Objective To determine the relationship between blood biochemical indexes and severity of NAFLD by establishing various models of NAFLD with different degrees in rabbits. Methods Thirty-two healthy rabbits were randomly divided into normal control (n=8) and three experimental groups(n=24). Fatty liver animal models were established by giving high fat,high sugar diet. The serum TG,TC,AST,ALT and pathological evaluation were detected respectively after 4,6 and 8 weeks. Results The rabbits developed hyperlipidemia and slight fatty liver after four weeks. Along with the worsening of fat liver,the serum TG level kept rising.Conclusion The blood biochemical indexes can serve as a reliable technique for diagnosing fatty liver disease in clinical.

AIM To study the growth inhibition of adenosine triphosphate (ATP) and adenosine (ADO) on cultured tumor cells in vitro. METHODS MTT assays were used to determine the inhibition of proliferation of ATP and ADO on several tumor cell lines, including human squamous esophageal carcinoma cells TE-13, human stomach carcinoma cells HGC-27 and human erythroleukemia cells K562. The morphological changes of ATP and ADO on the cell lines were observed under light microscope. RESULTS ATP and ADO produced a certain inhibition effect on TE-13, HGC-27, K562 cells at different concentration between 0.01～1.0 mmol?L -1. The maxium inhibition fraction of TE-13, HGC-27, K562 cells exposed to ATP for 72 h was 80.52%, 58.67% and 45.07%, respectively; and to ADO for 72 h, was 74.03%, 52% and 30.99%, respectively. Under light microscope, the tumor cells exposed to higher concentration(1 mmol?L -1) of ATP and ADO displayed morphological changes of apoptosis. CONCLUSION These results suggested that ATP has a certain cytotoxic effect on several tumor cell lines, it might partly be related to ADO. Its mechanism might involve apoptosis.

Extracellular adenosine triphosphate (ATP) and adenosine are important signaling molecules in both intracellular and extracellular microenviroments of cells. They exert cytoxic and apoptotic effects in some tissues such as tumor. The proapoptotic mechanisms of adenosine and its receptors are involved in Adenylyl cyclase(AC), cAMP, Ca 2+ , etc. Apoptosis induced by ATP is as sociated with several purinergic receptors including P2X_1, P2X_2, P2X_7, P2Y_1, P2Y_2, et al, it is also involved in FasL/FasR, T cell receptor(TCR), and gene regulations of bcl-2 and bax.

Liver transplantation is a standard life-saving procedure for end-stage liver diseases.The therapeutic potential of this procedure may be limited by post-operative infectious complications.A better understanding on the common important infectious complications may improve the life quality and survival rate after liver transplantation.In this article,we review the progress on infectious complications after liver transplantation,with particular emphasis on risk factors,clinical manifestations,diagnostic methods,prevention measures and specific treatments for bacterial,fungal,cytomegalovirus infections.

Objective To observe any effect of low intensity pulsed ultrasound (LIPUS) on the expression of integrin-focal adhesion kinase (FAK)-mitogen-activated protein kinase (MAPK) mechanochemical transduction pathway-related proteins in the chondrocytes of rabbits with knee osteoarthritis (OA).Methods Of the 18 New Zealand white rabbits selected for the study,twelve received knee anterior cruciate ligament transection to model OA.The remaining 6 rabbits served as normal controls.At the 4th week after modeling the rabbits were sacrificed and chondrocytes were isolated and cultured in vitro.All the cultured cells were randomly divided into three groups:a normal control group (NC),an OA model group (OA) and an OA model plus LIPUS group (OA + LIPUS).When the cells had been cultured to the 2nd passage,the NC group and OA group cells had received no treatment.The OA + LIPUS group cells were exposed to 40 mW/cm2 of LIPUS for 20 min,once a day for 6 days.The expression of collagen protein type Ⅱ,aggrecan,MMP-13,integrin β1 p-FAK and p-p38,p-ERK1/2 and p-JNK Mapks were detected by Western blotting.Results Compared with the NC group,the expression of collagen type Ⅱ and aggrecan was significantly lower in the OA and OA + LIPUS groups,with more significantly lower expression of collagen type Ⅱ and aggrecan in the OA group than in the OA + LIPUS group.Compared with the NC group,the expression of MMP-13 was significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA group.Compared with the NC group,the expression of integrin β1 and p-FAK was also significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA + LIPUS group.Compared with the NC group,the expression of p-p38,p-ERK1/2 and p-JNK was also significantly higher in the OA group,but compared with the OA group,the expression of those kinases was,on average,significantly lower in the OA + LIPUS group.Conclusions LIPUS can inhibit the degradation of collagen type Ⅱ and aggrecan,and inhibit the expression of MMP-13,p-p38,p-ERK1/2 and p-JNK in OA chondrocytes,at least in vitro.At the same time,LIPUS can increase the expression of integrin β1 and p-FAK.The results show that LIPUS may activate an integrin-FAK-MAPK mechanochemical transduction pathway to induce changes in the extracellular matrix of chondrocytes.

Objective To observe the effect of low intensity pulsed ultrasound (LIPUS) on chondrocytes co-cultured with infrapatellar fat pads.Methods Twenty-four infrapatellar fat pads and cartilages from female knee trauma patients aged between 25 and 35 were cut and stained using hematoxylin-eosin staining.Chondrocytes were isolated from part of the integrated surface of the cartilages to be cultured in vitro.They were then randomly divided into a normal chondrocyte group (the control group),a normal chondrocyte+FCM (fat conditioned medium) group (the model group),a normal chondrocyte+ FCM + LIP US group (the treatment group),and a normal chondrocyte+ FCM + LIPUS + GRGDSP (an integrin inhibitor) group (the inhibited group).The treatment group and inhibited group received LIPUS at 40 mW/cm2 for 20 min once a day,while the other groups received sham LIPUS treatment.Five days later,the cells were collected and western blotting was used to examine the expression of type Ⅱ collagen (COL2),aggrecan (Acan),matrixmetalloproteinase (MMP)-13,integrin β1,phosphorylation-focal adhesion kinase (p-FAK) and phosphorylation p38 (p-p38).Results Western blotting showed that compared with the control group,the expression of COL2 and Acan was significantly lower in the model group,but that of MMP-13,integrin β1,p-FAK and p-p38 was significantly higher.As compared with the model group,the expression of COL2,Acan,integrin β1 and p-FAK was significantly higher,and that of MMP-13 and p-p38 was significantly lower in the treatment group.The expression of COL2,Acan,MMP-13,integrin β1,p-FAK and p-p38 showed no significant difference between the inhibited and model groups,but that of COL2,Acan,MMP-13,integrin β1,p-FAK and p-p38 was significantly different between the control and treatment groups.Conclusions LIPUS provides a protective effect on chondrocytes through inhibiting the expression of MMP-13 induced by adipokines and the degradation of COL2 and Acan through activating the integrin-FAK-p38 MAPK pathway.

Objective To investigate the role and mechanism of ABI family member 3-binding pro-tein(ABI3BP)in dysfunction of endothelial progenitor cells(EPC)induced by angiotensin Ⅱ(Ang Ⅱ).Methods EPC were divided into sh-RNA negative control(sh-NC)group(transfected with LV-scramble-shRNA+PBS),sh-ABI3BP group(transfected with LV-ABI3BP-shRNA+PBS),sh-NC+Ang Ⅱ group(transfected with LV-scramble-shRNA+Ang Ⅱ)and sh-ABI3BP+Ang Ⅱ group(transfected with LV-ABI3BP-shRNA+Ang Ⅱ)to investigate the effect of silencing ABI3BP on the dysfunction of EPC induced by Ang Ⅱ.Transwell assay,adhesion assay,Matrigel tube formation assay,and TUNEL staining were performed respectively to detect the migration,adhesion and tube formation abilities and cell apoptosis in above cell groups.The expression chan-ges in integrin-β1/FAK/P53 signaling pathway were detected by Western blotting.Results Com-pared to the sh-NC group,the sh-NC+AngⅡ group showed significant decreases in the numbers of migratory cells,adhesion cells,and tubule formation,along with increases in the apoptotic rate and the expression levels of Integrin β1,p-FAK,and P53(P＜0.05).While,the sh-ABI3BP+AngⅡ group had obviously more migratory cells(88.67±8.33 vs 62.33±7.37 units,P＜0.05),adhe-sion cells(104.33±6.03 vs 68.33±10.05 units,P＜0.05),and tubule formation(36.33±3.21 vs 19.33±3.06 units,P＜0.05),while decreased apoptotic rate and expression levels of integrin-β1,p-FAK and P53 protein when compared with the sh-NC+AngⅡ group(P＜0.05).Conclusion Ang Ⅱ can up-regulate the expression of ABI3BP,and knockdown of ABI3BP can improve Ang Ⅱ-induced EPC dysfunction,which may be related to its inhibition on integrin-β1/FAK/P53 signa-ling pathway.

Objective To investigate any protective effect of low-intensity pulsed ultrasound (LIPUS) and pioglitazone on chondrocytes in osteoarthritic patients using the pathway from peroxisome proliferator-activated γreceptor (PPARγ) to nuclear factor kappa B (NF-κB) to inducible nitric oxide synthase (iNOS).Methods Normal chondrocytes of 24 healthy adult New Zealand white rabbits were extracted and divided into a normal group,a lipopolysaccharide (LPS) group,a LIPUS group (LPS+LIPUS) and a pioglitazone group (LPS+pioglitazone),each of 6 using a random number table.Each group was given the intervention their names implies.The levels of tumor necrosis factor-α (TNF-α),leptin (LEP) and nitric oxide (NO) in the chondrocytes were detected using enzyme-linked immune sorbent assays.The expression of type Ⅱ collagen (COL2) in the chondrocytes of each groups was detected using immunocytochemistry and fluorescent staining.The mRNA and protein expressions of PPARγ,NF-κB and iNOS were detected using reverse transcription polymerase chain reactions and western blotting respectively.Results Compared with the LPS group,the average level of TNF-α,LEP and NO in the LIPUS and pioglitazone groups was significantly lower,with the levels in the pioglitazone group significantly lower than in the LIPUS group.Compared with the LPS group,COL2 expression in the LIPUS group was significantly greater.The mRNA and protein expressions of PPARγ in the chondrocytes in the LIPUS and pioglitazone groups were significantly higher than those in the LPS group.Compared with the LPS group,the mRNA and protein expressions of NF-κB and iNOS in the pioglitazone and LIPUS groups were significantly lower,with the pioglitazone group's levels significantly below those of the LIPUS group.Conclusion LIPUS and pioglitazone may promote anti-inflammatory action and COL2 synthesis in chondrocytes through the PPARγ/ NF-κB/iNOS pathway and play a protective role,at least in rabbits.

Objective To investigate the effects of icariin on high glucose-induced autophagy and apoptosis of podocytes,and the regulating effects on mammalian target of rapamycin(mTOR)/serine-threonine kinase(Akt)/cyclic adenosine monophosphate response element binding protein(CREB)pathway.Methods The mouse podocytes MPC5 were taken and divided into five groups:normal control group(5.5 mmol·L-1 glucose),high glucose group(30 mmol·L-1 glucose),icariin group(30 mmol·L-1glucose+5 μmol·L-1icariin),GDC-0349 group(30 mmol·L-1glucose+50 μmol·L-1 GDC-0349),icariin+GDC-0349 group(30 mmol·L-1 glucose+5 μmol·L-1 icariin+50 μmol·L-1 GDC-0349).Cultured for 48 hours,the tetramethylazozolium salt method was used to detect the viability of MPC5 cells;acridine orange staining was used to observe the autophagy of MPC5 cells;apoptosis of MPC5 cells was detected by flow cytometry;Western blotting was used to detect the expression of autophagy[microtubule associated protein one light chain 3(LC3)II,LC3Ⅰ,autophagy-related protein(Beclin-1)],apoptosis[Bcl-2 related X protein(Bax),B cell lymphoma-2(Bcl-2)]and mTOR/Akt/CREB pathway-related proteins of MPC5 cells.Results Compared with the normal control group,the cell viability,expression levels of Bcl-2,phosphorylated mTOR(p-mTOR)/mTOR,phosphorylated Akt(p-Akt)/Akt,phosphorylated CREB(p-CREB)/CREB protein of MPC5 cells in the high glucose group were significantly decreased(P＜0.05),the autophagy ability was enhanced,the autophagosome showed orange fluorescence,and the apoptosis rate,LC3Ⅱ/LC3Ⅰ,Beclin-1,Bax protein expression levels were significantly increased(P＜0.05).Compared with the high glucose group,the cell viability,LC3Ⅱ/LC3Ⅰ,Beclin-1,Bcl-2,p-mTOR/mTOR,p-Akt/Akt,p-CREB/CREB protein expression levels of MPC5 cells in icariin group were significantly increased,the autophagy ability was further enhanced,the number of autophagosomes was increased,the autophagosomes showed brick red fluorescence(P＜0.05),the apoptosis rate and Bax protein expression level were significantly decreased(P＜0.05),and the cell viability,LC3Ⅱ/LC3Ⅰ,Beclin-1,Bcl-2,p-mTOR/mTOR,p-Akt/Akt and p-CREB/CREB proteins expression levels of MPC5 cells in GDC-0349 group were significantly decreased,the autophagy ability was weakened,the number of autophagosomes was reduced,the autophagosomes showed orange fluorescence(P＜0.05),and the apoptosis rate and Bax protein expression level were significantly increased(P＜0.05);icariin+GDC-0349 could reverse the effect of icariin on high glucose induced MPC5 cells(P＜0.05).Conclusion Icariin promotes elevated glucose-induced podocyte autophagy and inhibits apoptosis by activating the mTOR/Akt/CREB pathway.

Objective:To compare the efficacy and safety of standard treatment with or without adjuvant chemotherapy in patients with highly malignant non-metastatic prostate cancer.Methods:In this prospective non-randomized controlled study, consecutive non-metastatic prostate cancer patients with pathologically proven Gleason score of 9-10 or Gleason score of 5 admitted to Peking University First Hospital were enrolled. Four to six cycles of chemotherapy using docetaxel ± carboplatin regimen were added or not after standard radical therapy. The primary end point was 5-year event-free survival (EFS), and the secondary end points were distant metastasis-free survival (MFS), overall survival (OS), and treatment-related adverse events. The survival curve was drawn by Kaplan-Meier method. The differences between two groups were analyzed by log-rank test.Results:A total of 176 patients were consecutively enrolled from November 2019 to January 2022 of which 138 patients received only standard radical therapy (control group), and 38 patients received adjuvant chemotherapy after standard radical therapy (chemotherapy group). The median follow-up time was 13.4 (2.0-34.0) months. All patients survived. The 30-month EFS rates in the chemotherapy and control groups were 100% and 85.6%, respectively ( P=0.064). There were no events in the chemotherapy group, while there were 12 cases of events in the control group, including 6 cases of biochemical recurrence and 6 cases of imaging progression. The 30-month MFS rates in two groups were 100% and 91.9%, respectively ( P=0.205). After the 1 vs. 2 propensity score matching, the EFS and MFS rates in two groups were 100% vs. 85.7% ( P=0.056), and 100% vs. 92.2% ( P=0.209), respectively. The incidence rates of grade 2 and above urinary toxicity in the chemotherapy and control groups were 2.6% and 7.2% ( P=0.354), respectively. The incidence rates of grade 2 and above rectal toxicity were 5.3% and 5.1% ( P=0.711), respectively. Grade 3 and above chemotherapy-related toxicity in the chemotherapy group were leukopenia (31.6%), thrombocytopenia (2.6%) and alopecia (13.2%). Conclusion:The addition of adjuvant chemotherapy after standard radical therapy tends to improve the overall EFS of patients with highly malignant prostate cancer, and the adverse effects are tolerable, which should be confirmed by long-term follow-up results.