ability than control group ( P<0.05), but there was no significant difference in EMT related protein expression between the two groups ( P>0.05). Overexpression of NDRG2 can significantly decreased the activity and migration ability of radiation-resistant T24/R cells ( P<0.05) when the radiation dose was gradually increasing in both groups. Conclusions:The radiation resistance of BCa cells is one of the causes of local tumor recurrence. Up-regulation of NDRG2 expression can inhibit the radiation resistance of T24 cells, so it can be used as a candidate for treatment of radiation-resistant BCa patients.

Objective To compare the etiological constitution of recurrent miscarriage (RM) between patients with consecutive two and three or more miscarriages through combining the routine examination results and embryonic karyotype. Methods Patients with a history of two or more consecutive clinical miscarriages(≤12 weeks of gestation)consulting in the RM clinic of the First Affiliated Hospital of Sun Yat-sen University from March 2011 to January 2016 were collected. Six hundred and ninety-six with detailed history recorded, routine clinical examinations of RM and at least once embryonic karyotype were ultimately enrolled in this study. Their etiological constitution of RM were analyzed in groups of consecutive two and three or more miscarriage. The etiologies of RM in analysis consisted of women age, body mass index (BMI), chromosome abnormalities of couples, uterine abnormalities, endocrinology abnormalities and antiphospholipid syndrome(APS). Results (1)Among 696 patients, the abnormal embryonic karyotypes was 60.6%(422/696)and routine RM etiologies was 32.2%(224/696), leaving the ratio of unexplained RM was only 29.0%(202/696).(2)A total of 717 embryo karyotype were found in 696 patients, included 21 cases with twice embryo karyotype results the percentage of normal embryo was 39.7%(285/717), while abnormal ones was 60.3%(432/717). Among the types of abnormal karyotype, the most common ones (>10%)were trisomy 16(19.2%, 83/432), monosome X(11.3%, 49/432)and trisomy 22(10.9%, 47/432). (3)Among the 696 RM patients, the number of two and three or more miscarriages were respectively 446(64.1%,446/696)and 250(35.9%,250/696). Comparing groups of three or more miscarriages with two miscarriages, there were significant differencein older age as well as uterine adhesion(P<0.05). But no difference was found in body mass index(BMI), the rates of chromosome abnormalities of couples, uterine abnormalities except uterine adhesion, endocrinology abnormalities and APS (all P>0.05) between two groups. Conclusions The abnormal embryonic karyotype is the most common cause of first-trimester RM. The etiological constitution of two and three or more recurrent miscarriages is accordant, suggesting that routine clinical examination and the embryonic karyotype should be started following two consecutive clinical early miscarriages.

Objective:To investigate the predictive value of preoperative blood parameters on the efficacy of microscopic varicocele ligation in the treatment of infertility patients with varicocele.Methods:The clinical data of 110 patients with varicocele admitted to Xi′an People′s Hospital and Tangdu Hospital of Air Force Military Medical University from June 2016 to January 2021 were analyzed retrospectively. Successful operation was defined as the restoration of all semen parameters (sperm concentration, forward motility, morphology, etc.) to normal values 6 months after operation. The patients were divided into effective treatment group (group A, 78 cases) and ineffective treatment group (group B, 32 cases). The blood parameters of the two groups were compared before operation. Spearman correlation analysis was used to determine the relationship between blood parameters [neutrophil/lymphocyte ratio (NLR) and mean platelet volume (MPV)] and the success of microscopic varicocele ligation. The predictive value of blood parameters (NLR and MPV) to the success of microscopic varicocele ligation was analyzed by receiver operating characteristic (ROC) curve.Results:There was no significant difference in clinical characteristics, and sex hormone levels between the two groups before operation (all P>0.05); There were statistically significant differences in NLR and MPV in blood parameters between the two groups (both P<0.05), and there was no significant differences in other blood parameters (both P>0.05). There was no significant difference in semen parameters between the two groups before operation (all P>0.05). After operation, the semen volume, semen concentration, forward movement and sperm morphology of patients in group A were significantly improved compared with those before operation (all P<0.05), while the parameters of patients in group B had no significant difference compared with those before operation (all P>0.05). There was a negative correlation between preoperative NLR and the success rate of spermatic vein ligation ( r=-0.719, P<0.01), and a positive correlation between MPV and the success rate of spermatic vein ligation ( r=0.522, P<0.01). The ROC curve was used to analyze the threshold of predictive variables for the success of spermatic vein ligation. The optimal critical value of NLR was 2.01 ( P<0.01), and the optimal critical value of MPV was 11.45 ( P<0.01). Conclusions:Low NLR (<2.01) and high MPV (>11.45) may be useful preoperative predictive tools for identifying the group of infertile varicocele patients who would benefit most from microscopic spermatic vein ligation.

Objective:To investigate the role of miR-144-3p in cisplatin resistance of bladder cancer.Methods:Bladder cancer T24 cells were cultured in vitro and divided into blank group (untreated), mimetic control group, miR-144-3p mimetic transfection group, inhibitor control group, and miR-144-3p inhibitor transfection group. Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to verify the transfection effect, methyl thiazolyl tetrazolium (MTT) method was used to detect the survival rate of cells treated with cisplatin in each group, and Western blot was used to detect the expression of the target protein. The targeting relationship between miR-144-3p and nuclear factor E2 related factor 2 (Nrf2) was validated using dual fluorescence reporter gene experiments. Furthermore, Nrf2 was knocked out in each group of cells, and the mRNA and protein expression levels of HO-1, Bcl-2, and Caspase-3 were detected by qRT-PCR and Western blot in each group of cells.Results:Compared with the control group, bladder cancer cells in the miR-144-3p mimetic transfection group were more sensitive to cisplatin, while the miR-144-3p inhibitor transfection group had the opposite effect; The miR-144-3p simulant transfection group can effectively inhibit the mRNA and protein expression level of Nrf2 in bladder cancer cells (all P<0.05), while the miR-144-3p inhibitor transfection group can up regulate the mRNA and protein level of Nrf2 (all P<0.05). The miR-144-3p mimetic transfection group showed significant downregulation of mRNA and protein expression of HO-1 and Bcl-2, while the expression of Caspase-3 was upregulated (all P<0.05), while the miR-144-3p inhibitor transfection group showed the opposite results. The luciferase results confirmed that miR 144 3p can directly bind to the 3′- UTR region of Nrf2, reducing the mRNA level of Nrf2. When Nrf2 was knocked out, whether miR-144-3p mimetic transfection group or miR-144-3p inhibitor transfection group, the mRNA and protein expression levels of HO-1, Bcl-2 and Caspase-3 did not change significantly, and miR-144-3p lost the ability to regulate the cisplatin sensitivity of bladder cancer cells. Conclusions:miR-144-3p targets to regulate the sensitivity of Nrf2 to cisplatin in bladder cancer, and miR-144-3p is expected to become a new target for the treatment of cisplatin resistant or refractory bladder cancer.

Objective: To investigate the Rhizoma Pleionis mediating vascular endothelial growth factor A (VEGF-A)-related angiogenesis in breast cancer cells to inhibit epithelial-mesenchymal transition (EMT) in breast cancer. Methods: Firstly, the proliferation activity of MDA-MB-231 and 4T1 of breast cancer cells was detected by methyl thiazolyl tetrazolium (MTT) method at different concentrations (0.2 g/ml, 0.4 g/ml, 0.6 g/ml, 0.8 g/ml, 1.0 g/ml); secondly, the specific concentration of Rhizoma Pleionis was selected and tested by Transwell test for its effect on breast cancer cell metastasis and invasion. The formation of human umbilical vein endothelial cells (HUVEC) capillary wall was measured to detect the effect of Rhizoma Pleionis on cell angiogenesis. The EMT-related protein and angiogenesis-related molecules were detected by Western blot. Results: MTT results showed that Rhizoma Pleionis of 0.8 g/ml and 1.0 g/ml can inhibit the MDA-MB-231 and 4T1 cells proliferation in a dose-dependent manner, with statistically significant difference (P<0.05). Rhizoma Pleionis can inhibit the invasion of breast cancer cells and inhibit the formation of blood vessel walls in HUVECs. Western blot confirmed that the expression level of EMT-related proteins: E-cadherin was significantly increased and the expression level of N-cadherin, Vimentin and Snail was significantly decreased after treatment with Rhizoma Pleionis (P<0.05); and the level of the related molecule hypoxia inducible factor-1α (HIF-1α), VEGF-A and vascular endothelial growth factor receptor-2 (VEGFR-2) was significantly reduced (P<0.05). Conclusions Rhizoma Pleionis inhibits the epithelial-mesenchymal transition of breast cancer by influencing the angiogenesis of VEGF-A, and inhibits the invasion and metastasis of breast cancer.