Objective To observe the effects and study the underlying mechanism of siRNA targeting PARP1 on the proliferation of androgen independent prostate cancer PC3 cell line.Methods Three specific siRNA sequences targeting PARP1 were designed and synthesized.And two sequences which had better interfering effect on the expression of PARP1 were evaluated and selected through lipofectamine transfection,RT-PCR and Western Blot.The effect of PARP1 silencing on the proliferation of PC3 cells was observed with MTS assay and the levels of the phosphorylation of Akt and GSK3β were detected by Western Blot.Results Compared to the blank control group,the transfected group with the negative control sequence had no significant impact on the expression of PARP1,however the transfected group with siRNA-1706,-2003 or-2907 could significantly suppress the mRNA and protein expression of PARP1.The mRNA inhibition rate reached to(52.07 ± 4.65)％,(44.38 ± 9.15)％ and(22.05 ± 6.65)％,respectively;and the protein inhibition rate reached to(86.86 ± 4.94)％,(83.30 ± 7.18)％ and(63.05 ± 10.19)％,respectively.The siRNA-1706 and-2003 could significantly inhibit the proliferation of PC3 cells;the inhibition rate was(38.93 ± 3.87)％ and(34.93 ± 1.21)％.And they also could down-regulate the intracellular levels of phosphorylated Akt and GSK3β in PC3 cells.Conclusion PARP1-targeted siRNA can significantly suppress the expression of endogenous PARP1 and inhibit the proliferation of PC3 cells,which is related to the inhibition of Akt activity and the activation of GSK3 β.

Objective To investigate the effect and potential mechanism of autophagy inhibitor chloroquine on the calcium oxalate crystals formation in rats.Methods From September 2016 to October 2016,Thirty healthy male SD rats were randomly divided into 3 groups:control group,model group and chloroquine intervention group.The method to establish calcium oxalate stone model was drinking water with 1％ ethylene and 1％ ammonium chloride freely.The rats of chloroquine intervention group were treat with chloroquine (40mg/kg · d) by intraperitoneal injection.Modeling was finished after 28 days.The amounts of renalcalcium oxalate crystals were detected by polarizing microscope.For all groups,the amounts of autophagosome were detected by transmission electron microscope.Twenty four hour urine compositions for stone risk factors were detected.The expressions of oxidative stress injury related molecular markers (SOD,MCP-1 and 8-OHdG) and the expressions of autophagy markers (LC3 and P62) were detected by immunohistochemistry.The RNA expressions of SLC26A6 in kidney were detected by Real-time PCR.Results Compared to the model group,the amounts of renal calcium oxalate crystals were significantly reduced in chloroquine intervention group (32.37 ± 5.14 vs.4.18 ± 0.25,P ＜ 0.05).Compared to the control group,the level of autophagy was increased in the model group.Compared to the model group,the level of autophagy was inhibited in the chloroquine intervention group.For control group,model group and chloroquine intervention group,the excretion of urinary oxalate were (3.1 ± 1.5) mmol,(22.5 ± 8.1) mmol,(2.8 ± 1.2) mmol,respectively;the excretion of urinary citrate were (63.4 ± 7.4) mmol,(45.9 ± 9.5)mmol,(15.6 ± 8.2) mmol,respectively.Compared to the control group,the amounts of urinary oxalate weresignificantly elevated in model group (P ＜ 0.05),but citrate were significantly reduced in the chloroquineintervention group(P ＜ 0.05).For control group,model group and chloroquine intervention group,theexpressions of SOD were 42.24 ±4.16,19.21 ± 2.25,39.08 3.53,respectively;the expressions of MCP-1 were 4.02 0.51,8.45 ± 0.55,5.52 ± 0.34,respectively;the expressions of 8-OHdG were 7.16 ± 0.54,11.21 ± 1.12,8.67 ±0.34,respectively;the RNA expressions of SLC26A6 were 0.35 ±0.07,1.02 ±0.17,0.70 ± 0.06,respectively.Compared to the control group,the expressions of SOD were significantly reduced in the model group,but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜0.05).Compared to the model group,the expressions of SOD were significantly elevated chloroquine intervention group (P ＜ 0.05),but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜ 0.05).Conclusions The autophagy inhibitor chloroquine could inhibit the formation of calcium oxalate crystals induced by ethylene in rat kidney via inhibit the renal autophagy level and expressions of the SLC26A6,reducing the renal oxidative stress injury and urinary oxalate excretion.

Objective To evaluate the effect and mechanism of cysteine rich protein 61, namely CCN family member 1(CCN1) on the survival of adipose tissues in rats after autologous fat grafting. Methods At 1 week after the establishment of autologous fat grafting rat models, all animals were randomly divided into the CCN1 group (n=20) and control group (n=20). The survival of fat grafts, the morphology of fat graft tissues, the proportion of active adipocytes and the number of new blood vessels of rats were statistically compared between two groups. The levels of differential expressed messenger ribonucleic acid (mRNA) in the fat graft tissues of rats were compared between two groups by high-throughput sequencing and subsequently subject to cluster analysis. The expression levels of related proinflammatory cytokines of fat graft tissues of rats were statistically compared between two groups. Results The weight retention rate of adipose tissues in the CCN1 group was significantly higher than that in the control group (P < 0.05). In the CCN1 group, the integrity of adipocytes was considerably higher, the degree of vesiculation and vacuolation, the degree of inflammatory cell aggregation and the degree of fibrosis were significantly lower than those in the control group (all P < 0.000 1). Immunofluorescence staining demonstrated that the proportion of active adipocytes with uniform morphology was higher in the CCN1 group, whereas the proportion of active adipocytes was lower and the cells were observed in different sizes accompanied by vesiculation in the control group. Compared with the control group, the quantity of new blood vessels was significantly higher, and the expression levels of platelet derived growth factor (PDGF) and fibroblast growth factor (FGF) mRNA were remarkably higher in the CCN1 group (all P < 0.05). High-throughput sequencing analysis showed that the data at the transcriptome levels significantly differed between two groups. In the CCN1 group, the gene expression levels of cell surface markers, inflammatory cytokines and chemokines related to M1 macrophages tended to decline. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) revealed that the mRNA expression levels of interleukin (IL)-8, IL-1 and Toll-like receptor (TLR) 2 in the CCN1 group were significantly lower than those in the control group (P < 0.01-0.05). Conclusions During autologous fat grafting, supplement of exogenous CCN1 may effectively promote the neovascularization of adipose tissues and improve the survival rate of fat graft probably by mediating the transformation of macrophages into M2 phenotype via down-regulating the TLR2 expression level.

Objective To explore the significance of the initial clinical symptoms and clinical manifestations of neuroblastoma(NB)to achive early identification of NB. Methods A retrospective study was performed on patients diagnosed with NB,who attended the Hematology Oncology Center,Beijing Childrenˊs Hospital from March 31st ,2007 to March 31st,2015. The clinical characteristics were compared between the children 〈1_year_old and≥1_year_old. The result was analyzed to compare the difference in clinical symptoms and tumor biologic characteristics of patients with different initial clinical symptoms between 2 groups. Results A total of 330 patients were included in the study,43 of them were younger than 1 year old,and their most common symptoms were cough and fever( each 17 cases,and accounted for 39. 5%,respectively);while the most common symptoms in patients≥1_year_old(287 cases)was fe_ver(177 cases,61. 7%),followed by lymphadenopathy(107 cases,37. 3%),bone pain(97 cases,33. 8%)and anemia (48 cases,16. 7%). The frequency of symptoms differed significantly between 2 groups(all P〈0. 05),such as fever (39. 5% vs. 61. 7%,χ2 ﹦6. 68),anemia(4. 7% vs. 16. 7%,χ2 ﹦6. 00),bone pain(0 vs. 33. 8%,χ2 ﹦18. 99),abdo_minal pain(0 vs. 25. 3%,χ2 ﹦10. 19),diarrhea(16. 3% vs. 3. 0%,χ2 ﹦12. 73),lymphadenopathy(7. 0% vs. 37. 3%, χ2 ﹦14. 12)and anorexia(9. 3% vs. 33. 4%,χ2 ﹦9. 21). Datients had fever,anemia,lymphadenopathy,cutaneous hemorrhagic spot or periorbital ecchymosis,bone pain,abdominal pain,exophthalmos,and anorexia early in the initial course of the disease,whose serum lactate dehydrogenase values were significantly increased(P〈0. 05). Datients suf_fered from fever,anemia,lymphadenopathy,bone pain,limbs dysfunctions,abdominal pain at the beginning of the disease,whose urine vanillymandelic acid values were higher than normal(P〈0. 05). Conclusions The study of sympto_mology suggests the most common symptoms in patients with NB 〈1_year_old are cough and fever,while those ≥1_year_old are fever,lymphadenopathy,bone pain,and anemia. Por patients with symptoms mentioned,carrying out the necessary NB_diagnose_related laboratory and imaging studies was statistically relevant to patientsˊ ages(〈1_year_old and≥1_year_old),which may contribute to earlier identification and diagnosis of NB.

Objective To identify risk factors of systemic inflammatory response syndrome (SIRS) after percutaneous nephrolithotomy (PCNL).Methods We retrospectively reviewed 438 renal calculi patients after PCNL from August 2015 to July 2016.Among them,there were 251 men and 187 women,the mean age was (49.4 ± 11.1) years.The positive preoperative urine WBC,culture and nitrite rates were 29.7％ (130),12.1％ (53) and 15.1％ (66),respectively.The stone size was (851.2 ± 663.6) mm2,the stone CT value was (960.4 ± 303.4) HU,the operative time was (63.5 ± 33.4) min,124 (28.3 ％) were infection stones and multiple-tracts PCNL was performed in 69 (15.8％) patients.Univariate and multivariate logistic regression analysis were used to analyze perioperative predictors after PCNL.Results Thirty-nine patients developed SIRS (8.9％) after PCNL.The univariate analysis showed that positive preoperative urine WBC,nitrite,culture,operation time,stone size and transfusion had significantly impacts on the outcome of postoperative SIRS after PCNL (P ＜ 0.05).Multivariable logistic analysis showed that positive preoperative urine nitrite (OR =5.990,P ＜ 0.001),stone size (OR =2.251,P =0.027) and transfusion (OR =7.501,P =0.007) were independently related to the postoperative SIRS.Conclusion The positive preoperative urine nitrite,stone size and transfusion are independent risk factors for postoperative SIRS after PCNL.

Objective:To assess the effect of stone and urine bacteria culture on the treatment of postoperative infection in percutaneous nephrolithotomy (PCNL).Methods:Between September 2016 and September 2018, 1060 patients with kidney stones treated with first-stage PCNL were included in the study. There were 614 male and 446 female patients, with the mean age (52.4±12.2) years. The mean stone burden was (1 499.6±1 435.3) mm 2. The midstream urine sample and the stone sample were sent for bacterial culture, identification of bacterial strain and antimicrobial susceptibility tests. The results of urine culture (UC), stone culture (SC) and their antimicrobial susceptibility, the details of perioperatively administered antibiotics and postoperative infections were recorded. The relationship between the postoperative infection and the SC was analyzed. Results:In 1 060 patients, 22 bacterial species were identified in UC and 52 bacterial species were identified in SC. The positive rate was higher in SC than in UC[31.8%(337/1 060)vs. 20.9%(222/1 060), P<0.001]. Escherichia coli was the most common bacteria in both UC and SC, but was more prevalent in UC than in SC [52.3%(116/222)vs. 43.6%(147/337), P<0.05]. E. coli cultured from UC and SC had high resistance to ampicillin, cefazolin, ceftriaxone, cefotaxime, levofloxacin, ciprofloxacin (all resistance rate >40%), but were sensitive to meropenem, cefoperazone/sulbactam, piperacillin/tazobactam, and amikacin (all resistance rate <10%). There was no statistical difference in the antibiotic resistance rates of E. coli from the UC and SC (all P >0.05). There were 111 (10.5%) patients who developed fever and 22 (2.1%) who developed urosepsis postoperatively. The incidences of postoperative fever and urosepsis were higher in the patients with positive SC than the patients with negative SC [23.7%(80/337)vs. 4.3%(31/723); 4.2%(14/337)vs. 1.1%(8/723), P<0.05]. Even in patients with negative UC, The incidence of postoperative fever was higher in the group with positive SC than the group with negative SC [17.9%(30/168) vs. 4.2%(28/670), P<0.05]. The incidence of postoperative fever in SC positive patients was lower if they were treated with sensitive antibiotics to the bacteria in stone than those treated with nonsensitive antibiotics [17.5%(22/126) vs. 27.5%(58/211), P<0.05]. Conclusions:The SC had high rate of culture positive, complicated bacterial species and high rate of multi-drug resistant. Positive SC was associated with increased incidence of postoperative infection even if the patients had negative UC. The SC might have a importance clinical value in the treatment of postoperative infection in PCNL.

Objective:To investigate the efficacy of low-dose uric acid oxidase in treating children with aggressive mature B-cell non-Hodgkin lymphoma accompanied by hyperuricemia.Methods:Clinical data of children with primary aggressive mature B-cell non-Hodgkin lymphoma and hyperuricemia, who were treated in Beijing Children′s Hospital, Capital Medical University from January 2016 to June 2021 were retrospectively analyzed.The serum uric acid concentration was monitored in all pediatric patients from the day before chemotherapy to the seventh day of chemotherapy.Low-dose uric acid oxidase [0.05-0.10 mg/(kg·dose)] was intravenously injected into the patients when the serum uric acid level exceeded the upper limit of the normal range.The therapeutic effect and clinical medication experience of uric acid oxidase were summarized.The change of serum uric acid levels with time before and after the application of different doses of uric acid oxidase was analyzed by a repeated measures ANOVA. Results:A total of 106 children with primary aggressive mature B-cell non-Hodgkin lymphoma and hyperuricemia were enrolled in this study.There were 88 males and 18 females, with a median age of 6.5 (3.5, 10.0) years.The pathological subtypes comprised Burkitt′s lymphoma in 95 cases (89.6%), high-grade B-cell lymphoma in 7 cases (6.6%), and diffuse large B-cell lymphoma in 4 cases (3.8%). Additionally, 39 cases (36.8%) were in clinical stage Ⅲ and 67 cases (63.2%) were in stage Ⅳ.All cases had high tumor burden, including renal involvement in 52 cases (49.1%), tumor lysis syndrome in 42 cases (39.6%), and acute kidney injury in 27 cases (25.5%). Totally, one dose of uric acid oxidase was intravenously injected into 41 children (38.7%), 41 children (38.7%) were given 2 dosages, 20 children (18.9%) were given 3 dosages, and 4 children (3.8%) received 4 dosages.Moreover, 9 cases (8.5%) were supplemented with continuous renal replacement therapy.Serum uric acid concentrations before chemotherapy and 12 hours after injecting the first dose of uric acid oxidase were (741.4±312.9) μmol/L and (210.8±148.6) μmol/L, respectively.The difference was statistically significant ( t=5.288, P<0.001). The change of serum uric acid levels over time before and after the application of different doses of uric acid oxidase in children was compared, and no significant difference was found ( F=0.225, P=0.879). No delay in chemotherapy or death arising from tumor lysis syndrome and acute kidney injury occurred within 28 days after chemotherapy. Conclusions:Low-dose and on-demand application of uric acid oxidase can rapidly and effectively reduce serum uric acid levels in children with aggressive mature B-cell non-Hodgkin lymphoma in the early stage of chemotherapy.

To investigate the effects of ventricular shunt placement in children with post-infective hydrocephalus.
 Methods: A total of 24 cases of post-infectious hydrocephalus, who received ventricular shunt, were enrolled. Age, gender, disease progression, clinical manifestation, laboratory data, treatment, prognosis, complication, and sequela for each patient were retrospectively reviewed.
 Results: Of the 24 cases, 8 had a full recovery, 11 had slight sequela, 2 had severe sequela, 1 was in vegetative state, and 2 died because of bacterial meningitis and tubercular meningitis. Epilepsy, mental impairment, visual and hearing damage were the main sequelae.
 Conclusion: Ventricular shunt is the preferred treatment in children with post-infective hydrocephalus, which shows positive clinical efficacy and can improve the long-term prognosis of such patients.
Child ; Humans ; Hydrocephalus ; Retrospective Studies ; Treatment Outcome ; Tuberculosis, Meningeal ; Ventriculoperitoneal Shunt

Overactive bladder (OAB) is the most bothersome symptom in lower urinary tract symptoms (LUTS). Current pharmacologic treatment aims to inhibit detrusor contraction; however, shows unsatisfied efficacy and high discontinuation rate. LIM kinases (LIMKs) promote smooth muscle contraction in the prostate; however, their function in the bladder smooth muscle remains unclear. Here, we studied effects of the LIMK inhibitors on bladder smooth muscle contraction and proliferation both