Objective To investigate the association of the major histocompatibility complex class Ⅰ chain-related antigens A (MICA)-129 gene polymorphism and soluble MICA (sMICA) levels with ulcerative colitis (UC) in Hubei Han nationality. Methods The genetic polymorphism of MICA-129 was examined using a polymerase chain reaction-sequence based test (PCR-SBT) in 256 UC patients and 460 healthy controls. From the above subjects, 80 patients and 90 healthy individuals were randomly selected for determining serum sMICA concentrations by ELISA. Results The frequencies of variant allele (G) and genotype (GG) in MICA-129 gene were significantly higher in the UC patients than in the controls(76. 8%vs 72. 2%, P =0. 060; 55.9% vs 46. 3% ,P =0. 016). Serum sMICA levels were significantly elevated in the patients compared to the controls[(576. 47 ±279. 02) ng/L vs( 182. 17 ±73. 11 ) ng/L,P ＜0. 001]. In addition, the sMICA levels were higher in the patients carrying MICA-129 GG genotypes than in those carrying ( GA + AA) genotypes [( 638. 87 ± 347. 15 ) ng/L vs ( 507. 51 ± 152. 87 ) ng/L, P = 0. 035].Conclusions The genetic polymorphism of MICA-129 and sMICA levels are correlated with the UC patients in Hubei Han nationality. Our findings demonstrate that MICA-129 gene may contribute to the pathogenesis of UC.

Objective To explore the experiences of the diagnosis and treatment of urothelial tumor in multiple organs.Methods Clinical data of 10 patients with urothelial tumor in multiple organs were retrospectively reviewed.Urothelial tumors were found in two or more organs at the same time by B ultrasound,IVU,R-P,CTU,MRU,cystoscopy,ureteroscopy and so on before operation.Results 6 cases were operated by radical total nephroureterectomy and partial cystectomy,3 cases were operated by radical total nephroureterectomy and cystectomy with urinary diversion,1 case was operated by partial ureterectomy and total cystectomy.8 of them were alive,1 case was operated by total urethrectomy because of tumor recurrence in the posterior urethra,one died of metastasis tumor 18 months after operation,and the other died 32 month after operation.Conclusions Combined use of various kinds of the diagnostic means (ultrasound,IVU,R-P,CTU,MRU,cystoscopy,ureteroscopy) are important for the diagnosis of urothelial tumor in multiple organs.It needs to select the operate mode according to the tumor staging and grade and the patient's condition.Reinforcement surveillance and close follow up is required after operation.

Objective To evaluate association of plasma levels of homocysteine, folate and vitamin B12 as well as genetic polymorphisms of homocysteine with ulcerative colitis (UC). Methods Three hundred and ten consecutive patients with UC and 936 healthy controls were recruited.Polymorphisms of methylenetetrahyrdofolate reductase (MTHFR, C677T and A1298C), methionine synthase (MTR) A2756G and methionine synthase reductase (MTRR) A66G were genotyped using PCR-RELP methods. Eighty eight patients and one hundred healthy controls were randomly selected for determination of plasma levels of homocysteine by enzymatic cycling assay, and concentrations of folate and vitamin B12 were measured by corpuscle immune chemiluminescence assay. Results The variant allele and genotype frequencies of MTHFR 1298C, MTR 2756G and MTRR 66G were significantly higher in UC patients than in the healthy controls (P＜0. 01). Moreover, plasma homocysteine level was obviously higher in UC patients than in controls [(21.73±6.59) mmol/L vs(12.47±5.01)mmol/L,P<0.01).Whereas both folate F(11.25±6.19)nmol/L] and vitamin B12 [(322.81±128.47)pmol/L] concentrations were significantly lower in UC patients than in controls [(15.28±7.72)nmol/L and (422.59±129.36)pmol/L,respectively,P<0.01].Logistic analysis revealed that abnormal levels of homocysteine,folate and vitamin B12 were independent risk factors for UC(P<0.01).Conclusions Plasma levels of homocysteine,folate and vitamin B12 as well as the related genetic polymorphisms of homocystein are correlated with UC,which provides a theoretical basis for supplement of folate and vitamin B12 in treatment of UC patients.

ObjectiveTo review the major complications in patients after transurethal electrovaporization of the prostate (TUVP) and transurethal plasmakinetic resection of the prostate (PKRP) retrospectively and to analyze the causes and management.MethodsClinical data of 92 cases of patients after TUVP and 226 cases after PKRP were reviewed retrospectively.The patients' relevant circumstances including subjective symptoms,objective indexes and the major long-term complications were followed up about 1-,3-,and 5-year after operation.Different therapeutic methods were chosen according to different causes of the complications.ResultsThere were no significant differences (P ＞ 0.05 ) between TUVP group and PKRP group in IPSS (7.3±2.8,7.2±2.5),QOL (2.6±0.7,2.7 ±0.5),Qmax[ (25.2±3.5),(25.5 ±3.8) ml/s] and PVR [(18.7 ±5.4),(17.8 ±6.3)ml].The incidences of bladder neck restriction was about 1.1％,3.3％,and 2.3％ after 1,3,and5 years in patients after TUVP,and 0.9％,2.7％,and 1.8％ after PKRP accordingly.For urethral stricture,it was about 3.3％,2.2％,and 1.1％ after TUVP,and 3.1％,2.2％,and 0.9％ after PKRP.For residual prostatic hyperplasia,it was about 1.1％,2.2％,and 4.5％ after TUVP,and 1.3％,2.7％,and 3.7％ after PKRP accordingly.ConclusionsTUVP and PKRP are effective and safe treatment options for BPH.The major long-term complications after TUVP and PKRP are bladder neck restriction,urethral stricture and residual prostatic hyperplasia.Regular and long-term follow-up is required for patients after TUVP and PKRP.Different therapeutic methods should be chosen according to different causes of the complications after operation.

Objective To evaluate the efficacy and safety of solifenacin in the treatment of overactive bladder (OAB) syndrome in patients who have undergone transurethral resection of the prostate (TURP). Methods According to the Overactive Bladder Symptom Score (OABSS), 64 cases with OAB symptoms after TURP were randomly assigned into study and control groups with 32 cases in each group. Patients in the study group were treated with solifenacin (5 mg once daily) for a two week period beginning the first day after catheter removal. Patients in the control group were not treated with solifenacin. The mean urgency episodes, micturition episodes, nocturia, urge incontinence, volume voided per micturition, Qmax and OABSS scores were recorded on the 7th and the 14th day after catheter removal. Treatment-emergent adverse events with solifenacin in the study group were recorded and evaluated as well. All cases were followed-up for 8 weeks after catheter removal. Results There were statistically significant differences (P<0.01) in favor of the study group over the control group in the aspect of urgency, micturition episodes, nocturia, urge incontinence, volume voided per micturition and OABSS scores. The incidences of treatment related adverse events were 12.5% (4/32) in the study group with no serious adverse event observed. Conclusions Solifenacin is effective in the treatment of OAB syndrome after TURP and is well tolerated as well. Application of solifenacin should be recommended earlier after TURP.

This study was aimed to construct transgenic mouse model with target for Runxl gene. Runxl cDNA of mice was amplified by PCR from pcDNA3. 1 Flag Runx1 FL vector and inserted into ptetO7-Asc-IRES-EGFP vector to form a recombinant vector, and then the recombinant vector was injected into fertilized egg by microinjection technology to get a transgenic mouse. The results of PCR and Southern blot indicated that the Runx1 transgenic mouse was constructed successfully, and this could provide an important tool for studying the function of Runxl gene in vivo.
Animals ; Base Sequence ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Genetic Vectors ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; genetics ; Microinjections ; Molecular Sequence Data ; Recombination, Genetic

Objective To screen out and validate the abnormally expressed circular RNAs (circRNAs) in intrahepatic cholangiocarcinoma (iCCA) by comparing the circRNA microarray results of iCCA tissue and adjacent tissue, and to investigate their potential mechanism in iCCA. Methods Tumor tissue specimens were collected from three patients with iCCA who were admitted from July to December, 2019, and microarray hybridization was used to measure the differential expression of circRNAs between iCCA tissue and adjacent tissue. A total of 15 patients with iCCA who were treated during the same period of time were enrolled, and quantitative real-time PCR was used for validation of differentially expressed circRNAs. A bioinformatics analysis was performed to identify the downstream molecules of differentially expressed circRNAs, and quantitative real-time PCR was used for validation of potential molecules. The paired samples t -test was used for comparison of continuous data between groups, and the chi-square test or the Fisher's exact test was used for comparison of categorical data between groups. Results With 1.5-fold as the cut-off value for differential expression, there were 171 upregulated circRNAs and 104 downregulated circRNAs in iCCA tissue compared with the adjacent tissue. With 3-fold as the cut-off value for differential expression, there were 10 upregulated circRNAs (circRNA_002172, circRNA_002144, circRNA_001588, circRNA_000166, circRNA_000585, circRNA_000167, circRNA_402608, circRNA_006853, circRNA_001589, and circRNA_008882) and 3 downregulated circRNAs (circRNA_406083, circRNA_104940, and circRNA_006349) compared with the adjacent tissue. Pathologically confirmed iCCA tissue samples and adjacent tissue samples were collected from 15 patients, and quantitative real-time PCR was used for the validation of differentially expressed circRNAs; the results showed that circRNA_000585 was significantly upregulated in iCCA tissue ( t =3.607, P =0.003). Further bioinformatics analysis showed that circRNA_000585/miR-615-5p/AMOT/YAP might be the potential pathway involved in the pathogenesis of iCCA, and quantitative real-time PCR showed that for this pathway, miR-615-5p was significantly downregulated in iCCA ( t =5.724, P < 0.001) and AMOT and YAP were significantly upregulated in iCCA ( t =2.664 and 2.986, P =0.019 and 0.009 8). Conclusion Abnormal expression of various circRNAs is observed in iCCA, among which circRNA_000585 is significantly upregulated in iCCA and may play an important role in the pathogenesis of iCCA via the circRNA_000585/miR-615-5p/AMOT/YAP pathway.

Objective To explore the mechanism of Buyang Huanwu Decoction against cerebral ischemia-reperfusion injury in rats by regulating lipid metabolism through the cAMP/PKA/PPARγ pathway.Methods 60 rats were randomly divided into sham operation group(Sham),model group(Model),Buyang Huanwu Decoction low-dose group(BHD-L),Buyang Huanwu Decoction medium-dose group(BHD-M),Buyang Huanwu Decoction high-dose group(BHD-H)and Butylphthalide group(NBP).The cerebral ischemia-reperfusion model was prepared by transient middle cerebral artery embolization.The BHD low-,medium-and high-groups were given different doses of Buyang Huanwu Decoction(6.413,12.825,25.65 g·kg-1)by intragastric administration.The NBP group was administered with Butylphthalide(54 mg·kg-1).The sham operation group and the model group were administered with an equal volume of distilled water,all given for 14 days.The rats were subjected to neurobehavioral scoring.HE staining was used to observe brain pathological changes,and the kit was used to detect the levels of phosphocholine(PC),phosphatidylethanolamine(PE),diacylglycerol(DAG),and free fatty acid(FFA)on the ischemic side.RT-qPCR and Western Blot were applied to detect the mRNA and protein expressions of cyclic adenosine monophosphate(cAMP),protein kinase A(PKA),and peroxisome proliferator-activated receptor γ(PPARγ).Results Compared with the sham group,the neurological deficit score was significantly increased(P＜0.01),pathomorphological damage in ischemic cortex was found,the contents of PC and PE were reduced,the contents of DAG and FFA were increased(P＜0.01),and cAMP mRNA expression increased(P＜0.05)in the model group.Compared with the model group,the neurological deficit score of the BHD-L group was decreased(P＜0.05),and the neurological deficit score of the BHD-M,BHD-H and NBP groups was significantly decreased(P＜0.01),the cells in each treatment group were regularly arranged,the intercellular spaces were reduced,and the normal cells were increased.PC and PE were significantly increased,DAG and FFA were significantly decreased(P＜0.01)in the BHD-M,BHD-H and NBP groups.PC was increased,FFA and DAG were decreased in the BHD-L group(P＜0.05,P＜0.01).The mRNA level of PPARγ was increased in the BHD-L group(P＜0.05),and the mRNA and protein levels of cAMP,PKA,and PPARγ were increased in the other treatment groups(P＜0.05,P＜0.01).Conclusion Buyang Huanwu Decoction has a neuroprotective effect on cerebral ischemia-reperfusion injury rats,and its mechanism may be related to regulating the expression of key factors in the cAMP/PKA/PPARγ signaling pathway and lipid metabolism.

Objective To observe the effect of Tongxie Yaofang-containing serum on the cell cycle and apoptosis of colon cancer HCT116 cells,and to explore its possible biological mechanism.Methods Colon cancer HCT116 cells were divided into blank group and Tongxie Yaofang-containing serum group with different concentrations.Cell proliferation and activity detection-8(CCK8)was used to detect the effect of each group on the viability of HCT116 cells at 24,48 and 72 h;Flow cytometry was used to detect the apoptosis and cycle arrest of HCT116 cells induced by different concentrations of Tongxie Yaofang-containing serum;Western blot was used to detect the protein expression level of Cell Cycle Regulatory Protein P21(P21),Cyclin B1,cyclin-dependent protein kinases-1(CDK1),B-lymphoma-2(Bcl-2),Bcl-2-related X protein(Bax),phosphatidylinositol-3-kinase(PI3K),phosphorylated phosphatidylinositol-3-kinase(p-PI3K),protein kinase B(Akt)and phosphorylated protein kinase B(p-Akt)after different concentrations of Tongxie Yaofang-containing serum intervention.Results CCK8 showed that compared with the 10%blank group,10%Tongxie Yaofang-containing serum had a significant inhibitory effect on the viability of HCT116 cells only at 48 h(P<0.01);Compared with the 20%blank group,20%Tongxie Yaofang-containing serum could inhibit the viability of HCT116 cells at 48 and 72 h(P<0.01,P<0.05),and the increase of blank serum will not inhibit the viability of HCT116 cell,so 10%blank serum and 48 h were selected as the drug control and intervention time in the subsequent experiment.Flow Cytometry showed that,compared with the blank group,10%and 20%Tongxie Yaofang-containing serum could arrest HCT116 cell cycle in G2/M phase after 48 h of intervention(P<0.01),and induce HCT116 cell apoptosis.At the same time,Western blot showed that Tongxie Yaofang-containing serum could increase the expression of P21 and Bax to varying degrees,and reduces Cyclin B1,CDK1,Bcl-2,p-PI3K,p-Akt expression(P<0.05,P<0.01).Conclusion Tongxie Yaofang can inhibit the proliferation and induce apoptosis of colon cancer HCT116 cells,and its mechanism may be related to the inhibition of PI3K/Akt signaling pathway.

Objective To explore the effects of Buyang Huanwu Decoction on neuronal cell apoptosis after cerebral ischemia based on mediating Cav1 in regulating Wnt pathway.Methods Male wild-type(WT)and Cav1-/-(KO)C57BL/6 mice were randomly divided into sham-operation group,model group and Buyang Huanwu Decoction group(18.5 g/kg).Cerebral ischemia model was prepared using middle cerebral artery occlusion method,and drug intervention was given for 14 days.Neurobehavioral score was performed,HE staining was used to observe the morphology of ischemic cortical area of brain tissue,TUNEL staining was used to detect neuronal apoptosis in ischemic cortical area,immunohistochemistry was used to detect the expressions of apoptosis related proteins and Wnt1,glycogen synthase kinase 3β(GSK3β)and β-catenin protein in ischemic cortical area.Results Compared with the same genotype sham-operation group,the neurobehavioral score of the model group mice significantly increased,neuronal cells in the ischemic cortical area showed vacuolar changes,with nuclear condensation and widened intercellular spaces,the apoptosis rate of nerve cells significantly increased,with increased expressions of Bax,GSK3β and decreased expressions of Bcl-2,Wnt1 and β-catenin(P<0.01).Compared with the same genotype model group,the neurobehavioral score of mice in Buyang Huanwu Decoction group were significantly decreased,the pathological damage of the ischemic cortical area improved,the apoptosis rate of nerve cells decreased,the expressions of Bax and GSK3β decreased,and the expressions of Bcl-2,Wnt1 and β-catenin increased(P<0.01).Compared with the WT model group,the KO model group showed an increase in neurobehavioral score,aggravated damage in ischemic cortical area,significantly increased neuronal apoptosis rate,and increased expression of GSK3β(P<0.05).Compared with the WT Buyang Huanwu Decoction group,the KO Buyang Huanwu Decoction group showed an increase in neurobehavioral score,aggravated damage in ischemic cortical area,significantly increased neuronal apoptosis rate,increased expressions of Bax and GSK3β,and decreased expressions of Bcl-2,Wnt1 and β-catenin(P<0.01).Conclusion Buyang Huanwu Decoction can inhibit neuronal cell apoptosis after cerebral ischemia,and its mechanism may be related to regulating the expressions of apoptosis-related proteins by mediating Cav1 to regulate the Wnt signaling pathway.