Effects of chronic restraint stress on the expression of N6-methyladenosine and related enzymes in the hippocampus of mice / 解剖学报

Objective To investigate the effect of chronic restraint stress on the expression of N6-methyladenosine (m6A)and related enzymes in the hippocampus of mice. Methods Twenty C57BL/6J male mice were randomly divided into control group and chronic restraint stress (CRS) group, the model group was given for 3 weeks chronic restraint stress to establish a mouse anxiety model. Open field test and elevated plus maze test were used to detect anxiety-like behavior; Immunohistochemistry and m6A RNA methylation assay were used to detect the expression changes of mouse hippocampal m6A; Western blotting and Real-time PCR were used to analyze hippocampal m6A related enzymes expression. Results 1.The behavioral results showed that, compared with the control group, the CRS group showed significantly reduced time spent in the center of the open field(P<0.01), the CRS group showed significantly reduced exploration time in the open arm of elevated plus maze (P<0.0001); 2. Immunohistochemical results showed that, compared with the control group, the hippocampal m6A content in the CRS group reduced significantly (P < 0.001); The results of the m6A RNA methylation assay showed that, compared with the control group, the CRS group showed significantly reduced amount of hippocampal m6A(P<0.05); 3. Real-time PCR results showed that the expression of hippocampal demethylase anaplastic lymphoma kinase B(AlkB) homolog 5(ALKBH5) (P<0.001) and fat mass and obestity associated protein(FTO) (P< 0.05) in the CRS group significantly up-regulated, the expression of methylase Wilms' tumour 1-associating protein (WTAP) (P<0.05) was significantly down-regulated compared with the control group; The expression of m6A methylation binding protein YTH domaincontaining family protein 3 (YTHDF3) (P < 0.05) and YTH domaincontaining protein 2 (YTHDC2) (P < 0.01) was significantly up-regulated. Western blotting result showed that, compared with the control group, the mouse hippocampal demethylase ALKBH5 (P < 0.05) and FTO (P < 0.05) expression in the CRS group significantly up-regulated, the expression of WTAP (P<0.01) was significantly down-regulated; m6A methylation binding protein YTHDF3 (P<0.01) and YTHDC2 (P<0.05) were significantly up-regulated. Conclusion In the anxiety model induced by chronic restraint stress, the expression of m6A in the hippocampus of mice is down-regulated. The mechanism may be related to the up-regulation of the m6A demethylase ALKBH5 and FTO or the down-regulation of the methylase WTAP.

Effect of Buyang Huanwu Decoction on cerebral ischemia-reperfusion injury in rats by regulating autophagy through PI3K/AKT pathway / 中国药理学通报

Aim To explore the effect of Buyang Huanwu Decoction on cerebral ischemia-reperfusion injury in rats by regulating autophagy through PI3K/AKT pathway. Methods The rats were randomly divided into five groups(n=10): sham operation group(Sham), model group(Model), Buyang Huanwu Decoction group(BYHWD), PI3K inhibitor group(LY294002)and Vehicle group(Vehicle). Except Sham group, the other groups were treated with 2h ischemia and 72 h reperfusion for modeling. The Zea Longa score was used to assess the neurological defects, HE was used to observe brain injury in the ischemic penumbra(IP), immunofluorescence was employed to detect LC3, and Western blot was used to detect pathway and autophagy marker proteins. Results Compared BYHWD group with model group, the neurological score of rats decreased, cerebral infarction volume decreased, the pathological lesions of brain IP were relieved, PI3K and p-AKT/AKT expression increased, and LC3Ⅱ/ decreased and p62 increased(P<0.05). The regulatory effect of BYHWD was weakened by LY294002(P<0.05). Conclusion Buyang Huanwu Decoction alleviates cerebral ischemia-reperfusion injury in rats by activating PI3K/AKT pathway to inhibit autophagy.

Cytopathological characterization of ascites for the diagnosis of serous ovarian carcinoma / 中华肿瘤杂志

Objective: To investigate the cytomorphological and immunocytochemical features of tumor cells in the ascites of ovarian plasmacytoma (SOC). Methods: Specimens of serous cavity effusions were collected from 61 tumor patients admitted to the Affiliated Wuxi People's Hospital of Nanjing Medical University from January 2015 to July 2021, including ascites from 32 SOC, 10 gastrointestinal adenocarcinomas, 5 pancreatic ductal adenocarcinomas, 6 lung adenocarcinomas, 4 benign mesothelial hyperplasia and 1 malignant mesothelioma patients, pleural effusions from 2 malignant mesothelioma patients and pericardial effusion from 1 malignant mesothelioma. Serous cavity effusion samples of all patients were collected, conventional smears were made through centrifugation, and cell paraffin blocks were made through centrifugation of remaining effusion samples. Conventional HE staining and immunocytochemical staining were applied to observe and summarize cytomorphological characteristics and immunocytochemical characteristics. The levels of serum tumor markers carbohydrate antigen 125 (CA125), carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) were detected. Results: Of the 32 SOC patients, 5 had low-grade serous ovarian carcinoma (LGSOC) and 27 had high-grade serous ovarian carcinoma (HGSOC). 29 (90.6%) SOC patients had elevated serum CA125, but the difference was not statistically significant between them and patients with non-ovarian primary lesions included in the study (P>0.05); The serum CEA was positive in 9 patients with gastrointestinal adenocarcinoma and 5 patients with lung adenocarcinoma, and the positive rate was higher than that in SOC patients (P<0.001); The serum CA19-9 was positive in 5 patients with gastrointestinal adenocarcinoma and 5 patients with pancreatic ductal adenocarcinoma, and the positive rate was higher than that in SOC patients (P<0.05). The serum CA125, CEA and CA19-9 were within the normal range in 4 patients with benign mesothelial hyperplasia. LGSOC tumor cells were less heterogeneous and aggregated into small clusters or papillary pattern, and psammoma bodies could be observed in some LGSOC cases. The background cells were fewer and lymphocytes were predominant; the papillary structure was more obvious after making cell wax blocks. HGSOC tumor cells were highly heterogeneous, with significantly enlarged nuclei and varying sizes, which could be more than 3-fold different, and nucleoli and nuclear schizophrenia could be observed in some cases; tumor cells were mostly clustered into nested clusters, papillae and prune shapes; there were more background cells, mainly histiocytes. Immunocytochemical staining showed that AE1/AE3, CK7, PAX-8, CA125, and WT1 were diffusely positively expressed in 32 SOC cases. P53 was focally positive in all 5 LGSOCs, diffusely positive in 23 HGSOCs, and negative in the other 4 HGSOCs. Most of adenocarcinomas of the gastrointestinal tract and lung had a history of surgery, and tumor cells of pancreatic ductal adenocarcinoma tend to form small cell nests. Immunocytochemistry can assist in the differential diagnosis of mesothelial-derived lesions with characteristic "open window" phenomenon. Conclusion: Combining the clinical manifestations of the patient, the morphological characteristics of the cells in the smear and cell block of the ascites can provide important clues for the diagnosis of SOC, and the immunocytochemical tests can further improve the accuracy of the diagnosis.

Blood glucose related adverse drug reaction of antitumor monoclonal antibodies: a retrospective analysis using Vigibase

Abstract On the increasing prevalence of using mAbs (monoclonal antibodies) in cancer therapy and the severe risk of hyperglycemia, we aimed to analyze the main clinical ADRs of mAbs, with a focus on adverse hyperglycemic events associated with currently clinically used mAbs. mAbs as well as target information were selected from Martinadale book and published articles. Drug approving information was collected from each government website, and ADR statistic data were collected from VigibaseR, comparing with Adverse Event Reporting System of US FDA. Top 10 mAbs were classified within listing in total ADR records, ADRs per year, hyperglycemic ADR records. Vigibase data were updated onto 15 Feb 2019. 20 mAbs were analyzed with 263217 ADR reports, wherein 16751 records on Metabolism and nutrition disorders and 1444 records on Glucose metabolism disorders. The geographic, age, gender distributions and annual ADR report numbers were listed respectively. Of the top 10, Rituximab, Bevacizumab and Nivolumab were on the top 3 in total ADR record and hyperglycemic record. Top 3 record results were similar in Vigibase and FDA database. It is of increasing importance for clinicians to be aware of early detection, patient management, or drug selection strategies when using mAbs, particularly within the high glycemic risk-reported mAbs, to improve the efficacy and tolerability of mAbs regiment and optimize patient outcomes.

Investigation of the API distribution homogeneity in lyophilized product-pemetrexed disodium for injection by confocal micro-Raman spectroscopy mapping / 药学学报

The method of homogeneity evaluation for active pharmaceutical ingredient (API) spatial distribution in lyophilized product was investigated for the first time with confocal micro-Raman spectroscopy mapping, using pemetrexed disodium for injection as a model drug. Certain areas of the lyophilized product were scanned to obtain Raman spectra. The classical method ("peak clipping" method) was employed for mapping with characteristic Raman peaks of the API and the excipient. Due to the API being finely dispersed in the excipient in lyophilized products, the classical method cannot discriminate between the two ingredients making the distribution homogeneity difficult to evaluate. The "ratio of characteristic peak intensities" method was then utilized. Using this method, the relative intensity of the characteristic Raman peaks of the API to the excipient was applied for mapping and the relative content of API to excipient was calculated for a homogeneity evaluation of the drug distribution. The validation of this method showed a good linear relationship between the relative intensity and the relative content of API to excipient (r2 > 0.99), and the precision and recovery were adequate for homogeneity evaluation of API by Raman spectroscopy mapping. Five products of pemetrexed disodium for injection from different manufacturers were tested through Raman maps applying this method and the histograms of relative Raman intensity were also plotted by frequency to help the homogeneity evaluation of drug distribution. The results showed that there were obvious differences in the drug distribution homogeneity from different products, where a more homogeneous API distribution was found in the brand product. This research provides a reliable method for the homogeneity evaluation of API distribution, which facilitates quality evaluation and process optimization of lyophilized products.

Clinical effect of plum-blossom needle tapping at three meridians of wrist on wrist joint contracture after stroke / 中国针灸

OBJECTIVE@#To compare the therapeutic effect of plum-blossom needle tapping at three meridians of wrist combined with rehabilitation training and simple rehabilitation training on wrist joint contracture after stroke.@*METHODS@#A total of 72 patients with wrist joint contracture after stroke were randomized into an observation group and a control group, 36 cases in each one. In the control group, simple rehabilitation training was applied, 5 times a week, 3 weeks as one course and totally 3 courses were required. On the basis of the treatment in the control group, plum-blossom needle tapping at three meridians of wrist was adopted in the observation group. The tapping regions were wrist traveling parts of three meridians of hand, ranging from up 3 to below 1 of wrist crease, 3 times a week, 3 weeks as one course and totally 3 courses were required. The active range of motion (AROM) of active wrist extension, Fugl-Meyer score (FMA) and Barthel index (BI) score were observed before and after treatment in the two groups.@*RESULTS@#The AROM, FMA scores and BI scores after treatment in the two groups were superior to before treatment (<0.05), and the improvements of 3 indexes in the observation group were superior to the control group (<0.05).@*CONCLUSION@#The therapeutic effect of plum-blossom needle tapping at three meridians of wrist combined with rehabilitation training is superior to simple rehabilitation training on wrist joint contracture after stroke.

Application of immunoassay techniques in glyphosate residue detection / 中国免疫学杂志

Glyphosate is widely used in agricultural production as an efficient herbicide.At the same time,the residual problems of glyphosate and its potential harm are gradually attracting people′s close attention.In recent years,glyphosate residue detection method has become a research hotspot,fluorescence immunoassay,enzyme-linked immunosorbent assay and immunosensor are developed rapidly by sensitive,specific and fast characteristics.This paper reviews the immunoassay techniques of glyphosate residue and provides a reference for the rapid detection of glyphosate residue.

Influence of acute high altitude exposure and short-term acclimation on platelet-associated parameters in healthy young man / 解放军医学杂志

Objective To explore the effect of high altitude exposure and short-term acclimation on the platelet-associated parameters by studying the changes of platelet-associated parameters in healthy young man.Methods Four hundred and sixtytwo young men were recruited from Chengdu (the elevation of 500m) during Jun.2012 to Aug.2013 according to the inclusion and exclusion criteria,of which 193 had been living in Chengdu (plain group),and 269 flew from Chengdu to Lhasa (3700m) in 2 hours,and then 147 of them were exposed in the high altitude for 1 day (acute high altitude exposure group),and another 122 for 7 days (short-term acclimation group).The demographic data were collected and the blood routine and platelet-associated parameters were measured of all the participants,and then the information collected were compared between the 3 groups.Results Compared to the plain group,the platelet count (PLT),platelet distribution width (PDW) and plateletcrit (PCT) reduced and the mean platelet volume (MPV) increased significantly (P＜0.05) in the acute high altitude exposure group;while all the indexes in short-term acclimation group returned to approach the levels in the plain group,but statistical differences still existed in PLT,PDW and PCT (P＜0.05).The platelet activating factor (PAF) and epinephrine (Epi) decreased markedly in acute high altitude exposure group than in plain group (P＜0.05);while in short-term acclimation group,the PAF returned to approach the level in plain group,and the Epi was further down (P＜0.05).In addition,no marked difference of 5-HT level was observed in the 3 groups (P＞0.05).Pearson correlation analysis indicated that high altitude-induced reduction of oxygen saturation (SpO2) was positively related to the changes of PLT.Conclusions Acute high altitude exposure may reduce PLT,PDW and PCT levels,but elevate MPV and enhance the platelet activity.The reduction of SpO2 might be associated with the changes of PLT,PDW and PCT.Platelet-associated parameters may be recovered to normal with a compensatory effect after short-term acclimation.

Emerging Roles of Histidine Triad Nucleotide Binding Protein 1 in Neuropsychiatric Diseases / 中国医学科学院学报

The histidine triad nucleotide binding protein1(HINT1),which belongs to the histidine triad(HIT) enzyme superfamily,exerts its enzymic activities as hydrolase or transferase. Its physiological functions are still unclear. HINT1 protein is expressed in various tissues and plays an important role in transcription and signal transduction. Earlier studies have identified HINT1 as a haplo-insufficient tumor suppressor. Other evidences indicate that HINT1 is involved in a wide variety of physiological processes,some of which are irrelevant with its basic enzymic activities. Investigations recently suggest that HINT1 is closely related to many peripheral and central nervous system diseases,and plays a vital role in some of neuropsychiatric diseases such as inherited peripheral neuropathies,schizophrenia,mood disorder,drug addiction,and Down's syndrome. In this review,the role of HINT1 in above-mentioned neuropsychiatric disorders was summarised,and the research findings of HINT1 in each of the above diseases were summarized and analyzed,in order to provide some guidance for further research on this protein.

Histidine triad nucleotide-binding protein 1 and human diseases / 中国医学科学院学报

Histidine triad nucleotide-binding protein 1 (HINT1) is a member of a superfamily of histidine triad proteins named by the conserved nucleotide-binding motif histidine-x-histidine-x-histidine-xx, in which x represents hydrophobic amino acid. HINT1 is implicated in pathological progress of many human diseases including cancer and schizophrenia; however, little is known about the essential role and pathological consequences of HINT1 in cellular physiology and diseases. Therefore, we summarize the structure, distribution, and physiological function of HINT1 in cells and tissues as well as the correlation between HINT1 and human diseases.

Down-regulation of lung resistance related protein by RNA interference targeting survivin induces the reversal of chemoresistances in hepatocellular carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Both survivin and lung resistance related protein (LRP) are related to the chemoresistances in hepatocellular carcinoma (HCC). But the relationship between survivin and LRP is indefinite. The aim of this study was to investigate the effects of down-regulation of survivin on LRP expressions and the reversal of chemoresistances in HCC both in vitro and in vivo.</p><p><b>METHODS</b>The expressions of survivin were detected by RT-PCR and Western blotting in HCC cell line SMMC-7721 and SMMC-7721/ADM. The sensitivities of these two cell lines to ADM were evaluated by MTT assays. SiRNA which targeted survivin was transfected into SMMC-7721/ADM cells, then the sensitivity of SMMC-7721/ADM cells to ADM and the expressions of survivin and LRP were detected respectively. SMMC-7721/ADM cells were transplanted subcutaneously into nude mice to establish xenograft tumors. Antitumor activities of RNA interference (RNAi) targeting survivin, various doses of ADM and combination therapies were observed respectively. Possible toxicities were evaluated. LRP expression changes were tested. Student's t test was used for evaluating statistical significance.</p><p><b>RESULTS</b>The expressions of survivin in SMMC-7721/ADM cell line showed significant elevation compared to those in SMMC-7721 cell line (P < 0.05). Positive siRNA down-regulated the expressions of survivin significantly (P < 0.05). SiRNA targeting survivin could sensitize SMMC-7721/ADM cells to ADM and down-regulate the expressions of LRP significantly (P < 0.05). Growths of the tumors were significantly inhibited in positive siRNA group as compared with those in the control group from the 8th day (P < 0.05). Combination therapies caused significant tumor inhibitions compared with tumors of nude mice in the other three groups respectively (P < 0.05). No toxicities were found in nude mice treated by siRNA and combination therapies. The expressions of LRP were markedly reduced in tumors treated with siRNA targeting survivin (P < 0.05).</p><p><b>CONCLUSIONS</b>Down regulation of survivin gene by RNAi can increase chemosensitivity of HCC both in vitro and in vivo. The reversal of drug resistance may be reduced through the inhibitions of LRP.</p>

Pathologic and immunohistochemical study on lethal primary brain stem injury / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the histopathologic changes of primary brain stem injury and to investigate their significance in the diagnosis of primary brain stem injury.</p><p><b>METHODS</b>Sixty-five autopsy cases died of primary brain stem injury and other diseases were enrolled into this study. The cases were subdivided into brain stem injury group (n = 25) and control group (including 20 cases died of cardiovascular disease and 20 cases died of non-cardiovascular diseases). The brain stem tissue sections were stained with hematoxylin-eosin and silver impregnation techniques. Immunohisto chemical study for glial fibrillary acidic protein, neurofilament, amyloid-beta and myelin basic protein was carried out. The widest cross diameters of 10 axons highlighted by immunostaining were measured in each low power field (x 100) through light miscroscopy in all the cases studied.</p><p><b>RESULTS</b>In comparing with that of the control group, there were differences in the degree of contusion lesion, reactive astrocytosis, edema and pathologic changes of neuronal cells present in the brain stem injury group and was statistically significant (P < 0.05). The axons locating in the brain stem injury group showed a distinctive histology by the appearance of significantly larger diameters (P < 0.05).</p><p><b>CONCLUSIONS</b>Primary brain stem injury demonstrates certain distinctive histopathologic changes and measurement of axonal diameters provides an additional quantitative index useful in autopsy diagnosis.</p>

Expression of P504S,CK34?E12,p63 and PSA immunohistochemistry in pathological diagnosis of prostatic adenoearcinoma / 中华泌尿外科杂志

Objective To investigate the utility of P504S(?-methylacyl-COA racemase), CK34?E12,p63 and PSA immunohistochemistry in the pathological diagnosis of prostatic adenocarcinoma (PAC).Methods The specimens of 46 cases of PAC,8 cases of prostatic high-grade intraepithelial neo- plasia (HGP1N) and 35 cases of benign prostatic hyperplasia (BPH) tissues were immunohistochemically stained with P504S,CK3413E12,p63 and PSA antibody,respectively.Results Of the 46 PAC cases,42 (91.3%) cases showed positive for P504S,including 25 cases (54.3%) who showed strongly and diffusely positive (+++) for cytoplasmic staining.In 7 (87.5%) of the 8 HGPIN cases,the specimens were also positive for P504S,and only 1(2.9%)of the 35 BPH cases showed focally weakly positive (+) for P504S.All the 8 HGPIN cases (100%) and 33 (94.3%) of the 35 BPH cases showed positive for CK34?E12 and p63,while all the 46 PAC cases showed negative for CK34?E12 and p63.In 44 (95.7%) of the 46 PAC cases,the specimens were positive staining for PSA.Conclusions P504S has high sensitivity and good specificity in the diagnosis of PAC.P504S staining in combination with HE,CK34?E12,p63 and PSA staining can improve the accurate diagnosis of PAC.