Objective To investigate the immune regulatory effects of allogeneic bone marrowderived mesenchyrnal stem cells (MSCs) co-transplanted with islets. Methods The 18 diabetic mice were randomly divided into 3 groups: Diabetic group, without any transplantation; Islet transplantation group, in a sterile operation, with 10 μl purified islets (about 200 islets)transplantation to the left renal subcapsule of recipients; Islet + MSCs transplantation group, in addition to transplantation as the former group, 1 × 106 MSCs were given to the recipients via tail vein on 3, 2 and 0 days before islet transplantation. Blood glucose in recipient mice was monitored for 30consecutive days after transplantation. Pathological characteristics of left kidney were analyzed on the day 14 and 28 after transplantation. Th1/Th2, Tc1/Tc2, naive and memory T cells from peripheral blood and bone marrow-derived dendritic cells (DCs) were analyzed by multi-color flow cytometry.Results As compared with the islet transplantation group, blood glucose was significantly reduced,inflammatory cell infiltration decreased in the place of transplantation, graft survival prolonged, the number of Th1 and Tc1 cells was obviously reduced, the number of Th2 and Tc2 cells increased, the ratio of Th1/Th2 and Tc1/Tc2 cells was significantly decreased, the naive and memory T cells were significantly inhibited, the maturity of DCs and the secretion of interleukin-12 decreased in the islet transplantation group. Conclusion Through the immunomodulation on T-cell and DCs function,MSCs can alleviate graft verse host disease and prolong allograft survival.

AIM:To investigate the role of B cells in CD45RB antibody-induced transplantation immune toler-ance.METHODS:Single cell suspension was made from the spleen of BALB/c nude mice disposed by CD45RB antibod-y, then mixed cultured with T cells of BALB/c mice and spleen cells of C57BL/6 mice.The Th1, Th2, Treg and Tm cells were monitored by flow cytometry during the culture process .The skin graft model was set up with B 6.μMT-/-mice as re-ceptors and BALB/c mice as donors.CD45RB antibody was intraperitoneally injected into the receptors after transplantation and then CD3+CD45RBhi cells were detected by flow cytometry .In another mixed lymphocyte culture , CD45RB antibody was added, and then B cells were isolated and injected into B6.μMT-/-mice through the tail vein.The heart transplanta-tion model was established with B 6.μMT-/-mice as receptors and BALB/c mice as donors, and then the survival and the migration of B cells to the thymus were observed .RESULTS:When T lymphocytes were co-cultured with B lymphocytes treated with anti-CD45RB monoclonal antibody (mAb) in vivo, the percentages of Th2 and Treg cells were up-regulated and Th1 cells were down-regulated, but Tm cells were not altered as compared with the control .In vivo without B lympho-cytes, anti-CD45RB mAb also down-regulated the expression of CD45RB in T lymphocytes.The reduction was faster and the percentage of CD3 +CD45RBhi T cells was not altered as compared with the control .The B lymphocytes treated with an-ti-CD45RB mAb in vitro prolonged the lifetime of receptor in heart transplantation model but failed to induce complete toler -ance.After recieving B cells treated with anti-CD45RB mAb and allogeneic heart transplantation , B cells migrated to the thymus in B6.μMT-/-mice.CONCLUSION:B lymphocytes play a definite role in the transplantation immune tolerance induced by anti-CD45RB mAb through their affection on T-cell subgroups and also in the central tolerance .However, the induction of immune tolerance can not only rely on B cells .

Due to the extensive use of xylooligosaccharides (XOS) as functional food ingredients,many inferior goods and even adulterants are generally found in the market,which may pose a health hazard to certain populations.Chromatography method such as high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) is traditionally applied for the quality analysis of XOS.However,it is time consuming due to the prolonged separation and pre-or post-derivatization procedure.In this study,a fast saccharide mapping method based on matrix-assisted laser desorption/time-of-flight mass spectrometry (MALDI-TOF-MS) was developed for the quality consistency analysis of 22 batches of XOS collected from different manufacturers in China.The time needed for saccharides analysis using MALDI-MS was less than 30 min for one plate,at least 6 times faster than that by the traditional HPTLC chromatography method.In addition,MALDI-MS possessed higher resolution for XOS with DP4-DP7 based on the difference of m/z,which is hardly separated using HPTLC.The results showed that XOS were present only in samples XY01-XY11,samples XY12-XY14 only consisted of hex oligo-saccharides,and samples XY15-XY22 were free of oligosaccharides.These indicate that the quality consistency of XOS products in the China market was poor,which should be carefully investigated.

Objective To explore the effects and action mechanism of miR-181c-5p on biological behaviors of pros-tate cancer cells.Methods The pathological relationship between BIRC5,miR-181c-5p and prostate cancer was analyzed based on prostate cancer data in TCGA database.The target binding site of miR-181c-5p and BIRC5 was analyzed by miRNA target gene prediction database,and was verified by double luciferase activity assay.The ex-pression of BIRC5 protein in miR-181 c-5p overexpression cells was detected by Western blot.The prostate cancer cells PC3 and DU145 were selected to construct cell line with miR-181c-5p overexpression(miR-181c-5p group)and its negative control(miR-NC group),and qRT-PCR verification was conducted.The cells proliferation[opti-cal density at 450 nm site(OD450 nm)]was detected by CCK-8.Distribution of cell cycles and apoptosis rate were detected by flow cytometry.Expressions of proliferation and apoptosis related proteins were detected by Western blot.The cell line with miR-181c-5p/BIRC5 overexpression was constructed(miR-181c-5p+BIRC5 group).Cells growth,distribution of cell cycles,apoptosis rate and expressions of related proteins were detected by the a-bove methods.Results The expression of BIRC5 was up-regulated in prostate cancer tissues,and it was higher in patients with high tumor invasion,lymph node metastasis and recurrence.Patients exhibiting high expression of BIRC5 demonstrated poor survival rates.The expression of miR-181c-5p was down-regulated in prostate cancer tis-sues.The level of miR-181c-5p was negatively correlated with BIRC5 level,and miR-181c-5p could inhibit BIRC5 expression.In PC3 and DU145,miR-181c-5p level in miR-181c-5p group was higher than that in miR-NC group(P＜0.05);OD450 nm and percentage of S-phase cells were lower than those in miR-NC group(P＜0.05),per-centage of cells in G0/G1 phase;apoptosis rate and expressions of BAX,caspase-3 and PARP proteins were higher than those in miR-NC group(P＜0.05);expressions of CDK2,CCNB1 and BCL-2 proteins were lower than those in miR-NC group(P＜0.05).The expression of BIRC5 protein and OD450 nm in miR-181 c-5p+BIRC5 group were higher than those in miR-181c-5p group(P＜0.05),percentage of cells in G0/G1 phase was lower than that in miR-181c-5p group(P＜0.05);percentage of S-phase cells was higher than that in miR-181c-5p group(P＜0.05);apoptosis rate was lower than that in miR-181c-5p group(P＜0.05);expressions of CDK2,CCNB1 and BCL-2 proteins were higher than those in miR-181c-5p group;expressions of BAX,caspase-3 and PARP proteins were lower than those in miR-181 c-5p group.Conclusion miR-181-5p can inhibit the proliferation of human pros-tate cancer cells by targeting BIRC5,block cells in G0/G,phase and promote cells apoptosis.

Objective To explore the clinical efficacy of high-frequency electric snare device under elec-tronic laryngoscope in the treatment of epiglottic cyst.Methods A total of 100 patients with definitely diag-nosed epiglottic cyst receiving outpatient operation treatment in the First Affiliated Hospital of Gannan Medi-cal University from April 2021 to March 2023 were selected as the research subjects and included into the ob-servation group and control group according to the visiting order,50 cases in each group.The observation group was treated with high-frequency electric snare for epiglottic cyst resection under electronic laryngo-scope,and the control group was treated with laryngeal tissue forceps under electronic laryngoscope for uncov-ering operation of epiglottic cyst.The operation time,intraoperative blood loss volume,pain degree within postoperative 24 h,pain duration,complete resection rate and recurrence rate in postoperative 3 months were compared between the two operation methods.Results The operation time,intraoperative blood loss volume,VAS score within postoperative 24 h and pain duration in the observation group were significantly less than those in the control group,the complete resection rate was significantly higher than that in the control group,and the differences were statistically significant (P<0.05).The two groups were followed up for 3 months. Only 2 cases in the control group relapsed,which were epiglottic multiple cyst,and the recurrence rate had no statistical difference between the two groups (P>0.05).No dyspnea,massive bleeding and epiglottic adhesion appeared.Conclusion The high-frequency electric snare device under electronic laryngoscope for treating epi-glottic cyst has the advantages of short operation time,less blood loss,postoperative light pain and high com-plete resection rate.

OBJECTIVETo investigate whether transplantation of autologous peripheral blood CD34+ stem cells is a viable approach for treating patients with advanced cirrhosis,which is currently hindered by a shortage in liver donors.

METHODSA total of 100 patients with advanced cirrhosis and who had failed to respond to conservative therapy were recruited for transplantation of autologous peripheral blood CD34+ stem cells.The success of transplantation was investigated 6-and 12-months later by measuring markers of liver biosynthesis function (coagulation,albumin level,indocyanine green clearance,Child-Pugh score) and assessing pathological changes (Knodell score) and morphologic changes in the liver tissue.Complications were also recorded during follow-up.

RESULTSThe 1-year cumulative survival rate was 100%. Fifty-two patients with massive ascites showed gradual reduction and disappearance of the ascites.Four patients experienced upper gastrointestinal bleeding and three patients developed with hepatic encephalopathy (I-II degree) at 3 months post-transplantation.All patients showed significantly improved liver biosynthesis function,liver elasticity and Knodell score after transplantation (P less than 0.05).

CONCLUSIONAutologous peripheral blood CD34+ stem cell transplantation is a safe and effective treatment for advanced cirrhosis,and has high cost-benefit since it improves liver function,liver histology,and quality of life.

Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features.Therefore,quantitatively quality control of polysaccharides based on their chemical charac-teristics is important for their application in biomedical and functional food sciences.However,poly-saccharides are mixed macromolecular compounds that are difficult to isolate and lack standards,making them challenging to quantify directly.In this study,we proposed an improved saccharide mapping method based on the release of specific oligosaccharides for the assessment of Hericium eri-naceus polysaccharides from laboratory cultured and different regions of China.Briefly,a polysaccharide from H.erinaceus was digested by β-(1-3)-glucanase,and the released specific oligosaccharides were labeled with 8-aminopyrene-1,3,6-trisulfonic-acid(APTS)and separated by using micellar electrokinetic chromatography(MEKC)coupled with laser induced fluorescence(LIF),and quantitatively estimated.MEKC presented higher resolution compared to polysaccharide analysis using carbohydrate gel elec-trophoresis(PACE),and provided great peak capacity between oligosaccharides with polymerization degree of 2(DP2)and polymerization degree of 6(DP6)in a dextran ladder separation.The results of high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector(HPSEC-MALLS-RI)showed that 12 h was sufficient for complete digestion of polysaccharides from H.erinaceus.Laminaritriose(DP3)was used as an internal standard for quantifi-cation of all the oligosaccharides.The calibration curve for DP3 showed a good linear regression(R2＞0.9988).The limit of detection(LOD)and limit of quantification(LOQ)values were 0.05 μg/mL and 0.2 μg/mL,respectively.The recovery for DP3 was 87.32(±0.03)％in the three independent injections.To sum up,this proposed method is helpful for improving the quality control of polysaccharides from H.erinaceus as well as other materials.

Hepatic alveolar echinococcosis (HAE) is a common zoonotic endemic parasitic disease in western China. It lacks of typical clinical manifestations in the early stage, and symptoms become prominent during the end stage, with an alarmingly high mortality rate. Among the treatment of end-stage HAE (es-HAE), orthotopic liver transplantation is almost the only radical treatment due to insufficient remnant liver volume, uncontrollable bleeding and difficulty in vascular reconstruction in vivo. However, the shortage of donor liver and long-term postoperative use of immunosuppressants limit its application. The introduction of ex vivo liver resection and autotransplantation (ELRA) resolves this dilemma and significantly broadens the indications of es-HAE. In addition, multiple centers in China have optimized and modified ELRA to further improve the treatment system of es-HAE. At present, liver transplantation (including ELRA) of es-HAE remains a hot topic for clinicians. In this article, orthotopic liver transplantation, ELRA, auxiliary ELRA and other surgical treatment of es-HAE were reviewed, aiming to further enhance the diagnosis and treatment of es-HAE and improve clinical prognosis of the patients.

Anti-seizure medications (ASMs) are the main therapy for epilepsy.There are many kinds of ASMs with complex mechanism of action, so it is difficult for pharmacists to examine prescriptions.This paper put forward some suggestions on the indications, dosage forms/routes of administration, appropriateness of usage and dosage, combined medication and drug interaction, long-term prescription review, individual differences in pathophysiology of children, and drug selection when complicated with common epilepsy, for the reference of doctors and pharmacists.

Objective: To further observe the efficacy of combined transplantation of islet and bone marrow mesenchymal stem cells (BMSC) in diabetic rats, PET-CT was used to trace cells in vivo to determine the homing and distribution of cells in vivo. Methods: Streptozotocin (STZ)was used to construct a rat model of diabetes mellitus. BMSC could be isolated and cultured by full adherence method; islets were isolated by collagenase; Islets and BMSC were labeled with 18F-FDG in vitro. Diabetic rats were randomly divided into 4 groups, 15 rats in each group: A, Control group; B, Stem cell transplantation group; C, Islet Transplantation group; D, Combined transplantation group, a total of four groups, all transplanted through portal vein, PET-CT tracing the distribution of cells transplanted into the body.7 days after transplantation, the livers of each group were taken, and the homing and distribution of transplanted cells were detected by immunofluorescent staining.The SUV was calculated by the analysis of variance of random block, and the difference between groups was compared by t-test. Results: PET-CT results showed that BMSC were mainly distributed uniformly in the right liver, and the islets of the pancreas were mainly clustered in terminal branches of hepatic portal vein, and BMSC were around the islets of pancreas, but there was no obvious development in the liver of the control group. Conclusions PET-CT can directly reveal the distribution of islets and BMSC in liver after transplantation through portal vein.