Helicobacter pylori(Hp)infection is the cause of a variety of gastrointestinal disease. Currently,there are many therapeutic regimens for Hp eradication,however,because of the increase of Hp resistance and adverse effects in the treatment,the eradication rate of Hp is not very satisfactory. Studies showed that probiotics as an adjuvant may improve the eradication rate and reduce the incidence of adverse effects in eradication therapy for Hp. This article reviewed the advances in study on application of probiotics in eradication of Hp.

BACKGROUND:Because of convenient source, multi-lineage differentiation and low immunogenicity, bone marrow mesenchymal stem cels are the ideal cel type to serve as vectors of transgenic cels in pain management. However, the replicative senescence and smal amount of cels obtained from the bone marrow restrict the application of bone marrow mesenchymal stem cels in pain research. OBJECTIVE:To construct human telomerase reverse transcriptase (hTERT)-immortalized rat bone marrow mesenchymal stem cels as transgenic celular vectors for pain therapy. METHODS:Bone marrow mesenchymal stem cels were obtained from whole rat bone marrow, and then transfected with a lentivirus containing the hTERT (pLV-Puro-EF1α-hTERT) folowed by puromycin selection. hTERT expression and telomerase activity in these transfected cels were determined by RT-PCR and TRAP. Morphological changes, capacity of cel growth and multi-lineage differentiation, chromosome karyotype and tumorigenicity were observed in vitro. Moreover, the expression of cel surface molecule, Nestin, MHC-I and MHC-II in transfected cels were also detected by flow cytometry and immunocytochemistry. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cels geneticaly modified by hTERT could be cultured and passaged through 30 generations in vitro. Compared to the primary and negative transfected cels, the hTERT-modified bone marrow mesenchymal stem cels showed higher expression of hTERT mRNA, telomerase activity and cel proliferation. Most of transfected cels stayed at G2/M and S stages. The proliferation index of the transfected cels were increased dramaticaly. The positive rates of CD29, CD44 and CD90 were over 70%, but the positive rates of CD34 and CD45 were less than 5%. Transfected cels were positive for Nestin in the cytoplasm, but negative for MHC-1 and MHC-11. In addition, this cel line continued to exhibit the characteristics of fibroblastic bone marrow mesenchymal stem cels, including phenotype, differentiation into osteoblasts, adipocytes and neuron-like cels. No chromosome abnormality and tumor formation were observed in this experiment. Taken together, these data suggests that the rat bone marrow mesenchymal stem cels immortalized by hTERT gene are constructed successfuly and stil maintain major stem cels characteristics, which provide safe and stable cel vectors as research base for pain therapy.

Objective: To observe the effects of recombinant adenovirus TRAIL (AdS-TRAIL & Ad5F35-TRAIL) on apoptosis of non-small cell lung (NSCLC) cells, so as to assess the value of Ad-TRAIL in gene therapy of NSCLC. Meth-ods: CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells from samples of 10 NSCLC patients were assayed by flow cytometry analysis. The lung cancer cell lines and primary lung cancer cells were infected with Ad5-TRAIL & Ad5F35-TRAIL adenoviral vectors at MOI 10 or 50, re-spectively; the percentage of apoptosis cells labeled by Annexin V-FITC in different cells were measured by flow cytometry 48 h after transfection. Results: The expression of CD46 were higher than that of CAR in all the lung cancer lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5-TRAIL or Ad5F35-TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)% (Ad5-TRAIL), (15.96±2.89) % (Ad5F35-THAIL) and (6.05±1.58) % (Ad5-TRAIL), (10.11±1.26) % (Ad5F35-TRAIL), respectively, compared to no adenovirus-transfected cells ([2.33±0.37] % and [5.95±1.89]%, respectively, P < 0.05). Less than 10% of apoptosis cells were detected in NCI-H520 cells transfected with Ad5- or Ad5F35-TRAIL at MOI 50 ([12.89±3.2] % for AdS-TRAIL and [9.08±1.35]% for Ad5F35-TRAIL, respectively) compared to no adenovirus-transfected cells ([7.04±2.17] %, P > 0.05). Moreover, apoptosis induced by Ad5- or Ad5F35-TRAIL transfection in A549 cells was not detected both at MOI 10 and 50. About half of the primary lung cancer cells from 10 patients induced apoptosis after transfected with Ad5-TRAIL or Ad5F35-TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35-TRAIL group than those in Ad5-TRAIL and control groups. Conclusion: Ad5-TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35-TRAIL is more potent than Ad5-TRAIL, so Ad5F35-TRAIL is more suitable for gene therapy of NSCLC.

AIM: To explore the molecular mechanisms of SC58125 on apoptosis in HepG2 cells. METHODS: Cell culture, ELISA, flow cytometry, RT-PCR, Western blot and electrophoretic mobility shift assay (EMSA) analysis were employed to clarify the effect of SC58125 on apoptosis in HepG2 cells and related molecular mechanisms. RESULTS: SC58125 induced apoptosis in HepG2 cells in a concentration-dependent manner, which was accompanied by inhibition of NF-?B, activation of caspase-3, decrease of bcl-2 mRNA and increase of p53 mRNA. However, no significant changes were found in the DNA binding of AP-1. CONCLUSION: SC58125 induces apoptosis in HepG2 cells, which may be related to the inhibition of NF-?B, activation of caspase-3, decrease of bcl-2 mRNA and increase of p53 mRNA.

AIM: To clarify the effect of SC58125 on cell proliferation and apoptosis in HepG-2 cells and explore the molecular mechanisms. METHODS: Cell culture, MTT, TUNEL, DNA ladder, flow cytometry and Western blot analysis were employed in the present study. RESULTS: SC58125 inhibited the growth of HepG-2 cells and induced the apoptosis. Furthermore, it arrested G_0/G_1 phase and inhibited S phase in HepG-2 cells. Depressed expression of P33 cdk2 ,P34 cdc2 ,cyclin B_1,cyclin E ,Mpm-2,Rb ,PCNA proteins were found in HepG-2 cells treated with SC58125. CONCLUSION: SC58125 inhibits cell proliferation and induces apoptosis, which may be related to the altered low protein levels of P33 cdk2 ,P34 cdc2 ,cyclin B_1,cyclin E ,Mpm-2,Rb,PCNA

Summary: The inhibitory effects ofparthenolide (PTL) on angiogenesis induced by multiple myeloma (MM) cells in vitro, and the mechanism were investigated. Human MM line RPMI8226 cells were cultured in vitro. The effects of MM culture supernatant on the migration and tubule formation ability of human umbilical vein endothelial cells (HUVECs) treated with PTL were observed. By using Western blot, the expression of p65 and IκB-α in MM cells was detected. RT-PCR was used to assay the expression of VEGF, IL-6, MMP2 and MMP9 mRNA in MM cells. ELISA was used to measure the levels of VEGF and IL-6 in MM cell culture supernatant. The expression of MMP2 and MMP9 in MM cells was examined by immunohistochemistry. (1) In 3.5, 5.0, 7.5 and 10 μmol/L PTL groups the number of migrated cells was 310±56, 207±28, 127±21 and 49±10 respectively, which was significantly different from that in positive control group (598±47) (P<0.01). In 3.5 and 5.0 μmol/L PTL groups the areas of capillary-like structures were 0.092±0.003 and 0.063±0.002 mm2, significantly less than in positive control group (0.262±0.012 mm2) (P<0.01), but in 7.5 and 10 μmol/L PTL groups no capillary-like structures were found;(2) After treatment with different concentrations of PTL for 48 h, the expression of p65 protein was gradually decreased, while that of IκB-α was gradually enhanced with the increased concentration of PTL;(3) After treatment with 3.5,5.0, 7.5 and 10 μmol/L PTL for 48 h, the VEGF levels in the supematant were 2373.4±392.2,1982.3±293.3, 1247.0±338.4 and 936.5±168.5 pg/mL respectively, significantly different from those in positive control group (2729±440.0 pg/mL) (P<0.05). After treatment with 7.5 and 10 μmol/L PTL, the IL-6 levels in the culture supernatant were 59.6±2.8 and 41.4±9.8 pg/mL respectively, significantly lower than in positive control group (1287.3±43.5 pg/mL) (P<0.05);(4) RT-PCR revealed that PTL could significantly inhibit the expression of VEGF and IL-6 mRNA in MM cells, but not influence the expression of MMP2 and MMP9 mRNA.;(5) Immunohisto chemistry indicated that PTL had no significant effects on the expression of MMP2 and MMP9 protein in MM cells. It was concluded that the abilities of the culture supematant of MM cells treated with PTL to induce endothelial cells migration and tubule formation were significantly reduced, suggesting PTL could obviously inhibit the angiogenesis induced by MM cells. PTL could decrease NF-kappaB activity and significantly suppress the expression of VEGF and IL-6 mRNA and protein, which might contribute to the mechanism by which PTL inhibited the angiogenesis induced by MM cells.

Objective:To explore the technical focus of robotic-assisted laparoscopic surgery for the treatment of horseshoe kidney combined with renal tumor.Methods:The clinical data of a patient with horseshoe kidney combined with renal tumor treated by robot-assisted laparoscopic partial nephrectomy in the Second Hospital of Dalian Medical University in September 2021 were retrospectively analyzed. PubMed, CNKI, Wanfang and VIP databases were searched for all the literature on the use of robot-assisted laparoscopic nephrectomy or partial nephrectomy for the treatment of horseshoe kidney combined with renal tumor from the time of establishment to December 2022.Results:A total of 11 patients from 10 articles were retrieved and 12 patients were enrolled. Among the 12 patients, 4 cases used the retroperitoneal approach and 8 cases used the transperitoneal approach. Two cases were operated by traditional laparoscope, and the arteries were searched for and controlled before the robotic arm was placed to perform the partial nephrectomy and suture; and 10 cases were operated with the robotic-assisted laparoscopic approach throughout the whole procedure. Five cases of nephrectomy were performed on one side, and 7 cases were performed in the partial nephrectomy. Postoperative pathological diagnosis was clear cell carcinoma in 8 cases, chromophobe cell carcinoma in 1 case, eosinophilic cell carcinoma in 1 case, renal cell carcinoma in 1 case, and renal abscess in 1 case. The patient in the Second Hospital of Dalian Medical University was 38 years old female who was admitted to the hospital with a fever. After CT arteriography and three-dimensional reconstruction, robotic-assisted laparoscopic partial nephrectomy of right kidney and isthmus dissecting was performed. During the operation, tumor trophoblast vessels were ligated and dissected one by one by using single-use tissue closure clips, and the isthmus was dissected using endoscopic cutting anastomosis on the left side of the tumor, with the tumor edges sharply resected and completely dissected. The operation time was 240 min, without thermal ischemia time, and the bleeding volume was about 300 ml. The patient recovered well after the operation, and the postoperative pathological diagnosis was renal abscess.Conclusions:Robot-assisted laparoscopic treatment of horseshoe kidney combined with renal tumor is safe and effective, and has more advantages than traditional laparoscopic surgery. Preoperative CT arteriography or three-dimensional reconstruction examination should be applied to fully evaluate the variant vessels. The surgical access and plan should be decided according to the size and location of the tumor. The variant vessels should be properly handled during operation. The use of endoscopic cutting anastomosis to deal with the isthmus can be more conducive to the surgical operation.

Objective:To analyze the incidence and progression of overactive bladder(OAB)symp-toms following radical prostatectomy for prostate cancer patients and to identify related risk factors.Methods:A retrospective study was conducted on 263 local stage prostate cancer patients who underwent radical prostatectomy at Peking University Third Hospital from January 2013 to May 2017.Clinical base-line information,comprehensive imaging features,perioperative parameters,preoperative urinary control status,pathological diagnosis,and the incidence of OAB within one year postoperatively were collected and analyzed.In the imaging features,two parameters were defined:Bladder wall thickness(BWT)and bladder mucosal smoothness(BMS),which were used to predict the occurrence of OAB.Patients were evaluated based on their clinical baseline characteristics,including age,body mass index(BMI),co-morbidities,and prostate-specific antigen(PSA)levels.The imaging characteristics were assessed using preoperative MRI,focusing on BWT and BMS.Perioperative parameters included operative time,blood loss,and length of hospital stay.The OAB symptoms were assessed using the overactive bladder symptom score(OABSS)and the international prostate symptom score(IPSS).These scores were correlated with the postoperative incidence of OAB.Results:Among the 263 patients who underwent radical prostatecto-my,52(19.8％)exhibited OAB within one year postoperatively.Of the 40 patients with preoperative OAB symptoms,17(42.5％)showed remission postoperatively,while 23(57.5％)had persistent symptoms.Additionally,29 patients developed new-onset OAB,accounting for 55.77％of all postopera-tive OAB cases.Univariate analysis indicated that BWT,BMS,OABSS,and IPSS score were all associ-ated with the occurrence of postoperative OAB.Further multivariate analysis identified BMS as an inde-pendent risk factor for long-term OAB(P＜0.001).Conclusion:Long-term postoperative overactive bladder is a common complication following radical prostatectomy.The findings suggest that preoperative MRI measurements of bladder wall thickness and bladder mucosal smoothness during bladder filling phase can predict the risk of OAB occurrence postoperatively.Identifying these risk factors preoperatively can help in counseling patients about potential complications and in developing strategies to mitigate the risk of developing OAB after surgery.Early detection and management of these parameters might improve the quality of life for patients undergoing radical prostatectomy.

Objective:The transjugular or transfemoral approach is used as a common method for hepatic venous pressure gradient (HVPG) measurement in current practice. This study aims to confirm the safety and effectiveness of measuring HVPG via the forearm venous approach.Methods:Prospective recruitment was conducted for patients with cirrhosis who underwent HVPG measurement via the forearm venous approach at six hospitals in China and Japan from September 2020 to December 2020. Patients' clinical baseline information and HVPG measurement data were collected. The right median cubital vein or basilic vein approach for all enrolled patients was selected. The HVPG standard process was used to measure pressure. Research data were analyzed using SPSS 22.0 statistical software. Quantitative data were used to represent medians (interquartile ranges), while qualitative data were used to represent frequency and rates. The correlation between two sets of data was analyzed using Pearson correlation analysis.Results:A total of 43 cases were enrolled in this study. Of these, 41 (95.3%) successfully underwent HVPG measurement via the forearm venous approach. None of the patients had any serious complications. The median operation time for HVPG detection via forearm vein was 18.0 minutes (12.3~38.8 minutes). This study confirmed that HVPG was positively closely related to Child-Pugh score ( r = 0.47, P = 0.002), albumin-bilirubin score ( r = 0.37, P = 0.001), Lok index ( r = 0.36, P = 0.02), liver stiffness ( r = 0.58, P = 0.01), and spleen stiffness ( r = 0.77, P = 0.01), while negatively correlated with albumin ( r = -0.42, P = 0.006). Conclusion:The results of this multi-centre retrospective study suggest that HVPG measurement via the forearm venous approach is safe and feasible.