AIM:To investigate whether mitochondrial membrane potential (ΔΨm ) and the mitochondrial ap-optotic pathway are involved in the protective mechanism of Panax quinquefolium saponin ( PQS) against cardiomyocyte ap-optosis after ischemia/reperfusion ( I/R) injury in rat myocardium .METHODS: Ninety healthy male SD rats were ran-domly divided into sham group , I/R group, PQS (200 mg· kg -1 · d-1 ) +I/R group, cyclosporine A ( CsA) group, CsA (10 mg· kg-1 ) +I/R group and PQS +CsA +I/R group.The model of myocardial I/R injury in vivo was established by ligating the left anterior descending artery ( LAD) for 30 min followed by 120 min of reperfusion in the rats .The serum ac-tivity of lactate dehydrogenase ( LDH ) was measured by automatic chemistry analyzer .The myocardial infarct size was measured by Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC) staining.Cardiomyocyte apoptosis was detected by in situ TDT-mediated dUTP nick end labeling (TUNEL).The protein levels of Bcl-2, Bax, cleaved caspase-3 and cytosol-ic cytochrome C were determined by Western blotting .ΔΨm was measured by laser scanning confocal microscopy and fluo-rescence microplate reader .RESULTS:Compared with I/R group, the serum content of LDH ,the infarction size in PQS+I/R group, CsA+I/R group and PQS +CsA+I/R group and the myocardial apoptotic index were decreased .Compared with I/R group, the fluorescence intensity of mitochondria after JC-1 staining was enhanced in PQS +I/R group, CsA+I/R group and PQS+CsA+I/R group, and the relative fluorescence units (RFU) ofΔΨm were improved in those 3 groups. In PQS+I/R group, CsA+I/R group and PQS+CsA+I/R group, the protein expression of Bcl-2 was increased, and that of Bax was decreased compared with I/R group.Moreover, in those 3 groups, the protein levels of cleaved-caspase-3 and cytosolic cytochrome C were decreased compared to I /R group, respectively.CONCLUSION:PQS attenuates myocardial injury and cardiomyocyte apoptosis during I /R, and the protective mechanisms of PQS were associated with the modulation ofΔΨm and the inhibition of mitochondrial apoptosis pathway .

Objective To compare the effects of 2 vascular carriers, arteriovenous loop and arteri-ovenous bundle, on inducing angiogenesis in coral scaffold of vascularized tissue-engineered bone in animal models.Methods Thirty-six adult male New Zealand rabbits were randomized into 2 even groups.In group A, an arteriovenous loop (AVL) was formed by microsurgical anastomosis at the proximal ends between the femoral poptiteal artery and vein, and placed in the circular side groove of the coral block (6 mm × 8 mm × 10 mm) .In group B, flow-through vessels bundles of both femoral artery and vein were placed in the side grooves of the coral block.All the implants in 2 groups were wrapped by a micro-porous expand-ed-polytetrafluoroethylene (ePTFE) membrane, and fixed subcutaneously by suturing.Evaluation methods included gross morphological observations, histological examinations, India ink perfusion and vascular casting after 2, 4, 6 weeks.The density of blood vessels was analyzed by the statistical software SPSS 10.0.Results All the corals were encased by newly formed fibrovascular tissues in 2 groups.Ink-stained vessels distributed the surfaces and side grooves, and invaded the interspaces of corals.The degree of vascularization increased over the course of experiment.Blood vessel density demonstrated a significant continuous increase between 2 and 6 weeks after implantation in group A.The mean value of blood vessel density in group A (2 weeks 276.60±4.67, 4 weeks 517.20±10.66, 6 weeks 707.00 ±11.87) was significantly higher than in group B (2 weeks 153.60 ±7.16, 4 weeks 269.40±6.80, 6 weeks 279.20±6.53) (P <0.01).Vascular casting showed that in group A, significant blood vessels sprouted from all areas of the loop, espe-cially at the entrance of the arteriovenous pediele where the small tubes were densely interconnected.In group B, however, no blood vessels sprouted from the arteriovenous bundles and only some small vessels grew from the entrance and exit.Conclusions A vascularized coral model can be constructed by inserting an ar-teriovenous loop or an arteriovenous bundle, useful in vascular bone tissue engineering.The former, however, have stronger abilities to induce angiogenesis than the latter.

Objective To investigate the mechanisms of hypoxia inducible factor-2 alpha (HIF-2a) regulating human umbilical vein endothelial cells (HUVECs) under hypoxic conditions.Methods The experimental study was adopted.(1) HUVECs in logarithmic growth phase were taken:HUVECs without any disposals as control group,HUVECs with shRNA transfection control as shRNA control group,HUVECs with HIF-2α shRNA transfection as HIF-2α shRNA group and HUVECs with HIF-2α shRNA transfection then added rhAng-2 as HIF-2α ± rh-Ang-2 group.(2) Western blot testing:the expressions of Ang-2 and HIF-2α proteins in HUVECs were cultured under hypoxia conditions at 0,2,4,8,12,16,20 hours,and the levels of which were detected in the control group,shRNA control group and HIF-2α shRNA group.(3) Enzyme-linked immunosorbent assay(ELISA):the level of Ang-2 protein in supernatant of HUVECs was detected in the control group,shRNA control group and HIF-2α shRNA group.(4)The amounts of endothelial cell tubes in HUVECs among the 4 groups were detected by tube formation experimental testing.(5) Transwell method was performed to detect the amounts of cells migration in HUVECs and hepatoma cells SMMC-7721 migration intervened by supernatant of HUVECs among the 4 groups.Measurement data with normal distribution were presented as x ± s,repeated measurement data were analyzed by the repeated measures ANOVA,comparison among groups and pairwise comparison were conducted respectively by the one-way ANOVA and Dunnett's test.Results (1) Western blot test:the expression levels of Ang-2 and HIF-2α proteins in HUVECs under hypoxia conditions at 0,2,4,8,12,16,20 hours were 0.110 ±0.011,0.120 ±0.020,0.210 ±0.070,0.410 ±0.100,0.520 ± 0.090,0.790±0.130 1.010 ±0.220 and 0.180 ±0.090,0.410 ±0.070,0.470 ±0.110,0.470 ±0.070,0.580 ± 0.120,0.690 ± 0.140,0.920 ± 0.130,respectively,and which were increased after culturing under hypoxia conditions and had an ascending tendency as the hypoxia time extended,with statistically significant differences (F =403.550,3 265.587,P ＜ 0.05).The expression levels of Ang-2 and HIF-2α proteins in the control group,shRNA control group and HIF-2α shRNA group were 1.030 ±0.180,1.070 ±0.120,0.210 ± 0.070,and 0.940 ± 0.110,0.930 ± 0.190,0.170 ± 0.021,respectively,showing statistically significant differences (F =290.242,26.688,P ＜ 0.05).(2) The results of ELISA:the expression levels of Ang-2 in the control group,shRNA control group and HIF-2α shRNA group were (433.2 ±9.7)ng/L,(438.3 ± 2.6)ng/L,(114.6 ± 4.2) ng/L,with a statistically significant difference (F =2 642.180,P ＜ 0.05).(3) The results of tube formation experiments:the number of endothelial cell tubes in the control group,shRNA control group,HIF-2α shRNA group and HIF-2α ± rh-Ang-2 group were 48.3 ± 2.5,47.4 ± 3.1,19.7 ± 1.5 and 38.3 ± 2.1,respectively,with a statistically significant difference (F =148.196,P ＜ 0.05).(4) The results of Transwell method:① the number of HUVECs migration in the control group,shRNA control group,HIF-2α shRNA group and HIF-2α + rh-Ang-2 group were 140.3-± 3.5,142.7 ± 2.1,42.7 ± 3.1 and 78.1 ± 4.2,respectively,showing a statistically significant differences (F =212.205,P ＜ 0.05).②The results of Transwell method:the number of SMMC-7721 cells migration after intervening using four different supernatant in the control group,shRNA control group,HIF-2α shRNA group and HIF-2α ± rh-Ang-2 group were 106.7 ± 5.5,102.7 ± 6.6,63.0 ± 3.3 and 96.7 ± 2.1,respectively,showing a statistically significant difference (F =55.122,P ＜ 0.05).Conclusion HIF-2a could not only affect HUVECs formation but also promote SMMC-7721 cells migration via regulating Ang-2 expression.

Objective To investigate the function characteristics of CD4 T cell converted double negative T cell and provide a basis for further insight into the characteristics of mouse converted double negative T cell.Methods The gene expression profile was analyzed by transcriptome sequencing and protein mass spectrometry.The expression of cell active marker CD44,CD69 and OX40 was investigated by flow cytometry and the cytotoxicity of mouse double negative T cell was verified by CFSE staining.Results Mouse CD4 T cell converted double negative T cell expressed cell phenotype that differed from other mature CI4 T cells.Mouse converted double negative T cell expressed high level of active marker of CD44,CD69 and OX40.Cytotoxicity of PrfO DN T was significantly reduced.Conclusions Mouse CD4 T cell converted double negative T cell has distinguishing cell phenotypes,that are not identical to other mature CD4 T cells.Mouse double negative T cell overexpresses cell activation marker and cytotoxic cytokines.The immune suppressive function of mouse double negative T cell is mainly dependent on perforin pathway.

Hepatocellular carcinoma (HCC) owns the high morbidity and mortality rates. Surgical resection is still the major pathway for the longer survival of HCC patients. Postoperative recurrence and metastasis have become the key impairment of prognosis of HCC patients. The relationship between tumor recurrence and surgical manner underwent by HCC patients is complicated and multiple factors are included. When the liver tumor was pressured during operation, tumor cells could be squeezed into blood flow via the broken vessels, which resulted in tumor metastasis. Besides, ischemia-reperfusion injury induced by Pringle maneuver during the liver blood blockade resulted in the immune destruction of liver and induced tumor recurrence.The destruction of physical barriers consisted of interstitial cells and normal liver cells was also a key factor for tumor recurrence. This paper summarizes the possible relationship between postoperative recurrence and surgical manner in HCC patients to provide the preventive suggestions for the postoperative recurrence of HCC patients.

Hepatocellular carcinoma (HCC) owns the high morbidity and mortality rates. Surgical resection is still the major pathway for the longer survival of HCC patients. Postoperative recurrence and metastasis have become the key impairment of prognosis of HCC patients. The relationship between tumor recurrence and surgical manner underwent by HCC patients is complicated and multiple factors are included. When the liver tumor was pressured during operation, tumor cells could be squeezed into blood flow via the broken vessels, which resulted in tumor metastasis. Besides, ischemia-reperfusion injury induced by Pringle maneuver during the liver blood blockade resulted in the immune destruction of liver and induced tumor recurrence.The destruction of physical barriers consisted of interstitial cells and normal liver cells was also a key factor for tumor recurrence. This paper summarizes the possible relationship between postoperative recurrence and surgical manner in HCC patients to provide the preventive suggestions for the postoperative recurrence of HCC patients.

Objective To investigate the biomechanical effects of the backside design of tibial trays on the bone-prosthesis fixation interfaces in unicompartmental knee arthroplasty(UKA).Methods Finite element models of medial knee arthroplasty were constructed using a fixed UKA prosthesis.The knee joint load and joint motion under walking motion were considered as boundary conditions,and the differences in tibial von Mises stress,contact stress,and micromotion of the bone-prosthesis fixation interface of the UKA tibial trays with big keel,small keel,two-peg with fin,three-oblique peg,and three-upright peg types were compared.Results At the maximum medical knee force moment,compared to the two-peg with fin type,the tibial von Mises stress,contact stress,and micromotion of the bone-prosthesis fixation interface decreased by 8%and 15.9%and increased by 9.9%for the big keel type;decreased by 12.3%and increased by 7.5%and 0.9%for the small keel type;decreased by 10%,10.5%,and increased by 1.2%for the three-oblique peg type;and decreased by 7.7%,14.7%,and 1.6%for the three-upright peg type,respectively.However,the maximum micromotion of the bone-prosthesis fixation interface occurred at 21%of the gait cycle.Compared to the two-peg with fin type,the micromotion of the bone-prosthesis fixation interface increased by 11.6%for the big keel type,increased by 1.6%for the small keel type,decreased by 0.4%for the three-oblique peg type,and decreased by 2.3%for the three-upright peg type.Conclusions To improve the long-term fixation effects of tibial prostheses,it is recommended to focus on a two-upright peg with fin or small keel designs when UKA tibial trays are designed,which can effectively balance the stress transfer and interface micromotion,thereby ensuring prosthesis stability and reducing the risk of aseptic loosening.

Sinapic acid (SA) is a phenolic acid that is widely distributed in fruits and vegetables, which has various bioactivities, such as antidiabetic, anticancer and anti-inflammatory functions. Over-activated microglial is involved in the development progress of neurodegenerative diseases, such as Parkinson’s disease and Alzheimer’s disease. The objective of this study was to investigate the effect of SA in microglia neuroinflammation models. Our results demonstrated that SA inhibited secretion of the nitric oxide (NO) and interleukin (IL)-6, reduced the expression of inducible nitric oxide synthase (iNOS) and enhanced the release of IL-10 in a dose-dependent manner. Besides, our further investigation revealed that SA attenuated the phosphorylation of AKT and MAPK cascades in LPS-induced microglia. Consistently, oral administration of SA in mouse regulated the production of inflammationrelated cytokines and also suppressed the phosphorylation of MAPK cascades and AKT in the mouse cerebral cortex. These results suggested that SA may be a possible therapy candidate for anti-inflammatory activity by targeting the AKT/MAPK signaling pathway.

An estimate of about 50% of new liver cancer cases worldwide occur in China every year.Surgical resection is still the major treatment choice for longer survival of patients with hepatocellular carcinoma. Blocking hepatic blood flow and reducing intraoperative bleeding ensure the success of the operation. Anatomic separation of hepatic hilar region is the precondition of hepatic inflow occlusion. The hepatic hilar plate system involves a thick layer of connective tissue covering the hepatic inflow ducts of hepatic hilar region. The descending part of hilar plate assists in reducing the anatomical difficulty of the hepatic hilar region. The "forth porta hepatis" that is hidden in the hepatic hilar plate system involves the accumulation area of "short hepatic portal veins" .The communicating branch vessels between the hepatic inflow vessels form the anatomical basis in reducing the indocyanine green fluorescence stain effect.The relatively fixed position of the hepatic portal plate is considered as a positioning marker for accurate liver resection. The intrahepatic Glisson sheath is connected with thick connective tissue of the hepatic portal panel system, and is regarded as the physical barrier in limiting the proliferation and hypertrophy of hepatocytes and continuation of hepatic portal panel system in the liver.This paper summarizes the anatomy and application of hepatic hilar plate system during hepatobiliary surgery.

Background:The musk glands of adult male Chinese forest musk deer(Moschus berezovskii Flerov,1929)(FMD),which are considered as special skin glands,secrete a mixture of sebum,lipids,and proteins into the musk pod.Together,these components form musk,which plays an important role in attracting females during the breeding season.However,the relationship between the musk glands and skin of Chinese FMD remains undiscovered.Here,the musk gland and skin of Chinese FMD were examined using histological analysis and RNA sequencing(RNA-seq),and the expression of key regulatory genes was evaluated to determine whether the musk gland is derived from the skin.Methods:A comparative analysis of musk gland anatomy between juvenile and adult Chinese FMD was conducted.Then,based on the anatomical structure of the musk gland,skin tissues from the abdomen and back as well as musk gland tissues were obtained from three juvenile FMD.These tissues were used for RNA-seq,hematoxylin-eosin(HE)staining,immunohistochemistry(IHC),western blot(WB),and quantitative real-time polymerase chain reaction(qRT-PCR)experiments.Results:Anatomical analysis showed that only adult male FMD had a complete glandular organ and musk pod,while juvenile FMD did not have any well-developed musk pods.Transcriptomic data revealed that 88.24%of genes were co-expressed in the skin and musk gland tissues.Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway analysis found that the genes co-expressed in the abdomen skin,back skin,and musk gland were enriched in biological development,endocrine system,lipid metabolism,and other pathways.Gene Ontology(GO)enrichment analysis indicated that the genes expressed in these tissues were enriched in biological processes such as multicellular development and cell division.Moreover,the Metascape predictive analysis tool demonstrated that genes expressed in musk glands were skin tissue-specific.qRT-PCR and WB revealed that sex-determining region Y-box protein 9(Sox9),Caveolin-1(Cav-1),and androgen receptor(AR)were expressed in all three tissues,although the expression levels differed among the tissues.According to the IHC results,Sox9 and AR were expressed in the nuclei of sebaceous gland,hair follicle,and musk gland cells,whereas Cav-1 was expressed in the cell membrane.Conclusions:The musk gland of Chinese FMD may be a derivative of skin tissue,and Sox9,Cav-1,and AR may play significant roles in musk gland development.