Objective To investigate the role of RNA-binding motif protein X-linked(RBMX)in regulating the proliferation,migration,invasion and glycolysis in human bladder cancer cells.Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown,respectively,and successful cell modeling was verified using RT-qPCR and Western blotting.Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony-forming assay,and cell migration and invasion abilities were determined using Transwell experiment.The expressions of glycolysis-related proteins M1 pyruvate kinase(PKM1)and M2 pyruvate kinase(PKM2)were detected using Western blotting.The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits.Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown.RBMX overexpression significantly inhibited the proliferation,clone formation,migration and invasion of bladder cancer cells,while RBMX knockdown produced the opposite effects.Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2,while RBMX knockdown produced the opposite effects.Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression(P<0.05)but increased significantly following RBMX knockdown(P<0.05).Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression,suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

Objective To investigate the role of RNA-binding motif protein X-linked(RBMX)in regulating the proliferation,migration,invasion and glycolysis in human bladder cancer cells.Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown,respectively,and successful cell modeling was verified using RT-qPCR and Western blotting.Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony-forming assay,and cell migration and invasion abilities were determined using Transwell experiment.The expressions of glycolysis-related proteins M1 pyruvate kinase(PKM1)and M2 pyruvate kinase(PKM2)were detected using Western blotting.The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits.Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown.RBMX overexpression significantly inhibited the proliferation,clone formation,migration and invasion of bladder cancer cells,while RBMX knockdown produced the opposite effects.Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2,while RBMX knockdown produced the opposite effects.Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression(P<0.05)but increased significantly following RBMX knockdown(P<0.05).Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression,suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

Objective:To explore the effect of the developmental stages and quality on pregnancy outcome and birth outcome, and provide evidence for single blastocyst selection in frozen-thawed cycles.Methods:A retrospective cohort study analysis was performed on the data of patients with a total of 893 cycles who underwent single blastocyst transfer in frozen-thawed cycles in the Center for Reproductive Medicine, Qingyuan People's Hospital from January 2013 to June 2021. The cycles were divided into day 5 (D5) and day 6 (D6) groups according to the time of blastocyst formation. Then the two groups were divided into four subgroups according to the quality of blastocyst, namely, D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup. The general data, clinical outcomes and neonatal outcomes of each group were compared.Results:1) The clinical pregnancy rate [60.14% (332/552)], the implantation rate [60.14% (332/552)] and the live birth rate [47.64% (263/552)] in D5 group were significantly higher than those in D6 group [45.75% (156/341), 45.75% (156/341), 36.36% (124/341), all P<0.001], but there were no significant differences in body mass index, duration of infertility, intimal thickness of transplantation day and miscarriage rate between the two groups (all P>0.05). In addition, there were also no significant differences in birth weight, low birth weight rate, fetal macrosomia rate and male/female ratio (all P>0.05). 2) There were significant differences in clinical pregnancy rate [61.00% (294/482), 54.29% (38/70), 51.00% (127/249), 31.52% (29/92)] and live birth rate [48.96% (236/482), 38.57% (27/70), 41.37% (103/249), 22.83% (21/92)] among D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup (all P<0.001). D5 good-quality embryo subgroup had the highest clinical pregnancy rate and live birth rate, while D6 non-good-quality embryo subgroup had the lowest clinical pregnancy rate and live birth rate. There were also no significant differences in birth weight, fetal macrosomia rate and male/female ratio among the four subgroups (all P>0.05), while there was a significant difference in low birth weight rate [5.08% (12/236), 0 (0/27), 4.85% (5/103), 23.81% (5/21)] among the four subgroups ( P=0.014). 3) There were no significant differences in clinical pregnancy rate and live birth rate between D5 non-good-quality embryo subgroup and D6 good-quality embryo subgroup (all P>0.05). The clinical pregnancy rate and the live birth rate of 4BC in D5 were lower than those of 4AA, 4AB and 4BA in D6, while the miscarriage rate of 4BC in D5 was higher than that of 4AA, 4AB and 4BA in D6, but there were no significant differences (all P>0.05).The clinical pregnancy rate and the live birth rate of 4BC in D5 were higher than those of 4BB in D6, but there were no significant differences (all P>0.05). Conclusion:In the frozen-thawed cycle of single blastocyst transplantation, D5 good-quality blastocysts are preferred. When faced with D5 non-good-quality embryos and D6 good-quality embryos, the optimal choice was D6 4AA>D6 4BA>D6 4AB>D5 4BC>D6 4BB.

Objective:To explore the effect of the developmental stages and quality on pregnancy outcome and birth outcome, and provide evidence for single blastocyst selection in frozen-thawed cycles.Methods:A retrospective cohort study analysis was performed on the data of patients with a total of 893 cycles who underwent single blastocyst transfer in frozen-thawed cycles in the Center for Reproductive Medicine, Qingyuan People's Hospital from January 2013 to June 2021. The cycles were divided into day 5 (D5) and day 6 (D6) groups according to the time of blastocyst formation. Then the two groups were divided into four subgroups according to the quality of blastocyst, namely, D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup. The general data, clinical outcomes and neonatal outcomes of each group were compared.Results:1) The clinical pregnancy rate [60.14% (332/552)], the implantation rate [60.14% (332/552)] and the live birth rate [47.64% (263/552)] in D5 group were significantly higher than those in D6 group [45.75% (156/341), 45.75% (156/341), 36.36% (124/341), all P<0.001], but there were no significant differences in body mass index, duration of infertility, intimal thickness of transplantation day and miscarriage rate between the two groups (all P>0.05). In addition, there were also no significant differences in birth weight, low birth weight rate, fetal macrosomia rate and male/female ratio (all P>0.05). 2) There were significant differences in clinical pregnancy rate [61.00% (294/482), 54.29% (38/70), 51.00% (127/249), 31.52% (29/92)] and live birth rate [48.96% (236/482), 38.57% (27/70), 41.37% (103/249), 22.83% (21/92)] among D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup (all P<0.001). D5 good-quality embryo subgroup had the highest clinical pregnancy rate and live birth rate, while D6 non-good-quality embryo subgroup had the lowest clinical pregnancy rate and live birth rate. There were also no significant differences in birth weight, fetal macrosomia rate and male/female ratio among the four subgroups (all P>0.05), while there was a significant difference in low birth weight rate [5.08% (12/236), 0 (0/27), 4.85% (5/103), 23.81% (5/21)] among the four subgroups ( P=0.014). 3) There were no significant differences in clinical pregnancy rate and live birth rate between D5 non-good-quality embryo subgroup and D6 good-quality embryo subgroup (all P>0.05). The clinical pregnancy rate and the live birth rate of 4BC in D5 were lower than those of 4AA, 4AB and 4BA in D6, while the miscarriage rate of 4BC in D5 was higher than that of 4AA, 4AB and 4BA in D6, but there were no significant differences (all P>0.05).The clinical pregnancy rate and the live birth rate of 4BC in D5 were higher than those of 4BB in D6, but there were no significant differences (all P>0.05). Conclusion:In the frozen-thawed cycle of single blastocyst transplantation, D5 good-quality blastocysts are preferred. When faced with D5 non-good-quality embryos and D6 good-quality embryos, the optimal choice was D6 4AA>D6 4BA>D6 4AB>D5 4BC>D6 4BB.

The clinical data of 120 patients admitted to our hospital with renal stones treated by flexible ureteroscopy (FURS) and the imaging-related data measured by virtual reality technology were retrospectively analysed. The results of the univariate analysis showed that stone surface area (S), renal pelvis volume (P), length of calyces funnel (L), pelvic calyceal height (H) and essence of stone (E) were closely related to stone-free rate. The H. L.P.E.S. score was constructed to predict stone-free rate after FURS based on the above factors, and the area under the receiver aperating characteristic curve for the H. L.P.E.S. and S. O.L.V.E. scoring systems was 0.921 and 0.754 respectively.The H. L.P.E.S. scoring system has higher predictive value.

Objective To establish S.O.L.V.E.nephrolithometry scoring system,and to evaluate value of S.O.L.V.E.scoring system for predicting the stone-free rate (SFR) of flexible ureteroscopy (FURS).Methods Five reproducible variables were included in S.O.L.V.E.scoring system,such as stone surface area (S),obstruction (O),length of calyces funnel (L),visible number of calyces (V) and essence of stone (E).Variables were measured based on preoperative non-contrast computed tomography of urography.Clinical data of 392 patients who underwent FURS for upper urinary tract stones in our department from January,2017 to Jnne,2018 were retrospectively analyzed.The total study population consisted of 258 male and 134 female patients.The mean age was (49.5 ± 12.6) years old,ranged from 15 to 85 years.There were 292 patients in stone-fiee group,including 197 male and 95 female patients.The average age was (49.2 ± 12.8) years old.37 patients had previous history of renal stone surgery.Median body mass index was 24.7 kg/m2 (18.1-29.0 kg/m2) and median value of preoperative serum creatinine was 72.5 μmol/L (48.9-84.8 μmol/L).The number of patients,whose stone located in the left side and right side were 155 and 137,respectively.The number of patients,whose stones located in ureter,renal pelvis,lower calyceal and non-lower calyceal were 19,16,87,170,respectively.There were 100 patients in non-stone free group,61 men and 39 women.The average age was (50.4 ± 12.0) years old.15 patients had previous history of renal stone surgery.Median body mass index was 25.0 kg/m2 (18.5-28.8 kg/m2) and median value of preoperative serum creatinine was 73.8 μmol/L (46.5-92.5 μnol/L).The number of patients,whose stone located in the left side and right side were 51 and 49,respectively.The number of patients,whose stones located in ureter,renal pelvis,lower calyceal and non-lower calyceal were 7,4,27,62,respectively.The correlation of S.O.L.V.E.scoring system and stone-free rate,postoperative hospital stay,surgical complications,operation time were analyzed.Receiver operating characteristic curves were drawn to detect predictive value of S.O.L.V.E.scoring system for SFR of FURS.Results All cases FURS were performed successfully and the SFR was 74.5％ (292/392).Among the variables of the S.O.L.V.E.scoring system in the stone-free group and the non-stone free group,item S were (82.6 ± 69.8) mm2 and (172.6±133.7)mm2,respectively.The item L were (12.7 ± 15.8) mm and (23.9 ± 15.3)mm,respectively.The item V were (0.6 ± 0.7) and (1.3 ± 0.8),respectively.The item E were (817.1 ± 285.5) HU and (902.4 ± 256.1) HU,respectively.The difference was statistically significant (P ＜ 0.01).The item O was (17.7 ± 10.9) mm and (19.3 ± 13.1) mm,respectively,no statistical significance was found (P =0.242).The mean score was 6.3 (ranging 4-11) in this c ohort.The patients were divided into low score (4-5) group,moderate score (6-8) group and high score (9-11) group due to S.O.L.V.E.scoring system,and the stone-free rates were 93.5％ (130/139),70.5％ (153/217) and 37.5％ (9/36),respectively (P ＜0.01).The operation time of low,moderate,andi high score group were (31.6 ± 10.9),(42.3 ± 18.3),and (58.0 ± 19.2) min,respectively.Additionally,the score was correlated with the operation time(P ＜0.01),but not with postoperative hospital stay (P =0.133),intraoperative bleeding (P =0.185) and postoperative infectious fever (P =0.839).In logistic regression model analysis,the stone surface area,length of calyces funnel,number of involved calyces were significantly correlated with SFR (P ＜ 0.01).The obstruction degree and essence of stone were not associated with SFR (P ＞ 0.05).The area under receiver operating characteristic curve of S.O.L.V.E.score was 0.782,higher than that of each variable in S.O.L.V.E.scoring system(S,O,L,V,E were 0.738,0.535,0.698,0.735,0.593,respectively).Conclusions The stone surface area,length of calyces funnel,number of involved calyces were significantly correlated with SFR.The S.O.L.V.E.nephrolithometry scoring system can predict SFR after FURS accurately,and provide assistance for making clinical decisions.

OBJECTIVE:To observe therapeutic efficacy and safety of glutamine combined with low molecular weight heparin in the treatment of elderly severe pneumonia (SP).METHODS:In retrospective study,88 SP elderly patients were divided into control group (44 cases) and observation group (44 cases).Based on conventional treatment,control group was given Enteral nutritional suspension via nasal feeding.Observation group was additionally given Alanyl glutamine injection 20 g,dissolved in Amino acid solution with ratio of 1:5 at least and maximum concentration belowing 3.5％,i.v.,once a day+Low molecular weight heparin calcium injection 4 KU/kg subcutaneously,once a day,on the basis of control group.Both groups were treated for 7 d.Clinical efficacies and postoperative recovery (ventilator support time,pulmonary rale disappearance time and ICU hospitalization time) were observed in 2 groups.The levels of IgA,IgG,TNF-a,hs-CRP,APACHE Ⅱ score,coagulation function indexes (APTT,D-D,FIB,PT,PLT),blood gas analysis indexes [pa (CO2),SpO2,Oi,pa (O2)] and receptor for advanced glycation end products (RAGE),the occurrence of ADR before and after treatment were also observed in 2 groups.RESULTS:Total response rate of observation group (90.91％) was significantly higher than that of control group (72.73％),while ventilator support time,pulmonary rale disappearance time and ICU hospitalization time were significantly shorter than control group;the incidence of ADR was significantly lower than control group,with statistical significance (P＜ 0.05).After treatment,the levels of IgA,IgG,Oi,SpO2 and pa (O2) in 2 groups were significantly higher than before treatment,and the observation group was significantly higher than the control group.The levels of TNF-αt,hs-CRP,APACHE Ⅱ score,coagulation function indexes,pa(CO2) and RAGE in 2 groups were significantly lower than before,and the observation group was significantly lower than the control group,with statistical significance (P＜0.05).CONCLUSIONS:Based on conventional treatment,glutamine combined with low molecular weight heparin show significantly effect on elderly SP,effectively improve cellular immune function,coagulation function and blood gas analysis indexes and reduce RAGE level with good safety.

Objective To investigate the best pathway of laparoscopic surgery for choledocholithiasis. Methods According to MRCP,the cystic duct was divided into four types.The thick-straight type (n =89)and the thin-crooked type (n =65)patients randomly received the laparoscopic operations via the cystic duct or com-mon bile duct.The operation time,transit operation rate,complication rate and length of hospital stay after opera-tion were analysed.Results There were 46 cases of the thick-straight type via cystic duct,43 cases via common bile duct.The former had shorter operation time and hospital stay after operation (P <0.05),but there was no significant difference in transit operation rate and complication rate.There were 32 cases of the thin-crooked type via cystic duct and 33 cases via common bile duct and there were significant differences in the operation time, transit operation rate,complication rate and length of hospital stay after operation(P <0.05).Conclusion Ac-cording to different types,LCTBDE is suitable for the patients with thick-straight type,and LCHTD can be per-formed in thin-crooked type.Therefore,it can maximize the advantage of minimally invasive surgery.

MRCP can be applied to display three-dimensional morphology of the biliary tract clearly.Laparoscopic surgery via cystic duct for choledocholithiasis possesses obvious advantages, but it is not suitable for all cases because there are great variations in the cystic duct.So how to make proper choice of the therapeutic strategy becomes very important.In this prospective study, different surgeries were administrated in patients with common bile duct stone based on the 4 categories of cystic ducts in 96 cases judged by MRCP images.These findings demonstrated that according to the cystic duct morphological type,LCTBDE is suitable for thick-straight type, and LCBDE is fit for thin-crooked type.Be careful to choose laparoscopic surgery in the other two types considering their individual features.Thus, this classification will be propitious to maximize the advantage of laparoscopic operation as a minimally invasive treatment.

Objective To evaluate three different Chlamydophila pneumoniae recombinant antigens for use in Chlamydophila pneumoniae serodiagnosis. Methods The recombinant plasmids pGEX6p-2/ Cpn0146,Cpn0147 and Cpn0308 were constructed and expressed as GST fusion proteins. The immunogenicity and the immunocompetence of these recombinant protein were analyzed by Western-blot and indirect ELISA. A total of 183 sera samples of patients with respiratory tract infection and 32 sera samples of patients with Chlamydia trachomatis infection were detected with indirect ELISA coated microwell plates with the purified recombinant proteins comparing with SeroCP-TM IgG ELISA kits. The positive recognition rate, sensitivity and specificity of each method were analyzed. Results GST-Cpn0146, Cpn0147 and Cpn0308 were obtained after expression and purification. The titers of the specific IgG antibodies against Cpn0146, Cpn0147 and Cpn0308 were higher than 1:6 400, 1:128 00 and 1:128 00, respectively. When the indirect ELISA was developed to detect the IgG antibody against Chlamydophila pneumoniae in 183 samples, the concordance rate between the indirect ELISA test and SeroCP-TM IgG ELISA kits were 92. 3% (Cpn0146) , 94.5% (Cpn0147) and 96.7% (Cpn0308), respectively. The recombinant Cpn0146, Cpn0147 and Cpn0308 were recognized by 71 (38.8% positive recognition rate), 75 (40.9%), and 82 (44.8%) samples, respectively. The recombinant antigen-based detection assays displayed > 97% of detection specificity and>87%of sensitivity.Condusion GST-Cpn0308 shows a better sensitivity and specificity,which suggests it could be used for developing serodiagnosis kits of Chlamydophila pneumoniae infection.