Objective To investigate the expressions of OCT4 and P27kip1 and their correlations in renal hyaline cell carcinoma. Methods A total of 24 samples of renal hyaline cell carcinoma were detected by immunohistochemistry. The rate of cells with positive labelling of OCT4 and P27kip1 and their intra cellular distribution were observed in renal hyaline cell carcinoma. The expression levels of OCT4 and P27kip1 were compared between different gender, age (<60y and≥60y) and cancer cell metastasis groups. Results The rates of cells with positive OCT4 and P27kip1 expressions were 66.7%and 75% in renal hyaline cell carcinoma respectively. The ratio of cells with high, middle and low expression of OCT 4 were 33.3%, 20.5%and 12.5%in renal hyaline cell carcinoma tissues. OCT4 was mainly expressed in cytoplasm and nuclei with a few was expressed in the cell membrane. The ratio of cells with high, middle and low expression of P27kip1 were 12.5%, 25%and 37.5%in renal hyaline cell carcinoma tissues respectively. The positive staining of P27kip1 was in the cytoplasm and cell membrane, and a small number of nuclei were expressed. There were no significant differences in OCT4 and P27kip1 between different age and gender groups. There were significant differences in OCT4 and P27kip1 expressions between patients with metastasis and patients without metastasis (P < 0.05). A negative correlation was found between OCT4 and P27kip1 expres?sions in renal hyaline cell carcinoma (rs=-0.408, P<0.05). Conclusion The expression of OCT4 is negatively correlated with the expression of P27kip1. Inhibiting expressions of OCT4 or P27kip1, especially knocking down P27kip1 in cytoplasm can be used as a new diagnosis and treatment of renal cancer gene therapy.

Objective To screen permethrin human single-chain variable region (scFv) antibody for aims of developing rapid detection kit. Methods Phage display technology was used in this study. Permethrin was solid phase coated on Nunc plate as antigen. Semi-synthetic single-chain variable region of human antibody library technology was applied, and single chain variable region was screened from phage antibody library after 3 rounds "adsorption - elution - amplification" of the selection process. 100 clones were random selected as resistance to permethrin clones , enzyme-linked immunosorbent assay (ELISA), crossreactivity and competitive inhibition experiments were used to validate permethrin binding activity with strong scFv clones from the selected phage antibody clones plasmid. The plasmid was digested with restriction enzyme Sfi Ⅰ / Not Ⅰ and subcloned into pCANTAB5E vector. After transformed into E. coli XL1BIue, the plasmid was identified by restriction enzyme analysis. Results After screening in 100 clones, 18 clones had high ELISA absorbance values ( A value) at 490nm wavelength ( A490nm), then bovine serum albumin (BSA) cross-reactions identified five weak cross-reaction. Combined with the triplicate ELISA and competitive inhibition experiment results, one positive clone was acquired at last. And this clone was subcloned into pCANTAB5E vector and transformed into competent cells XL1-Blue. Conclusion Plasmid fragment was consistent with the purpose, which provided the foundation for further study of its specific affinity.

Objective:To analyze the clinical phenotype, imaging characteristics and genetic characteristics of a family of early-onset dementia caused by a new mutation in the triggerring receptor expressing on myeloid cells 2 gene (TREM2).Methods:Clinical data were collected from a patient with early-onset dementia. Then whole exome sequencing was performed for the proband, followed by Sanger sequencing for the family members.Results:The clinical manifestations of the proband (a 49-year-old female) was personality changes, mental and behavioral abnormalities, memory loss, ataxia, and seizures. Whole-exon sequencing revealed a novel homozygous mutation in exon 2 of TREM2, namely c.154C>T (p.R52C) heterozygosity in four family members, and one patient with similar clinical manifestations was deceased. The proband′s brain magnetic resonance imaging showed bilateral frontotemporal atrophy, bilateral white matter hyperintensity, thin corpus callosum. No bone cysts of the hands and feet were found by digital radiographic imaging.Conclusions:A homozygous mutation in TREM2 gene was detected in a patient with frontotemporal dementia-like dementia, epilepsy, but without bone cysts. This mutation is probably pathogenic. This research highlights the importance of TREM2 gene mutation screening in early-onset dementia, especially in those with atypical presentations.

GJ-4 is crocin enrichments extracted from Gardenia jasminoides J. Ellis, and our previous studies have shown that GJ-4 significantly improved learning and memory impairment induced by Aβ in mice. Herein, a memory deficit model was developed by injecting okadaic acid (OA) into the lateral ventricle of mice, and the neuroprotection and underlying mechanism of GJ-4 on neuronal injury caused by Tau hyperphosphorylation were investigated. The Animal Care & Welfare Committee, Institute of Materia Medica, CAMS & PUMC has approved all procedures (No.00000318). GJ-4 at different doses was intragastric administration to mice for 16 days. Step-down test and Morris water maze test showed that GJ-4 could significantly improve OA-induced memory impairment in mice, and reduced the loss of Nissl bodies in the hippocampus of mice. GJ-4 could also decrease the phosphorylation level of Tau protein at Ser396, Thr231 and Ser404 via increasing protein phosphatase 2A (PP2A) activity and inhibiting glycogen synthase kinase-3β (GSK-3β) activity. Besides, further researches indicated that GJ-4 could inhibit the level of oxidative stress in the brain of OA mice, reduce neuronal apoptosis and inhibit the neuroinflammation mediated by activation of astrocytes in the hippocampus of mice, and eventually achieve its effects in improving learning and memory impairment in mice. According to these findings, we anticipated that GJ-4 might be a potential therapeutic drug for Alzheimer's disease.

Ten isoflavonoids were isolated from the heartwoods of Caragana changduensis Lion f. by means of various column chromatographic techniques. Based on the detailed spectral data analysis (MS and NMR), as well as comparison with the literatures, their chemical structures were determined as 7,2'-dihydroxy-8,4'-dimethoxyisoflavone (1), 4'-hydroxy-7,3'-dimethoxyisoflavone (2), 5, 7, 4'-trihydroxy-2',5'-dimethoxyisoflavone (3), prunetin (4), afrormosin (5), odoratin (6), genistein (7), texasin (8), pratensein (9), and 6,7,3'-trihydroxy-4'-methoxyisoflavone (10). Among them, compounds 1-3 and 9-10 were isolated from the Caragana genus for the first time. All the compounds were obtained from this species for the first time. In the preliminary assays, compounds 1, 2, 6, and 7 possessed significant inhibitory effects on NO production, with IC50 values of 48.12, 25.32, 62.71, 43.59 μmol x L(-1), respectively.
Animals ; Caragana ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Isoflavones ; chemistry ; isolation & purification ; pharmacology ; Macrophages ; drug effects ; metabolism ; Mice ; Molecular Structure ; Nitric Oxide ; antagonists & inhibitors ; metabolism ; RAW 264.7 Cells

Objective To investigate the diagnostic and prognostic value of serum exosomal exocirc_0023461 in coronary artery disease(CAD)related acute myocardial infarction(AMI).Methods From December 2019 to January 2022,383 patients with CAD related AMI(AMI group),200 pa-tients with chronic stable CAD but no AMI(control group),and 200 healthy individuals identified with physical examination(healthy group)were recruited in our hospital.The serum exocirc_0023461 level was determined by real-time quantitative fluorescence polymerase chain reaction.The correlation of serum exosomal circ_0023461 level with clinicopathological features and oxida-tive stress indicators was analyzed.Results The serum level of exocirc_0023461 was significantly higher in the AMI group than the control group and healthy group[3.54(1.39,9.82)vs 0.86(0.62,1.23)and 0.65(0.41,0.79),P＜0.01].ROC curve analysis showed that when the serum level of exocirc_0023461 ≥1.31,its AUC value for the diagnosis of AMI was 0.857(95％CI:0.827-0.887),with a sensitivity and specificity of 78.90％and 83.50％,respectively.Kaplan-Meier survival curve displayed that the survival time was significantly shorter in the high level AMI patients without MACE than those with low level(X2=19.390,Plog rank＜0.01).Multivariate Cox regression analysis revealed that age,peripheral artery disease and serum exocirc_0023461 were independent predictors of MACE occurrence in AMI patients during follow-up(P＜0.05,P＜0.01).Pearson correlation analysis indicated that serum exocirc_0023461 level was negatively correlated with serum GPX and SOD levels(r=-0.395,r=-0.193,P＜0.01),and positively correlated with serum MDA level(r=0.194,P＜0.01).Conclusion Serum exocirc_0023461 may be a potential biomarker for the diagnosis and prognosis of CAD-related AMI,and its mechanism seems to be associated with its regulating oxidative stress and thus affecting myocardial injury.

The purpose of the present study was to investigate the features of pathophysiological neural networks in rat temporal lobe epileptogenesis. To establish electrogenic epilepsy model, repetitive tetanization (60 Hz, 2 s, 0.4-0.6 mA) was delivered into the right dorsal hippocampus (HPC) of rat brain. Rats were divided into different groups. Experimental animals received tetanic stimulation once a day for 2, 4, 6, 8 or 10 days, respectively. Primary wet dog shakes (WEDS) of the animals were recorded daily during the stimulation to understand the development of behavioral seizures at early stage of epilepsy. The T(2)-weighted (T(2)-WI) spin-echo images were obtained from each experimental rat. The results demonstrated that T(2)-WI hyperintensity of experimental rats was observed in bilateral symmetric dorsal lateral ventricle (LV) areas at stimulating day 2 (n=4), in contralateral medial and ventral LV areas to the side of the electrode at stimulating day 6 (n=5), in contralateral ventral LV areas at stimulating day 8 (n=3), and in ipsilateral ventral LV areas at stimulating day 10 (n=4). Therefore the peak rate of primary WEDS appeared on stimulating day 4 in the experimental rats. Morphological identification demonstrated that the T(2)-WI signal abnormalities were related to the enlarged LV and pathological ventricular choroidea plexus hyperplasia. The results suggest that the development of rat brain abnormalities from dorsal LV to ventral LV at early stage of epilepsy can be measured by magnetic resonance image, which implies reorganization of pathophysiologically functional networks before kindling effect appear.
Animals ; Brain ; physiopathology ; Disease Progression ; Epilepsy, Temporal Lobe ; diagnosis ; pathology ; In Vitro Techniques ; Magnetic Resonance Imaging ; Male ; Rats ; Rats, Sprague-Dawley

The purpose of this work was to study the characteristics of rat brain abnormalities at two hemispheres at the early stage of electrogenic epilepsy. Experiments were performed on 37 male Sprague-Dawley rats. Chronically repetitive tetanization (60 Hz, 2 s, 0.4 - 0.6 mA) was used to stimulate the right dorsal hippocampus (DHPC) of the rat brain once a day for 2, 4, 6, 8 or 10 d, respectively. The T(2) weighted magnetic resonance image (T(2)-WI) were obtained from each experimental rat at the end of the experiments. Histological sections were obtained after experimentation. The results showed that the main pathologic changes at the early stage of epilepsy included: (1) T(2)-WI hyperintensification, the histological enlargement of lateral ventricle (LV) and pathological hyperplasia of ventricular choroidea plexus occurred. The pathological hyperplasia was symmetric in two hemispheres, but the LV enlargement was not. (2) Histologically enlarged LV area showed a resemblance to T(2)-WI hyperintensive area. Compared with the control rats, large T(2)-WI hyperintensive area (P=0.0259; P=0.0184; P=0.0184; P=0.0404; P=0.0259) and histologically enlarged LV area (P=0.0210; P=0.01; P=0.0100; P=0.0152) were present in chronically tetanized rats. (3) Dynamic characteristics of histologically enlarged LV area resembled to those of T(2)-WI hyperintensity area in chronically tetanized rats at different stimulating day. Lateralization of T(2)-WI hyperintensity was in accordance with that of T(2)-WI abnormal area and of histologically enlarged LV. These abnormalities were severe on the contralateral side on the stimulating day 6, or on the ipsilateral side on the stimulating day 10. These results imply characteristic propagation of brain abnormalities crossing to the opposite hemisphere at the early stage of an electrogenic rat epilepsy.
Animals ; Cerebral Cortex ; pathology ; physiopathology ; Electric Stimulation ; Epilepsy ; etiology ; pathology ; physiopathology ; Hippocampus ; physiopathology ; Magnetic Resonance Imaging ; Male ; Rats ; Rats, Sprague-Dawley ; Time Factors

OBJECTIVETo summarize the Patho-typing and the clinical manifestation of biliary cast syndrome (BCS) in patients after orthotopic liver transplantation.

METHODSThe clinical manifestation, findings,therapeutic means and efficacy of 103 patients with biliary cast syndrome after orthotopic liver transplantation were retrospectively analyzed. According to the injury level of biliary duct epithelium, patients were divided into different groups. All cases were followed up for twelve months. The place, degree and time after operation would be recorded when non-anastomotic biliary stricture was found.

RESULTSThere were 59 BCS cases in the general hospital of armed police force of China. The incidence rate of BCS was 9.1%. Many BCS patients showed symptoms such as jaundice, deep urine color, gray stools, itch of skin and fever. Some were asymptomatic. In laboratory test, the liver functional enzyme in serum were increased, the total white cell count in peripheral blood was increased either. Cholangiography via T tube of biliary tract might show filling defect. According to the change degree of the biliary tract tree, there were four types filling defect concluded from all the presentation in BCS patients. Solid obturation of biliary tract were found by the check with optical fiber choledochoscope in all BCS patients, necrosis of biliary tract epithelium were observed in partial BCS patients. According to the injury level of biliary duct epithelium (gradually aggravated), BCS patients were divided into six groups (type I, type II, type III, type IV, type V and type VI). Fourteen cases were found in type I and 18 in type II. No clinical symptom was found in these two groups, a few indicators in serum (alanine aminotransferase ALT, total bilirubin TBIL, direct bilirubin DBIL) were in normal range, and others (gamma-glutamyl transferase GGT, alkaline phosphatase ALP) were heightened in 5 patients. There was no biliary cast (BC) found anymore in the period of follow-up in two groups. No stricture was found in both group. Twenty-seven cases in type III and 23 cases in type IV, it was found there were about 33.4% patients accompanied with fever and 25.9% accompanied with jaundice in type III. Paralleled,there were about 30.4% and 34.8% patients in type IV. The liver functional enzyme in serum were found increased in both type. After supporting treatment for 3-6 months,there were 5 and 3 patients present as mild non-anastomotic biliary stricture in type III and type IV group. In the group type V, there were 13 patients. The detected liver functional indicators in serum were increased. After supporting treatment for 6-12 months,there were 4 patients present as moderate non-anastomotic biliary stricture in this group. There were 18 patients in type VI group, all indicators of the liver functional enzyme in serum before the treatment were elevated conspicuously. All patients in this group were found serious stricture up to three places that have not been sustained in the period of follow-up. Nine died of MOSF, 1 died of AOSC, 8 had undergone retransplantation. In the retransplantation patients, 4 died of post operation MOSF, 3 recovered to normal, 1 patient was found BCS once more 15 d after the retransplantation and the third-transplantation was performed 7 months after the second transplantation, no BCS was found again. The deaths total rate was 13.6%, death rate of retransplantation was 44.0%, total cure rate was 54.0%, total improvement rate was 71.0% and total stenosis rate was 29.0%.

CONCLUSIONS(1) According to the check with optical fiber choledochoscope, there are 6 types of patho-typing in BCS patients. The clinical manifestation includes jaundice and fever. The filling defect of the biliary tract tree might showed 4 appearances. (2)The patho-typing contributes to the clinical manifestation and the filling defect of the biliary tract tree.

Adult ; Aged ; Biliary Tract Diseases ; etiology ; pathology ; therapy ; Female ; Follow-Up Studies ; Humans ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Complications ; etiology ; pathology ; therapy ; Prognosis ; Retrospective Studies

Two new labdane diterpenoids were isolated from 95% ethanol extract of the leaves of Callicarpa formosana Rolfe by using silica gel column, MCI column, ODS column and HPLC. Their structures were elucidated by HR-ESI-MS, NMR and ECD spectral data. All of them are new compounds, named 13E-6β-hydroxylabda-8(17),13-dien-15-oic acid (1) and 13E-7α-hydroxylabda-8(17),13-dien-15-oic acid (2). Compounds 1 and 2 were tested for antioxidant activity, and none of them had obvious activity.