Objective:To explore the changes of immune microenvironment and prognosis of bladder cancer patients with positive urinary nuclear matrix protein 22 (NMP22).Methods:Retrospective analysis was made on 86 patients who were diagnosed with bladder cancer in Xuzhou Central Hospital from January 2019 to September 2020. All patients were tested for urinary NMP22 by colloidal gold method. The patients with positive test results were NMP22 positive group, and the patients with negative test results were NMP22 negative group. The expression of CD8, programmed cell death-ligand 1 (PD-L1), programmed cell death protein-1 (PD-1) and PanCK were detected by multiple fluorescent immunohistochemical method on the pathological tissue sections of all enrolled patients with bladder cancer after surgery. Follow-up data of enrolled patients were collected after discharge, and univariate and multivariate Cox analysis was performed on the follow-up data.Results:There were 69 patients in the NMP22 positive group and 17 patients in the NMP22 negative group. The percentage of CD8 and PD-L1 positive cells in NMP22 positive group was significantly higher than that in NMP22 negative group, and the difference was statistically significant (all P<0.05). Univariate analysis showed that tumor stage was correlated with bladder cancer progression ( HR=2.67, P=0.017). Multivariate analysis showed that positive NMP22 was significantly correlated with bladder cancer recurrence and disease progression (all P<0.05). Conclusions:The density of CD8 + T cells and PD-L1 in tumor parenchyma of urinary NMP22 positive bladder cancer patients was higher than that of NMP22 negative patients. Urinary NMP22 positive can be one of the bad prognostic factors of bladder cancer, and the patients with NMP22 positive should strengthen reexamination.

Objective:To establish and verify a dynamic web-based nomogram for predicting futile recanalization after thrombectomy in acute ischemic stroke.Methods:Three hundred and four acute ischemic stroke patients admitted to the Second Affiliated Hospital of Soochow University from May 2017 to April 2021 were retrospectively enrolled. All these patients underwent mechanical thrombectomy and obtained successful recanalization. The eligible patients were randomly divided into training group ( n=216) and test group ( n=88) by 7∶3. The nomogram was established and internally validated with the data of the training group, and externally validated with the data of the test group. For the training group, multivariate Logistic regression analysis was performed by including all variables with P<0.05 in univariate analysis, and the independent predictors of futile recanalization were screened out to construct a nomogram. In the training group and the test group, the performance of the nomogram was verified by C-index, calibration chart and decision curve analysis respectively. Results:No significant difference was detected between the training group and the test group in futile recanalization [134/216 (62.0%) vs 56/88 (63.6%), χ 2=0.07, P=0.794]. Multivariate Logistic regression analysis showed that age ( OR=1.04,95% CI 1.00-1.08, P=0.033), National Institutes of Health Stroke Scale (NIHSS) score on admission ( OR=1.11,95% CI 1.04-1.19, P=0.001), neutrophil to lymphocyte ratio ( OR=1.19,95% CI 1.07-1.32, P=0.001), glycated hemoglobins ( OR=2.02,95% CI 1.34-3.05, P<0.001), poor collateral status ( OR=10.87,95% CI 4.08-29.01, P<0.001), postoperative high density ( OR=11.38,95% CI 4.56-28.40, P<0.001) were independent risk factors for futile recanalization. The C-index of this nomogram in the training group and the test group was 0.92 (95% CI 0.877-0.954, P<0.001) and 0.93 (95% CI 0.87-0.98, P<0.001), respectively. Conclusion:This web-based nomogram, including age, NIHSS score on admission, neutrophil to lymphocyte ratio, glycated hemoglobin, poor collateral status and postoperative high density, predicted individual probability of futile recanalization after mechanical thrombectomy with good discrimination and clinical utility.

Objective:To assess the efficacy and safety of 100 units of botulinum toxin A (BTX-A) intradetrusor injection in patients with overactive bladder.Methods:From April 2016 to December 2018, 17 tertiary hospitals were selected to participate in this prospective, multicenter, randomized, double-blind, placebo-controlled study. Two phases of study were conducted: the primary phase and the extended phase. This study enrolled patients aged 18 to 75 years who had been inadequately managed by anticholinergic therapy (insufficient efficacy or intolerable side effects) and had spontaneous voiding with overactive bladder. Exclusion criteria included patients with severe cardiac, renal and hepatic disorders, patients with previous botulinum toxin treatment for 6 months or allergic to BTX-A, patients with urinary tract infections, patients with urinary stones, urinary tract tumors, diabetes mellitus, and bleeding tendency. Eligible patients were randomly assigned to BTX-A group and placebo control group in a ratio of 2∶1. Two groups of patients received 20 intradetrusor injections of BTX-A 100U or placebo at the depth of the submucosal muscle layer respectively under cystoscope, including 5 injections at the base of the bladder, 3 injections to the bladder triangle, 5 injections each to the left and right walls and 2 injections to the top, sparing the bladder neck. As a placebo control group, patients received same volume of placebo containing no BTX-A and only adjuvant freeze-dried preparations for injection with the same method. A combination of gelatin, sucrose, and dextran served as adjuvants. Average micturition times per 24 hours, urinary incontinence (UI) episodes per day, average micturition volume per day, OAB symptom score(OABSS), and quality of life (QOL) score were recorded at baseline and the 2nd, 6th and 12th week after treatment. The primary efficacy endpoint was the change from baseline in the average micturition times per 24 hours at the 6th week after treatment. The secondary efficacy endpoints included the change from baseline in the average micturition times per 24 hours at 2nd and 12th week, as well as the change from baseline in the OABSS, QOL score, average frequency of urgency and UI episodes per day, urgency score, average micturition volume per day at 2nd, 6th and 12th week after treatment. Patients were followed for 12 weeks to assess adverse events (AEs). After assessed at week 12, if the micturition times has decreased less than 50% compared to baseline and the patient is willing to receive retreatment, then patients could enter the extended trial phase. In that phase, patients in both groups were injected with 100 units BTX-A from 12th week onwards and then followed up the same indicators for 12 weeks.Results:216 patients were enrolled in this trial (144 cases in the BTX-A group and 72 cases in the placebo control group). Baseline characteristics such as age (47.75±14.20 in the BTX-A group and 46.39±15.55 in the control group), sex (25 male/117 female in the BTX-A group and 10/61 in the control group), and disease duration (0.51 years in the BTX-A group and 0.60 years in the control group) were balanced between the two groups( P>0.05). A marked reduction from baseline in average micturition times per 24 hours was observed in all treatment groups at the 6th week and the reduction of the two groups was statistically different ( P<0.001 and P=0.008 respectively). Compared with the baseline, the average micturition times per 24 hours at the 6th week decreased from baseline by 2.40(0.70, 4.60)times for the BTX-A group and 0.70(-1.00, 3.30) times for the placebo control group respectively, and the difference between the two groups was considered to be statistically significant ( P=0.003). The change rates of average micturition times per 24 hours from baseline at the 6th week of the two groups were (16±22)% and (8±25)% respectively, and the difference between the two groups was statistically significant ( P=0.014). Compared with the baseline, the average micturition times per 24 hours at 2nd and 12th week decreased by 2.00(0.00, 4.00)and 3.30(0.60, 5.03)for the BTX-A group, 1.00(-1.00, 3.00)and 1.70(-1.45, 3.85)for the placebo control group respectively. The difference between two groups was considered to be statistically significant ( P=0.038 and P=0.012); the changes of average urgency times per day for the BTX-A group and the control group at the 2nd, 6th and 12th week were 2.00(0.00, 4.30)and 2.40(0.30, 5.00), 3.00(0.30, 5.70)and 0.70(-1.30, 2.70), 0.70(-1.30, 3.00) and 1.35(-1.15, 3.50), respectively. There were significant differences between two groups at the 2nd, 6th and 12th week, ( P=0.010, P=0.003 and P=0.025, respectively). The OABSS of the BTX-A group and the control group at the 6th week decreased by 1.00(0.00, 4.00)and 0.50(-1.00, 2.00) compared with the baseline, and the difference between the two groups was statistically significant ( P=0.003). 47 cases of BTX-A group and 34 cases of placebo control group entered the extended trial phase, and 40 and 28 cases completed the extended trial phase, respectively. The average micturition volume per 24 hours changed by -16.60(-41.60, -0.60)ml and -6.40(-22.40, 13.30)ml, (-35.67±54.41)ml and(-1.76±48.69)ml, (-36.14±41.51)ml and (-9.28±44.59)ml, (-35.85±43.35)ml and(-10.41±40.29)ml for two groups at the 12th, 14th, 18th and 24th week, and the difference between two groups was statistically significant at each follow-up time ( P=0.01, 0.006, 0.012 and 0.016, respectively). There was no significant difference in other parameters( P>0.05). However, adverse reactions after intradetrusor injection included increased residual urine volume (27 in the BTX-A group and 3 in the control group), dysuria (21 in the BTX-A group and 6 in the control group), urinary infection (19 in the BTX-A group and 6 in the control group), bladder neck obstruction (3 in the BTX-A group and 0 in the control group), hematuria (3 in the BTX-A group and 1 in the control group), elevated alanine aminotransferase (3 in the BTX-A group and 0 in the control group), etc. During the follow-up period, there was no significant difference in the other adverse events between two groups except the increase of residual urine volume( P<0.05). In the primary trial phase, among the 27 cases with increased residual urine volume in BTA group, only 1 case (3.70%) with PVR more than 300 ml; the PVR of 3 patients in the placebo group was less than 100 ml. The increase of residual urine volume caused by the injection could be improved or disappeared with the passage of time. Conclusions:Intradetrusor injection of Chinese BTX-A improved the average micturition times per 24 hours, the average daily urgent micturition times, OABSS, and average micturition volume per time, and reduced the adverse effects in patients with overactive bladder.Chinese BTX-A at dose of 100U demonstrated durable efficacy and safety in the management of overactive bladder.

Objective:To compare the proportion of abnormal renal function indexes in outpatients with chronic hepatitis B (CHB), and to further explore the correlation between the history of nucleos(t)ide analogues (NA) antiviral therapy and abnormal renal function indexes.Methods:A cross-sectional design was adopted for renal function screening. Baseline characteristics, history of antiviral treatment, and renal function indexes were collected, including glomerular filtration rate (eGFR), blood urea nitrogen, blood creatinine, blood uric acid, and urine β2- microglobulin α 1-microglobulin and urinary protein. According to the definition and standard of abnormal renal function indexes, the correlation between chronic kidney disease (CKD) - 1/2 and related risk factors, renal tubular indicators and risk factors, and the correlation between antiviral treatment duration and renal tubular risk were analyzed. The data were analyzed by single and multi-factor regression.Results:2703 outpatients from 47 hospitals across the country were enrolled. 70.7% were males with an average age of 47.5 years. 15.5% of cases had hypertension, 14.6% cases had chronic kidney disease, 11.3% cases had diabetes, and 15.4% had CKD 1/2. Retinol-binding protein, urinary β2-microglobulin or urinary α1-microglobulin showed renal tubular damage rates of 10.4%, 27.1% and 18.4%, respectively. Renal tubular damage risk was related to the antiviral treatment duration ( OR = 1.06, 95% CI = 1.028 ~ 1.093). Multivariate analysis results showed that the risk factors related to eGFR < 90 ml/min in male were 40-60 or > 60 years old, high viral load, poorly controlled hypertension, use of nephrotoxic drugs, liver fibrosis status, family history of hepatitis B; while the risk factors related to eGFR < 60 ml/min in female were decompensated cirrhosis, poorly controlled diabetes, and family history of hepatitis B. Conclusion:CHB outpatients have a high proportion of chronic kidney damage, including early renal tubular damage, which suggests that baseline renal function should be assessed before NA use and closely monitored during the treatment. Regular monitoring of the renal tubular damage index can detect the risk of kidney damage earlier than the estimated eGFR.

OBJECTIVE To investigate the protective effects and mechanism of diterpene ginkgolides meglumine injection (DGMI) against oxidative stress induced by oxygen-glucose deprivation (OGD) in SH-SY5Y cells. METHODS SH-SY5Y cells were divided into five groups: normal control, model control (OGD group) and drug(25 mg · L- 1) administration groups including DGMI group, extract of ginkgo biloba leaves injection group (EGBLI) and lactones ginkgo biloba injection group (LGBI). The cells suffered from oxygen-glucose deprivation (OGD) for 4 h, followed by reoxygenation with drugs for 6 h. Then, cell viabilities were detect using CCK-8 assays, reactive oxygen species (ROS) levels using fluorescence probe DCFH-DA and superoxide dismutase (SOD) activities using WST-1 test. Western blotting was used to detected protein levels of hemeoxygenase-1(HO-1), NAD(P)H, quinone oxidore?ductase l (Nqo1), protein kinase B (Akt), phosphorylated Akt (p-Akt), nuclear factor-E2-related factor2 (Nrf2) and phosphorylated Nrf2 (p-Nrf2). The cells were induced by OGD for 4 h, followed by reoxygen?ation and DGMI for 1 h, combined with different concentrations of PI3K inhibitor (LY294002) (at the final concentration of 12.5, 25 and 50 μmol · L-1) before the protein levels of AKT, p-AKT, Nrf2 and p-Nrf2 were detected by Western blotting. RESULTS SH-SY5Y cells induced by OGD for 4 h resulted in an increase in ROS(P<0.01), but a decrease in cell viabilities(P<0.01), SOD activities(P<0.01), and antioxidant protein levels ( Akt, p-Akt, Nrf2, p-Nrf2, HO-1 and Nqo1) (P<0.01). Compared with OGD group, treatment with reoxygenation and drugs (DGMI,EGBLI and LGBI respectively) for 6 h resulted in a decrease in ROS (P<0.01), but an increase in cell viabilities, SOD activities and antioxidant protein levels of p-Nrf2, HO-1, Nqo1 and p-Akt(P<0.05,P<0.01). DGMI group showed the best efficiently. Moreover, after OGD for 4 h, compared with DGMI group, combining reoxygenation and DGMI with LY294002 for 1 h resulted in a concentration-dependent inhibition of the protein levels of p-AKT and p-Nrf2(P<0.01). CONCLUSION DGMI 25 mg · L-1 can inhibit oxidative stress in SH-SY5Y cells induced by OGD by increasing the activity and expression of Nrf2 through PI3K/Akt pathway, which may be one of the mechanisms by which DGMI protects neurons from stroke.

Aim To investigate the protective effects of Diterpene Ginkgolides Meglumine Injection(DGMI)on SY5 Y cells damaged by oxygen-glucose deprivation and its functional mechanisms.Methods After 4 h of OGD,the cells were treated with 25 mg·L-1 drugs for 1 h.Subsequently,cell viabilities were measured by cell counting kit-8(CCK-8 kit)and cell apoptosis was measured by flow cytometric analysis.Furthermore, the mitochondrial membrane potential was detected by rhodamine123 staining.The levels of phospho-p38, phospho-p53,Bcl-2,Bax and cleaved caspase-9/3 were evaluated by western blot.Results DGMI signif-icantly increased the cell viabilities of SY5 Y cells dam-aged by OGD,and reduced OGD-elicited dissipation of mitochondrial membrane potential and cell apoptosis. Furthermore,DGMI also reduced p-p38,p-p53,Bax/Bcl-2 ratio,cleaved caspase-9 and cleaved caspase-3. Conclusion DGMI shows good neuroprotective effects on SY5 Y cells after oxygen-glucose deprivation.The underlying mechanisms may be associated with the sup-pression of p38/p53/Bcl-2 /caspase-9/caspase-3 sig-naling pathway.

Background and purpose:More and more evidence has showed that Grb2 binding protein-2 (Gab2) is associated with tumor invasion and metastasis. However, the relationship between Gab2 and epithelial-mesenchymal transition (EMT) in breast cancer is not clear. The aim of this study is to investigate the effect of Gab2 on EMT markers and the mechanism of Gab2 on breast cancer invasion and metastasis.Methods:Immunohistochemical methods were used to detect the expressions of Gab2, E-cadherin and vimentin in 80 cases of breast cancer tissues, and the correlations between them were analyzed. Western blot was used to detect the expression of Gab2 in breast tissues. After MDA-MB-231 cells were transfected with siRNA plasmid, wound healing assay was used to detect the invasive ability of transfected cells induced by epithelial growth factor (EGF) in vitro. Then Western blot was used to analyze the protein expressions of E-cadherin, vimentin, phosphorylated GSK-3β (p-GSK-3β) and nuclear Snail.Results:Gab2 was negatively correlated with the expression of E-cadherin and positively correlated with the expression of vimentin in breast cancer tissues (P<0.05). The expression of Gab2 in breast cancer tissues was higher than that in normal breast tissues adjacent to breast cancer. In vitro, Gab2 expression was significantly knocked down in MDA-MB-231 cells transfected with Gab2 siRNA plasmid (SiGab2/MDA-MB-231cells). Meanwhile, the invasive ability of SiGab2/MDA-MB-231cells was decreased with EGF stimulation. The expression of E-cadherin was increased in SiGab2/MDA-MB-231cells. However, the expressions of vimentin, p-GSK-3β and nuclear Snail were decreased in SiGab2/MDA-MB-231cells.Conclusion:Gab2 can promote the invasion and metastasis of breast cancer by EMT through GSK-3β/Snail signaling pathway.

Objective:To observe the effect of hypocaloric parenteral nutrition (PN) regimen on the postoperative recovery in aged patients with gastric cancer .Methods:Sixty aged patients with gastric cancer were divided into observation group (n=30) and control group (n=30) on the basis of postoperative PN feeding strategies .The observation group received hypocaloric PN which contained calorie 18 kcal/kg/d and nitrogen 0 .1 g/kg/d .And the control group received calorie 30 kcal/kg/d and nitrogen 0 .2 g/kg/d .Both groups have been provided with PN for 6 days continuously .The differences regarding C‐reactive protein(CRP) ,serum albumin ,blood glucose ,infectious complications and duration of hospital stay were compared between the two groups .Results:CRP level on the 6th day in observation group was significantly lower than that in control group , while levels of serum prealbumin and transferrin in observation group were higher than those in control group (P< 0 .05) . There was no significant difference regarding blood glucose concentration between the groups , but the dose of insulin administered on the 6th day in observation group was significantly lower than that in control group (P< 0 .05) .And both infectious complications and duration of hospital stay in observation group were fewer than those in control group (P<0 .05) . Conclusions :Hypocaloric PN regimen was conducive to postoperative recovery in aged patients with gastric cancer .

Objective The purpose of this study was to establish a rat model of thromboembolism for the study of hemorrhag -ic transformation after intravenous thrombolysis with the recombinant tissue plasminogen activator ( rtPA) . Methods Sixty Sprague-Dawley rats were randomly divided into a sham operation , a cerebral embolism, and an rtPA group.Thrombus was prepared in vitro with the rat femoral artery blood and injected into the internal carotid artery of the rats in the cerebral embolism and rtPA groups to es -tablish a model of embolic focal cerebral ischemia , while the animals of the sham operation group injected with BSA .Five hours later , the rats in the rtPA group received rtPA and those in the cerebral embol-ism and sham operation groups the injection of isotonic saline solu-tion.At 24 hours after embolus injection , the neurological deficit score was obtained .The rats were sacrificed after cardiac perfusion and their brains removed for triphenyltetrazolium chloride staining , assessment of the infarct volume and cerebral edema , and calculation of the hemorrhage volume by spectrophotometric hemoglobin assay . Results The hemorrhage volume was significantly higher in the rtPA than in the cerebral embolism group ([17.55 ±2.20] μL vs [3.82 ±0.86] μL, P<0.01), but there were no statistically significant differences between the two groups in the infarct volume ([29.29 ±4.204] %vs [27.89 ±3.91] %, P=0.810), cerebral edema ([12.43 ±1.66] % vs [7.13 ±2.04] %,P=0.063 2), and neurological deficit score (3.35 ±0.27 vs 2.80 ±0.28, P=0.174). Conclusion The rat model of thromboembolism, with a high stability and reproducibility , can be used for the pathogenesis-related studies of hemorrhagic transformation after thromboly-sis with rtPA.