Neijing was a early medical classic existing in China and had a deep influence on the formation of traditional Chinese medicine. Besides, Neijing was also an ancient philosophic works, in which the severe view of heaven was recorded, laying a sound basis for the development of TCM. It would have great significance for health cultivation of patients with chronic liver diseases to analyze the theory on relation between human and nature recorded in Neijing and to absorb its rational contents.

Objective To study the relationships among leptin, leptin receptor (OB-R) gene in arcuate nucleus and type 2 diabetes mellitus, and the effect of acupuncture on them. Methods Obese rats due to diet were injected in abdomen with small dosage streptozotocin to induce type 2 diabetes mellitus model, and were divided into acupuncture, glibenclamide and model group. Normal rats were in control. After treatment for 4 weeks, fasting blood sugar (FBS), fasting insulin (FINS), leptin, OB-R mRNA expression in arcuate nucleus were evaluated. Height, weight were measured and Lee's index were calculated before and after treatment. Results The FBS and FINS were higher in model group ［(15.79±1.87) mmol/L, (37.20±3.92) mIU/L］ than in control ［(5.16±0.13) mmol/L, (23.57±1.63) mIU/L］, but decreased in acupuncture group ［(5.46±0.20) mmol/L, (20.87±1.80) mIU/L］ and glibenclamide group ［(5.26±0.13) mmol/L, (28.10±1.86) mIU/L］. Leptin was lower in model ［(0.95±0.15) ng/ml］ than in control ［(1.32±0.15) ng/ml］, but was higher in glibenclamide group ［(1.83±0.23) ng/ml］ than in control. Weight and Lee's index decreased in model, acupuncture and glibenclamide groups after treatment. OB-R mRNA expression in arcuate nucleus in model wasn't different from control, acupuncture or glibenclamide group, nor between acupuncture and control, but was lower in glibenclamide group than in control. Conclusion Leptin decreased in type 2 diabetes mellitus rats when they became thin. Leptin resistance appears when treated with glibenclamide. Acupuncture may improve the OB-R mRNA expression in arcuate nucleus to some extent.

Objective:To study the relationship between hepatic tissue insulin receptor substrate 2(IRS2) gene and type 2 diabetes,and the regulation of acupuncture on IRS2 gene expression of hepatic tissue in type 2 diabetes mellitus(T2DM) rats.Methods:Obese rats due to diet were injected in abdomen with small dose streptozotocin(STZ) to induce T2DM.Then these T2DM rats were stochastically divided into acupuncture group,Glibenclamide group and model group.After 4 weeks treatment,fasting blood sugar(FBS) was examined with active blood sugar meter,fasting insulin(FINS) was examined by radioimmunoassav,hepatic tissue IRS2mRNA was evaluated in real-time RT PCR,and normal rats were in control.Results:Serum levels of FBS(15.79?1.87) and FINS(37.20?3.92) in model group were higher than those in normal group(5.16?0.13,23.57?1.63),serum levels of FBS(5.46?0.20) and FINS(20.87?1.80) in acupuncture group were obviously lower than those in model group,which were similar to normal group.FBS(5.26?0.13) reduced obviously and serum FINS(28.10?1.86) reduced in some degree in Glibenclamide group compared with model.Relative contents of IRS2mRNA in model group were 2.19 folds compared with that in normal group.And in Glyburide group it was 4.59 folds compared with that in normal group.In acupuncture group it was 4.04% of that in the normal group.Conclusion:Abnormal hepatic tissue IRS2 mRNA expression may be one of pathogenesis of T2DM.Acupuncture can obviously depress hepatic tissue IRS2 mRNA expression,but Glibenclamide can not.

BACKGROUND:There is no effective treatment for muscle atrophy caused by peripheral nerve damage. Skeletal muscle cel s, a structural unit of muscle contraction, can be used for studies on muscle atrophy when cultured in vitro.
OBJECTIVE:To investigate the promotion effect of neuron-secreted factors on the growth of skeletal muscle cel s in vitro.
METHODS:Skeletal muscle cel s primary cultured in vitro were divided into two groups:experimental group with neuron-secreted factors, and control group with common culture medium, respectively. Afterwards, the number of skeletal muscle cel s and expression level of alpha actin were detected by hematoxylin-eosin staining and immunohistochemical staining, respectively.
RESULTS AND CONCLUSION:The number of skeletal muscle cel s and expression level of alpha actin in the experimental group were significantly higher than those in the control group (P<0.05). In conclusion, neuron-secreted factors have the ability of promoting the growth of skeletal muscle cel s and may be helpful for denervated muscle atrophy.

AIM: To investigate the association of osteoprotegerin ( OPG) gene single nucleotide polymor-phisms (SNPs), 163A/G (rs3102735) and 245T/G (rs3134069), with susceptibility to rheumatoid arthritis (RA) in Chinese Han population .METHODS:A total of 205 patients with RA and 171 healthy control subjects were enrolled into this study.Genotyping was performed by polymerase chain reaction-based restriction fragment length polymorphism and subsequently confirmed by DNA sequencing .Odds ratio ( OR) and 95%confidence intervals ( CI) were calculated for the risk genotypes and alleles .RESULTS: OPG gene polymorphisms 163A/G and 245T/G were conformed to the Hardy-Weinberg equilibrium .The statistical differences in the genotypes of AA , AG and GG at 163A/G locus were found in RA and controls.The G allele was associated with an increased risk of RA , with OR of 1.219 (95%CI:1.066~2.339).No significant difference was observed between RA group and control group with respect to genotypic and allelic frequencies of OPG gene 245T/G (P>0.05).CONCLUSION:The OPG gene 163A/G SNP may be associated with RA susceptibility , and G allele may be the risk factor for developing RA .

Objective To observe the functional connectivity (FC) pattern linking the hippocampus with the rest of the brain in ischemic stroke patients with cognitive dysfunction,especially the default mode network (DMN).Methods Resting-state functional connectivity magnetic resonance imaging (fMRI) was performed on 15 ischemic stroke patients with cognitive dysfunction (the patient group) and 10 normal elderly controls.The bilateral hippocampus was the region of interest.Correlation analyses yielded a mapping of cerebral functional connectivity activation for both groups.Results Compared with the healthy controls,the patient group showed weakened functional connectivity between the hippocampus and other regions including the cingulate gyrus,the superior,middle and inferior frontal lobes,the inferior parietal lobule and the superior temporal gyrus.But there was enhanced functional connectivity with the cerebellar posterior lobe,the occipital lobe,the medial temporal lobe,the precuneus and the calcarine.The results were significantly different between the two groups.Conclusion The functional connectivity pattern of the hippocampus is impaired in stroke patients with cognitive dysfunction.Reduced functional connectivity between brain regions may be one cause of cognitive dysfunction after stroke,and enhanced functional connectivity may be an appropriate compensatory mechanism.

Objective To compare the effect of transplant islets between the subcutaneous inguinal white adipose tissues and renal capsule in the treatment of type 1 diabetes mellitus in mouse models. Methods The mice with type 1 diabetes mellitus undergoing islet transplantation were divided into the white adipose group (n=10) and renal capsule group (n=10). The islets were isolated, purified and transplanted to the subcutaneous white adipose tissues of inguinal region and renal capsule. The random blood glucose level and glucose tolerance function of the recipient mice in two groups were continuously monitored after operation. Islet grafts of the surviving recipient mice were harvested at postoperative 100 d for histopathological examination. Results In the white adipose group, the blood glucose levels of 6 recipient mice were restored to normal at 1 month after transplantation, whereas the blood glucose levels of the other 4 recipient mice were high, which died before the end of monitoring. In the renal capsule group, the blood glucose levels of 10 recipient mice returned to normal within 10 d after transplantation. Islet grafts of the recipient mice in two groups could lower the blood glucose levels, whereas the islet grafts in the white adipose group required a longer time to exert the effect. The glucose tolerance function of the mice in the renal capsule group was significantly better than that of those in white adipose group (P < 0.05). Histopathological examination demonstrated that the insulin of the islet grafts was normally expressed in two groups. Conclusions The islets transplanted into the subcutaneous white adipose tissues of inguinal region can play an effective role in regulating the changes of blood glucose level. Although the blood glucose-lowering function is slightly weaker than that of the islets graft in the renal capsule, it has multiple advantages resembling the ideal islet transplantation sites, which is a promising replacement site for islet transplantation.

Objective:To investigate effects of the autophagy inducer rapamycin and autophagy inhibitor 3-methyladenine on viral structures and biosynthesis of functional proteins in dorsal root ganglia in a guinea pig model of varicella-zoster virus (VZV) infection, and to explore their possible mechanisms.Methods:VZV was cultured and proliferated in human embryonic lung fibroblasts (HELFs) , and peripheral blood mononuclear cells (PBMCs) were isolated from guinea pigs. VZV-HELFs and PBMCs were co-cultured for 18-20 hours, and VZV-PBMCs were collected by centrifugation. Thirty-two guinea pigs were randomly and equally divided into 4 groups (8 mice in each group) : blank control group was injected with autologous PBMCs via the medial canthal venous plexus; autophagy inhibition group, autophagy induction group, and VZV infection group were intraperitoneally injected with 3 mg/kg 3-methyladenine solution, 0.5 mg/kg rapamycin solution, and the same volume of 0.9% NaCl solution respectively, followed 2 hours later by injections with 50 μl of VZV-PBMCs via the medial canthal venous plexus. Fourteen days later, the guinea pigs in each group were sacrificed, and dorsal root ganglion tissues were collected. The transmission electron microscope was used to observe the morphology of virus particles, as well as the morphology and number of autophagic vesicles, Western blot analysis was performed to determine the expression of VZV nucleocapsid protein (NCP) , immediate-early protein 62 (IE62) , and autophagy-related proteins Beclin-1 and p62, and immunohistochemical study to determine the expression of anti-VZV antibodies in VZV-infected dorsal root ganglia. Statistical analysis was carried out by using two-independent-sample t test, one-way analysis of variance, least significant difference- t test or Kruskal-Wallis H test. Results:Nucleocapsid-containing virions and scattered autophagosomes were seen in the dorsal root ganglia in the VZV infection group under the transmission electron microscope. The number of autophagic vesicles significantly differed among the blank control group, VZV infection group, autophagy induction group and autophagy inhibition group ( M[ Q1, Q3]: 0, 5[4, 6], 7[5, 9], 0, respectively; H = 135.60, P < 0.01) , and was significantly higher in the VZV infection group than in the blank control group and autophagy inhibition group (both P < 0.05) , as well as in the autophagy induction group than in the autophagy inhibition group ( P<0.05) , but there was no significant difference between the VZV infection group and autophagy induction group ( P>0.05) . Western blot analysis showed that the expression level of IE62 protein was significantly higher in the VZV infection group (1.49 ± 0.06) than in the blank control group (0.50 ± 0.09, t = 9.17, P < 0.05) ; the expression of anti-VZV antibodies was significantly lower in the autophagy inhibition group than in the autophagy induction group and VZV infection group ( t = 9.24, 7.78, respectively, both P < 0.01) , while there was no significant difference between the autophagy induction group and VZV infection group ( P > 0.05) . Conclusion:Autophagy occurred in the dorsal root ganglia of guinea pigs after VZV infection; the inhibition of autophagy could affect the structure of VZV and decrease the expression of VZV functional proteins in the dorsal root ganglia of guinea pigs.

To establish a platform to monitor the immune rejection after abdominal aortic patch suture in a xenotransplantation model.Methods The carotid was excised from wild-type Bama pigs,cut into 2.5 cmx 1.0 cm pieces in shuttle shape and subsequently sutured to the abdominal aorta of cynomolgus monkeys.No immunosuppressive agent was administered.General conditions of the recipient monkeys were observed.The morphological changes of the graft artery were assessed by pathological examination at postoperative 1 year.Before and 7,14,28 and 49 d after surgery,the blood samples were collected from the recipient monkeys.The serum levels of IgM and IgG antibodies were quantitatively measured by the red blood cell and peripheral blood mononuclear cell (PBMC) from Bama pigs.The quantity of lymphocytes in the recipient monkeys was detected by routine blood test and flow cytometry.Results All 3 monkeys undergoing transplantation survived well.At postoperative 1 year,the lateral tissues of the vascular wall at the artery graft were seen in dark red color.Hematoxylin-eosin (HE) staining revealed a large quantity of red blood cell and platelet deposition,accompanied with lymphocyte infiltration.Using porcine red blood cell and PBMC as target cells,the serum levels of anti-pig IgM and IgG antibodies peaked at postoperative 28 d,and slightly declined at postoperative 49 d.The quantity of lymphocytes and T cell subset also peaked at postoperative 28 d and began to decrease at postoperative 49 d.Conclusions Artery patch suture is a simple and reliable xenotransplantation model.The recipients can maintain normal physiological state without the use of immunosuppressive agents.The grafts can effectively activate the immune system of the recipients,induce the production of anti-pig antibodies and provoke cellular immune rejection.Therefore,this model can be utilized to monitor the immune rejection throughout the xenotransplantation process.

Objective:To explore genes related to costimulatory molecule related to the prognosis of bladder cancer,and to construct and evaluate prognosis model based on costimulatory molecule-based signature(CMS).Methods:Gene expression matrix and clinical information of bladder cancer patients were downloaded from TCGA database and GEO database(GSE31684),and costimulatory molecule-related genes were retrieved from the literature.The univariate and multivariate Cox analysis were used to screened prognostic-related genes and constructed prognostic model.Forecast accuracy of model was verified in TCGA training group,TCGA validation data group and GEO group by Kaplan-Meier survival analysis and receiver operating characteristic curve(ROC).Considering risk score and clinical characteristics,we constructed a nomogram and evaluated its performance by consistency analysis and ROC.CIBERSORT algorithm was used to analyze immune cell composition of tumor microenvironment infiltration,and gene set enrichment analysis(GSEA)was performed to explore the potential mechanism.Results:Four prognostic-related CMSs were found:TNFRSF14,CD276,ICOS and TMIGD2,of which three were included in the risk score construction.Multivariate Cox regression results showed that the risk score based on CMS was an independent prognostic factor for bladder cancer patients.Consistency analysis and ROC results showed that the nomogram had ideal prognosis prediction accuracy.Immune infiltration analysis showed that the high risk group was likely to be in immunosuppressive state.GSEA results suggested that genes in high risk group were enriched in extracel-lular matrix(ECM)receptors interaction,cell cycle and other pathways.Conclusion:TNFRSF14,CD276 and ICOS may be potential prognostic biomarkers for bladder cancer patients.CMS-based risk score and nomogram could contribute to early prognosis and choice of personalized treatment.