Objective To study the expression of p35 and p25 in rat after focal cerebral contusion and to provide experimental data for estimating brain injury time. Methods Fifty adult male SD rats were randomly divided into 0 h, 6 h, 12 h, 24 h, 3 d, 5 d, 7 d, 10 d after focal cerebral contusion, control and sham-operated groups (5 rats each group). The focal cerebral contusion rat model was established. The expression of p35 and p25 protein of the damage peripheral zone in brain were detected by HEstain-ing, immunohistochemistry and western blotting at different injury time. Results Alarge number of p35 protein and a small amount of p25 protein were expressed in control group and sham-operated group. After focal cerebral contusion, p35 presented unimodal change with time and p25 presented bimodal changes with time. Conclusion Expression of p35 and p25 showed different regularity with good time correlation, which could help to estimate the brain injury time.

Objective To evaluate the efficacy and security of metformin in the treatment of endocrine resistance and postmenopausal hormone receptor positive advanced breast cancer. Methods 60 cases of postmenopausal HR+advanced breast cancer whose first-line or second-line endocrine therapy failed were randomly divided into study group(n=30),treated with metformin combined with AI and control group(n=30),treated with placebo combined with AI. Standard RECIST guidelines were used to evaluate the clinical response. The objective response rate(ORR),clinical benefit rate(CBR),progression-free survival,and adverse reactions of two groups were compared. Results The ORR of two groups were 16.7%and 10%respectively and the difference was not statistically significant(P > 0.05). But CBR in study group was significantly higher than that in control group (63.3%vs 36.7%),and the difference was statistically significant(P<0.05). The median PFS in study group was slightly longer than that in control group (3.7 m vs 4.2 m),but there was no statistical difference. Multivariate regression analysis showed that PFS was only associated with the previous endocrine therapy. No serious adverse reactions occurred in two groups. Conclusion Metformin is expected to improve secondary endocrine resistance in breast cancer,but large prospective clinical studies are needed to confirm it.

Objective To explore the fluorescent distribution, and the relation between fluorescent intensity and time after injecting rear-thigh muscles with DiI. Methods Sixty-five new born mice were equally and randomly divided into 13 groups. Fluorescent distribution and intensity were investigated at 2, 4, 6, 8, 12, 24 h, and 2, 4, 7, 14, 28 d and 2, 3 months after injecting the left posterior limb rear thigh muscles with 1.5 ?l 4 mg/ml DiI for one mouse. Results The labeling neurons were scattered from L2 level to S2 level and associated dorsal root ganglion (DRG), but the most were located at L4 to L6 section. The faint red fluorescence neurons were observed at dorsal root ganglion and cornu anterius medullae spinalis 6 h post-injection. The labeling neurons increased up to the 4th day. The fluorescent intensity enhanced gradually from 6 h to 24 h, then kept the intensity for 3 months. Conclusion It is a quick, precise, persistent method to trance and label the dorsal root ganglions sensory neuron and spinal motoneuron by injecting rear-thigh muscles with DiI, and the rate of labeling neurons can be improved by prolonging the tracing time properly. It is also provide basic data for clinical or experimental neuron label and location.

Objective This study aims to evaluate the changes of Cdk5 expression at the time of 3 hours to 10 days after moderate brain injury by blunt force impact in a rat model,and to demonstrate its forensic significance.Methods To establish a rat model of blunt focal brain contusion,and to observe the changes of Cdk5 expression in brain tissue at different timepoints after brain injury by immunohistochemistry and Western blot.Results A low expression level of Cdk5 was observed in the brain tissue of both normal and sham control groups.The expression of Cdk5 increased after 3 and 6 hours,remarkably increased at 12 hours,and reached the maximal level at 24 hours after focal brain injury.The Cdk5 level gradually decreased 3 days,5 days,7 days,and 10 days and reached the normal level 7 and 10 days after the injury,with no statistical difference (P＞0.05) compared with the normal and sham control groups.Conclusion The expression of Cdk5 increased in the peripheral area of contusion tissue after blunt brain injury in rats,showing single peak change,and dropped to normal level with the time extension.The change of Cdk5 expression may provide a new reference index for the prediction of early brain contusion.

Objective To evaluate the effect of endoplasmic reticulum stress in trophocytes,in patients with intrahepatic cholestasis of pregnancy (ICP).Methods Sixty-one pregnant women who were hospitalized in Women's Hospital,School of Medicine,Zhejiang University from January to December 2015 were recruited.Thirty-one women who were diagnosed as ICP were defined as the ICP group and 30 healthy pregnant women were defined as the control group.The localization and expression intensity of glucose regulated protein 78 (GRP-78) in placental tissues were detected by immunohistochemistry technique.Electronic microscope was used to observe ultra-microstructure change of the endoplasmic reticulum in trophocytes and cell line Swan71.Reverse transcription (RT)-PCR and western blot were used to investigate the expression of GRP-78 mRNA and protein in Swan 71 cell.Results (1) GRP-78 protein was mainly expressed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts.The protein expression of GRP-78 in placentas of the ICP group (13.2±2.4) was significantly higher than that in the control group (7.8±1.3,P＜0.01).(2) The volume of endoplasmie reticulum did not increase and the microvilli developed well,with no swelling and no expansion of endoplasmic reticulum in the control group.In the ICP group,microvilli injury,endoplasmic reticulum edema were found;the volume of endoplasmic reticulum increased,with dilation,vacuolation and significant degranulation.After treated with 100 μmol/L cholyglycine for 24 hours,universal dilatation of the endoplasmic reticulum were seen in the Swan71 cells.(3) In Swan71 cells,cholylglycine displayed a concentration-dependent up-regulation on the expression of GRP-78.The expressions of GRP-78 mRNA in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.17,2.17±0.16,5.47±0.36,5.65 ± 0.82,respectively.The expression of GRP-78 protein in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.04,1.17±0.15,1.33±0.13,1.73±0.13,respectively.The expression of GRP-78 mRNA and protein in 100 and 50 μ mol/L cholylglycine experimental group were significantly higher than 0 μmol/L (all P＜0.01).Conclusion The obvious expansion of endoplasmic reticulum and the increased expression of GRP-78 in trophocytes indicated that endoplasmic reticulum stress of trophocytes may be involved in the pathogenesis of ICP.

The liver has a strong regenerative capacity that ensures patient recovery after hepatectomy and liver transplantation.The portal system plays a crucial role in the dual blood supply to the liver,making it a significant factor in hepatic function.Several surgical strategies,such as portal vein ligation,associating liver partition and portal vein ligation for staged hepatectomy,and dual vein embolization,have high-lighted the portal system's importance in liver regeneration.Following hepatectomy or liver trans-plantation,the hemodynamic properties of the portal system change dramatically,triggering regeneration via shear stress and the induction of hypoxia.However,excessive portal hyperperfusion can harm the liver and negatively affect patient outcomes.Furthermore,as the importance of the gut-liver axis has gradually been revealed,the effect of metabolites and cytokines from gut microbes carried by portal blood on liver regeneration has been acknowledged.From these perspectives,this review outlines the molecular mechanisms of the portal system's role in liver regeneration and summarizes therapeutic strategies based on the portal system intervention to promote liver regeneration.

[Objective]To explore the phenomenon and regularity of acupoint sensitization for major depressive disorder(MDD)based on bibliometric methodology and data mining technology,and to provide scientific ideas for the clinical diagnosis and treatment of MDD.[Methods]Eight electronic databases and one ancient book were comprehensively searched and available literature was retrieved.Bibliometric methodology and data mining technology were applied to summarize the phenomenon and regularity of acupoint sensitization for MDD,and to explore the related theoretical mechanisms.[Results]A total of 29 literature related to acupoint sensitization of MDD was included.Acupoint sensitization of MDD involved in multiple forms,including heat sensitization,chemical sensitization,pain sensitization,and so on,and the testing means were diverse.The most common sensitized acupoint was Baihui(GV20).Among these acupoints,sensitization often simultaneously occurred in Baihui(GV20)and Neiguan(PC7),Baihui(GV20)and Taichong(LR3).The acupoints and sensitization could be divided into 4 effective clusters.The meridian involved with high sensitization frequency was governor vessel.Sensitization mainly occurred in the head-face-neck area.[Conclusion]The sensitization phenomenon of MDD has many types,However,there are few related studies at present.The phenomenon of acupoint sensitization involving acupoint,meridians,and location are relatively broad.There might be a close relationship between high-frequency sensitized acupoints and common acupoints for clinical treatment,which may be related to the effect of different acupoints and the etiology and pathogenesis of MDD.

Objective:To observe any effect of electroacupuncture (EA) on the expression of phosphorylated extracellular signal-regulated protein kinase (p-ERK1/2) and phosphorylated cyclic adenosine monophosphate response element binding protein (p-CREB) in the spinal dorsal horns of diabetics experiencing neuropathic pain.Methods:Eight rats were randomly selected from 30 healthy male Sprague-Dawley rats as the normal group (N), and the remaining twenty-two rats were treated with a single high-dose intraperitoneal injection of streptozotocin (STZ) to establish a neuropathic pain model. The rats modeled successfully were randomly divided into a model group (M, n=8) and an EA group ( n=8). In the EA group, electroacupuncture was applied at the bilateral Hou san li and Kunlun acupoints starting on the 15th day after the STZ injection. The daily sessions lasted 30 minutes for 1 week. Body weight (BW), fasting blood glucose (FBG) and paw withdrawal latency (PWL) were observed before the STZ injection and on the 7th, 14th, and 21st days afterward. The expression of p-ERK1/2 and p-CREB in the dorsal horns of the rats′ spinal cords was detected using western blotting. The count of p-CREB-positive cells in the dorsal horns and their co-localization with neurons was detected using immunofluorescence. Results:In comparison with the N group, the average BW of the M group on the 7th, 14th and 21st days after the STZ injection was significantly lower, while the average FBG was significantly higher. There was no significant difference between the M and N groups in the average PWL on the 7th day after the STZ injection, but it had decreased significantly in the M group on the 14th and 21st days. Compared with the M group, the average PWL of the EA group was significantly longer on the 21st day after the injection. The expression of p-ERK1/2 and p-CREB protein in the spines of the M group was significantly higher than in the N group. p-CREB positive cells were more numerous in the M group compared with the N group, while in the EA group they were fewer. P-CREB was co-located with neurons in the spinal dorsal horn.Conclusion:EA can alleviate neuropathic pain effectively, perhaps by inhibiting the expression of p-ERK1/2 and p-CREB in the dorsal horns of the spinal cord.

Skin ulcers are the most prevalent consequence of diabetes mellitus, and people with diabetic ulcers have a substantially greater death risk than those who do not have ulcers. Herbal medications have gained wide concern in recent years due to their multi-component, multi-target, and multi-pathway synergistic therapeutic effects. Clinical trials have demonstrated the safety and efficacy of herbal treatments in diabetic refractory ulcers. To systematically evaluate the healing effect of herbs on diabetic wounds, a literature search was conducted, the mechanism of action of 15 herbal extracts in promoting diabetic wound healing were reviewed, and the classification based on traditional Chinese medicine theory was discussed, which could provide a reference for the precise treatment and exploitation of herbal medicines for diabetic ulcers.

Objective: To investigate the changes of microorganisms and metabolites in joint effusion of patients with Rheumatoid arthritis(RA), Osteoarthritis(OA) and Urarthritis(UA). To provide new ideas for the study of the effect of microbiota on the pathogenesis of arthritis. Methods: Joint effusion samples were collected from 20 patients with RA, 20 patients with OA, and 20 patients with UA. 16S rRNA gene sequencing and untargeted ultra-high performance Liquid chromatography-mass spectrometry(LC-MS) were used to explore the differences in microorganisms and metabolites among the three groups. Pearson correlation analysis was used to detect the correlation between effusion microbiota and metabolites. Results: There were differences in microbial diversity and microbiota composition among the three groups. Combined with VIP>1 from OPLS-DA andP<0.05 from two-tailed Students t-test, 45 differential metabolites(Between RA and OA groups), 38 differential metabolites(Between UA and OA groups) and 16 differential metabolites(Between RA and UA groups), were identified. GO analysis and KEGG pathway analysis showed that the differential metabolic pathways among the three groups were mainly concentrated in citric acid cycle(TCA cycle), nucleotide metabolism, amino acid metabolism and glycolysis pathway. Correlation analysis of joint effusion microbiota and metabolites suggested that bacteria enriched in the three groups of joint effusion, such asPrevotella,Clostridium ruminosus,Prevotellaceae＿UCG-001, were related to many key metabolites such as lysozyme, uric acid, glucose, and L-glutamine. Conclusion This study shows that there are a variety of bacterial flora in joint cavity effusion of RA, OA, and UA patients, and the differential metabolites produced by them are involved in the pathogenesis of the three types of arthritis by affecting a variety of metabolic pathways.