Functional constipation （FC） is a common functional disorder of the intestines， mainly characterized by reduced bowel movement frequency， difficulty in defecation， a sensation of incomplete evacuation， and hard stools， which severely affect patients’ quality of life. Research indicates that the pathogenesis of FC is closely related to gut microbiota dysbiosis and abnormal bile acid secretion. Bile acids， as endogenous natural laxatives， promote bowel movements by enhancing colonic secretion and regulating intestinal motility； meanwhile， gut microbiota influence colonic transit function by regulating the enteric nervous system， immune system， and their metabolic products. Based on an overview of the relationship between gut microbiota and bile acid metabolism， this article systematically reviews the current research status on the mechanisms of traditional Chinese medicine （TCM） in treating FC by regulating the balance of the gut microbiota-bile acid axis. It is found that single Chinese medicinal herbs （such as Atractylodes macrocephala）， isolated compounds （such as Platycodon grandiflorum polysaccharides）， herbal formulas （such as Shanger huang pill）， acupuncture， and moxibustion can up-regulate the abundance of beneficial bacteria， reshape the microbial structure， correct bile acid metabolism， and activate the Takeda G-protein receptor 5/farnesoid X receptor pathway to treat FC.

Objective To investigate the interaction between a novel antimicrobial compound,HL-J6,and penicillin-binding protein 1(PBP1)of Staphylococcus aureus.Methods With MRSA252 genomic DNA as the template and PBP1F and PBP1R as primers,the expression plasmid pET30a-pbp1-39-608 was constructed by amplifying the target gene fragment followed by cloning into the Nde I/Xho I restriction sites of the pET30a vector.Then the obtained plasmids were transformed into Escherichia coli for the expression of PBP1-39-608 protein,and the product was purified by affinity chromatography.The inhibitory effect of HL-J6 on the transpeptidase activity of PBP1-39-608 was measured using peptidoglycan side chain backbone peptide,with thiol ester analog S2d as the substrate.The affinity between HL-J6 and PBP1-39-608 was detected using microscale thermophoresis(MST),and the binding interaction was confirmed by cellular thermal shift assay(CETSA).Molecular docking and dynamics simulation were performed using AutoDock Vina and Desmond software,respectively,to elucidate the binding mode of HL-J6 with the PBP1-39-608 protein and the key amino acid residues involved.Results The recombinant plasmid pET30a-pbp1-39-608 was successfully constructed,and PBP1-39-608 protein was produced after induction and purified,yielding a protein with an approximate molecular mass of 65×103.HL-J6 inhibited the transpeptidase activity of PBP1-39-608 in a time-dependent manner(P<0.001).The dissociation constant Kd of the binding between HL-J6 and PBP1-39-608 was 64.92 μmol/L.Molecular docking results showed that HL-J6 bound to the active pocket of PBP1-39-608 by interacting with key residues such as ILE-348,ASN-370,THR-516 and PHE-423,with a binding score of-8.38 kcal/mol(<-5.00 kcal/mol).Dynamics simulation results indicated that the complex became stable after 50 ns.Conclusion HL-J6 effectively inhibits the transpeptidase activity of Staphylococcus aureus PBP1,and shows stable interaction with the protein.

Objective:To evaluate a self-designed intelligent robot-assisted minimally invasive reduction system in the reduction of unstable pelvic fractures by a cadaveric anatomic study.Methods:Ten unembalmed cadavers (7 male and 3 female ones) were used in this study. In each cadaveric specimen an unstable pelvic fracture was created in accordance with clinical case models (3 cases of type B1, 4 cases of type B2 and 3 cases of type C1 by the Tile classification). A self-designed intelligent robot-assisted minimally invasive reduction system was used to assist the reduction in the cadaveric models. Intraoperative registration and navigation time, autonomous reduction time, total operation time and reduction error were measured.Results:Effective reduction was completed in 10 bone models with the assistance of our self-designed intelligent robot-assisted minimally invasive reduction system. The time for intraoperative registration and navigation averaged 47.4 min (from 32 to 74 min), the autonomous reduction time 73.9 min (from 48 to 96 min), and the total operation time 121.3 min (from 83 to 170 min). The reduction error averaged 2.02 mm (from 1.67 to 2.62 mm), and the reduction results met the clinical requirements.Conclusion:Our self-designed intelligent robot-assisted minimally invasive reduction system is a new clinical solution for unstable pelvic fractures, showing advantages of agreement with clinical operative procedures, high reduction accuracy and operational feasibility, and reduced radiation exposure compared to a conventional operation.

Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.

Objective To investigate the change rules and its significance of erythrocytes surface molecule CD35 , CD58 and CD59 expression in recipients infected with cytomegalovirus (CMV)after renal transplantation. Methods Eighty-two recipients undergoing allogeneic renal transplantation were selected and divided into the negative (n=21 )and positive CMV groups (n=61 )based on the qualitative detection of CMV-pp65 antigen in peripheral blood. According to the results of CMV-pp65 (+)leucocyte count,all 61 patients in positive CMV group were further divided into low (n=55)and high active infection subgroups (n =6 ). Healthy adults were recruited into the normal control group (n =30 ). The expression levels of CMV-pp65 antigen,erythrocytes surface molecule CD35,CD58 and CD59 were measured by flow cytometry. Results Compared with normal control group,the expression levels of erythrocytes surface molecule CD35 , CD58 and CD59 in the positive CMV group were significantly down-regulated,and the CD35 and CD59 expression in the negative CMV group were considerably down-regulated (all P<0. 05 ). Compared with negative CMV group,the expression levels of CD58 and CD59 in the positive CMV group were significantly down-regulated (both P<0. 05 ). The expression levels of CD35 and CD59 in the high active infection subgroup were significantly lower than those in the low active infection subgroup (both P<0. 05 ). Conclusions The more severe active CMV infection after renal transplantation,the lower expression of erythrocytes surface molecule CD35,CD58 and CD59,hinting that red cell immune dysfunction is probably involved with active CMV infection.

Objective To observe the changes of serum homocysteine (Hcy)level of renal transplant recipients before and after renal transplantation,and assess the correlation between serum Hcy level and graft function.Methods Thirty-three recipients were included into the transplantation group,who underwent renal allograft transplantation in the Organ Transplant Institute of the 309 th Hospital of People's Liberation Army and had renal function recovered stably from January 2013 to June 2014.And 65 patients who were confirmed as chronic renal failure (CRF)by clinical examinations were included into the CRF group and 30 healthy people were included into the control group.A retrospective cross-sectional study was conducted on all of these subjects.Serum Hcy,serum creatinine (Scr)and blood urea nitrogen (BUN)levels of these three groups were compared.Serum Hcy and Scr levels of the transplantation group were continuously monitored before transplantation and at 3,7,14 and 21d after transplantation.The correlation between the changes of serum Hcy levels and the renal function before and after transplantation was assessed.Results Serum Hcy level of the CRF group was (25 ±10)μmol/L,which was significantly higher than (9 ±4)μmol/L of the control group and (15 ±9)μmol/L of the transplantation group in stable period,with statistical significance (all in P <0.001).Serum Hcy level of the transplantation group was significantly higher than that of the control group(P <0.001).Scr level of the CRF group,the transplantation group and the control group was(708 ±302)μmol/L, (98 ±23)μmol/L and (72 ±18)μmol/L,respectively.Scr level of the CRF group was significantly higher than those of the transplantation group and the control group (all in P <0.001).BUN level of the CRF group, the transplantation group and the control group was (18.1 ±5.9)mmol/L,(10.9 ±5.3)mmol/L and (4.9 ± 1.3) mmol/L, respectively. BUN level of the CRF group was significantly higher than that of the transplantation group and the control group (all in P <0.001),and BUN level of the transplantation group was significantly higher than that of the control group (P <0.001).With the improvement in renal function after transplantation,Scr and serum Hcy levels of the transplantation group deceased gradually.At 14 d after transplantation,Hcy level decreased to the minimum of (15 ±5)μmol/L.Compared with (25 ±10)μmol/L before transplantation,the difference had statistical significance (P <0.05).Within 14 d after transplantation, serum Hcy level of the transplantation group was positively correlated with Scr level (r =0.761,P <0.05). Conclusions Serum Hcy level of the renal transplant recipients is correlated with the graft function.The combined detection of serum Hcy and renal function index has certain guiding significance in the prevention of hyperhomocysteinemia and the early assessment of graft function.

Objective To study the effects of tacrolimus(Tac) concentrations on the number of NK cells and receptor expression in peripheral blood of renal transplantation receptors.Methods A total of 60 first-time kidney transplantation recipients in our institute from Dec.2007 to July 2009 were followed up.Tac maintenance immunosuppressive therapy was given to all recipients.The recipients were divided into low-concentration Tac group (6.84 + 1.72μg/L,n =30) and highconcentration Tac group ( 11.88 + 2.59 μg/L,n =30) according to concentrations of Tac.Twenty healthy volunteers served as controls.Before and 6 months after operation,concentrations of Tac were analyzed by using micro particle immunoassay chemiluminescent method.NK cells and their receptors (CD85j,CD158d,CD94 and NKG2D) were detected by using flow cytometry.The concentrations of soluble HLA-G5 were detected by using ELISA.Results The number of NK cells in lowconcentration Tac group and high-concentration of Tac group preoperatively was significantly reduced as compared with control group (P ＜ 0.05 ). The percentage and number of NK cells in low concentration Tac group and high-concentration Tac group at 6th month after operation were significantly reduced as compared with control group (P＜0.05).The number of NK cells in lowconcentration Tac group was significantly greater than in high-concentration Tac group (P＜ 0.05).There was no significant differende in the expression of CD85j,CD158,CD94 and NKG2D before operation between two groups(P＞0.05).The expression of CD85j and CD158d in two groups was increased,but that of CD94 and NKG2D was decreased at 6th month post-transplantation as comapred with that preoperation.In low-concentration Tac group,the expression of CD85j and CD158d was increased as compared with that in high-concentration Tac group (P＜0.05 ).Spearman correlation analysis revealed that the CD85j and CD158d expression had a positive correlation with sHLA-G5(P＜0.01 ),but the NKG2D had a negative correlation with sHLA-G5(P＜0.01 ).Conclusion There was correlation between the concentrations of Tac and NK cells count and NK receptors. Low concentrations of Tac can safely and effectively protect kidney function.The number of NK cells andtheir inhibitor receptors are increased in the recipients with low concentration of Tac.

The 26S proteasome is a proteolytic complex responsible for the degradation of the vast majority of eukaryotic proteins. Regulated proteolysis by the proteasome is thought to influence cell cycle progression, transcriptional control, and other critical cellular processes. A novel Schistosoma japonicum gene (GenBank Accession No. AY813725) proteasome alpha2 subunit (SjPSMA2) was cloned. Sequence analysis revealed that the ORF of SjPSMA2 gene contains 708 nucleotides encoding 235 amino acids, and the molecular weight was estimated to be 25.84 kDa. Real-time PCR analysis showed that this gene expressed in 7 d, 13 d, 18 d, 23 d, 32 d and 42 d schistosoma. The mRNA level of SjPSMA2 was lower in 7 d and 23 d schistosomulum than that in other stages. The SjPSMA2 cDNA fragment was subcloned into an expression vector pET28a(+) and transformed into E. coli BL21 (DE3) cells. After induction with IPTCQ the 30 kDa fusion protein was produced as included bodies. Western-blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation, and the protein in native could be detected. After immunization of BALB/c mice with the fusion protein, the reduction rates of worm counts and liver egg counts were 12.33% and 35.23%. ELISA results revealed that the vaccinated group showed a significant increase in the level of IgG antibody. This study provided an important basis for investigating the regulation mechanism of the proteasome during the development of Schistosoma japonicum.
Animals ; Antibodies, Helminth ; blood ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genes, Helminth ; Helminth Proteins ; genetics ; metabolism ; Immunization ; Liver ; parasitology ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Parasite Egg Count ; Proteasome Endopeptidase Complex ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Schistosoma japonicum ; genetics ; metabolism ; Vaccines, Synthetic ; immunology