Clinicopathological analysis of benign mammary ductal cystic papillomatosis with loss of myoepithelial cells / 中华病理学杂志

Objective: To investigate the histopathological and immunohistochemical characteristics of benign apocrine cystic papillary hyperplasia of the breast with loss of myoepithelial cell layer. Methods: The clinical data, histopathological features and immunohistochemical profile of patients with benign apocrine cystic papillary hyperplasia of breast with loss of myoepithelial cell layer from January 2016 to December 2021 were examined, in which six patients were identified. Results: All six patients were female, aged 36-61 years (median 46 years), who presented with a breast mass; three cases were from the left breast and three cases were from the right breast. Microscopic examination of all cases showed breast hyperplasia with apocrine cysts, accompanied by different degrees of micropapillary and papillary hyperplasia of apocrine cells. One case was associated with lobular carcinoma in situ, and one case was associated with apocrine ductal carcinoma in situ with intraductal dissemination in adenosis. Immunohistochemical staining of CK5/6, p63, SMA, SMMHC, Calponin and CD10 showed complete absence of myoepithelial cell layer surrounding ducts in apocrine cystic papillary hyperplasia. Conclusions: The myoepithelial cells of apocrine cystic papillary hyperplasia of the breast may undergo abnormal changes and may even be completely lost. The diagnosis should be comprehensively considered along with cytomorphological and histological features to avoid overdiagnosis.

Effect of the Connexin 43 Coupling to the Biobehavior of Multiple Myeloma Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of gap junction intercellular communication (GJIC) combined by connexin43 (Cx43) and its signal to the biobehavior of multiple myeloma (MM) cells, and its possible mechanism.@*METHODS@#The mesenchymal stem cell (MSC) cells were isolated and cultured from patients with MM and normal donors. The expression of connexin43 (Cx43) in MSC cells from different sources was detected by RT-PCR and Western blot. The side population (SP) cells were sorted by flow cytometry (FCM). The effect of MSC cells from different sources to the cell cycle, Cx43 expression, colony formation in vitro, stem cell related genes expression, cytokines secretion and chemoresistance in MM SP cells as well as with or without Cx43 inhibitor 18α-glycyrrhetinic acid (18α-GA) was observed.@*RESULTS@#There was no significantly difference between the MSC isolated from normal donor and MM patients. Western blot showed that Cx43 expression in SP cells was up-regulated when the cells were incubated with MSC, and medium containing 18α-GA could partially inhibit it, moreover, it was more significant in MSC cells of MM patients. The ability of colony formation of SP cells in vitro was higher than those of MM cells and MM-MSC could promote the colony formation in a co-culture manner. The effect of MM-MSC to SP cells was down-regulated after 18α-GA was added. RT-PCR showed that there was several important stem cell-related genes including c-myc, Oct-4 Klf-4, and Sox-2 were found in RPMI 8226 cells, but those cells were up-regulated in SP cells (P0.05). Cytometry bead array assays showed that MM-MSCs could secrete high level of IL-6, but the levels of IL-6, IL-10 and TGF-β increased significantly when the MM-MSCs were co-cultured with SP cells (P<0.05), especially the levels of IL-6 and IL-10 were significantly higher than cultured alone. There was no significant change in the levels of bFGF and IL-17 before and after co-cultured. The levels of IL-6, IL-10 and TGF-β in supernatant decreased significantly after GJ inhibitor 18α-GA was added. PI/Annexin V assay showed that MM cells were sensitive to bortezomib (BTZ)-induced apoptosis, but the sensitivity for SP cells was weaker. The ratio of cell apoptosis was 75.2%±0.77% and 8.12%±0.86% (P<0.001), respectively. MM-MSC could down-regulate the cell apoptosis induced by BTZ, while the sensitivity of MM cells to BTZ could be partially recovered after GJ inhibitor was added.@*CONCLUSION@#MSC derived from MM patients can enhance GJIC to maintain its "hematopoiesis" by up-regulating the expression of Cx43 in MM cells, and at the same time promote cell proliferation and drug recistance by secreting multiple cytokines, which finally contributes to the relapse of MM.

miR-216b-5p Promotes the Migration and Invasion of LN-229 Glioma Cells by Targeting BTN3A2 / 中国生物化学与分子生物学报

Accumulating evidence indicated that microRNAs (miRNA) play an important role in tumor invasion and metastasis by regulating their target genes.However, whether the miRNA-216b-5p(miR-216b-5p ) and their target genes butyrophilin subfamily 3 member A2(BTN3A2) promote glioma invasion and metastasis is unclear.This study aims to study whether miR-216b-5p promoted migration and invasion in glioma cells by negatively regulating BTN3A2.The differential expression analysis of GSE15824 and GSE4290 was analyzed by GEO2R.We found that only BTN3A2 is up-regulated in both GSE15824 and GSE4290 (P<0.05).The gene set enrichment analysis (GSEA) analysis indicated BTN3A2 was related to many cancer-related pathways (P<0.05).The results of survival curves showed that the overall survival of patients with high expression of BTN3A2 decreased significantly (P <0.001).The expression of BTN3A2 was increased with the increase of WHO grade (P<0.05), while the expression of BTN3A2 was increased in 1p/19q uncombined deletion and IDH mutant patients (P<0.001).Western blotting results showed that BTN3A2 was up-regulated in seven glioma tissues and glioma cell lines U87, U251 and LN-229 and downregulated in the miR-216b-5p mimics group; Transwell results showed that transfection with BTN3A2 silencing plasmids(si-BTN3A2) or miR-216b-5p mimics plasmids could inhibit the ability of migration and invasion in LN-229 cells in vitro (P<0.05).The online websites predicted miR-216b-5p as a potential target gene of BTN3A2.The survival curve results show that compared with patients with low expression of miR-216b-5p , the survival rate of patients with high expression was significantly increased (P=0.025).The relative expression of miR-216b-5p was decreased in U87, U251 and LN229 cells was detected by real time quantitative PCR (P<0.05).The results of dual luciferase assay showed that BTN3A2 could bind to miR-216b-5p (P<0.05).Transwell experiment results showed that overexpression of miR-216b-5p can inhibit the migration and invasion ability of LN229 cells (P<0.05).In summary, miR-216b-5p promotes the migration and invasion by targeting BTN3A2 of LN-229 glioma cells.

Effect of M1/M2 Alveolar Macrophage Subsets in COVID-19 and Regulatory Mechanism of Traditional Chinese Medicine / 中国实验方剂学杂志

The pathological anatomical results of coronavirus disease-2019 (COVID-19) patients showed that excessive inflammatory reaction in the lungs is one of the important causes for such complications as acute lung injury or acute respiratory distress syndrome. Therefore, regulation of immune response may be an effective measure for COVID-19. Alveolar macrophages have a high heterogeneity and plasticity. The dynamic changes of subsets balance and function of M1/M2 alveolar macrophages have a significant effect on pulmonary inflammatory response during the early and late stages of infection. This paper reviews the classification and function of macrophages and explores the mechanism of alveolar macrophage in the pathological process of COVID-19 at different stages and the pharmacodynamic mechanism of traditional Chinese medicine. Besides, it provides ideas for the treatment of COVID-19 with traditional Chinese medicine and other drugs' research and development based on the regulation of macrophage polarization.

Analysis of "Gansu Prescriptions" in Prevention and Treatment of Coronavirus Disease-2019 / 中国实验方剂学杂志

Coronavirus disease-2019 (COVID-19) is an acute infectious disease caused by a 2019 novel coronavirus (2019-nCoV) infection. It is highly contagious, and can spread quickly home and abroad. It has caused a global pandemic. After the outbreak, Gansu province actively responded to the national "integrated Chinese and western medicine(ICWM)" epidemic prevention policy by organizing an expert group on the prevention and treatment of traditional Chinese medicine(TCM) and establishing a joint working mechanism of ICWM. In adherence to the principle of ICWM, it highlighted the advantages of TCM in epidemic prevention, and emphasized early, timely and whole course use of TCM. The expert group continued to summarize in practice and form a series of "Gansu prescriptions", so as to explore the prevention and control strategy of "prevention in advance, timely interruption and reversal, early prevention and cure, and cure in early stage". Before illness, the prevention shall be made in advance by taking Fuzheng Biwen prescription based on constitution differentiation, in order to strengthen the body resistance and removing pathogenic Qi, after the onset, the syndromes were first treated, interrupted and reversed, and Xuanfei Huazhuo prescription and Qingfei Tongluo prescription were administered based on syndrome differentiation, so as to exorcise pathogenic Qi and cure COVID-19 at the early stage, at the beginning stage of recovery, Jianpi Yifei prescription was used to strengthen the spleen and lungs, and harmonize the stomach and resolve dampness, so as to prevent recurrence. In the principle of ICWM, "Gansu prescriptions" were selected based on the constitution differentiation and syndrome differentiation, so as to prevent the occurrence of epidemics, block light and common symptoms from developing to heavy and critical symptoms, improve the clinical efficacy, shorten the course of disease, and reduce the incidence of critical illness, thereby reducing mortality.

Clinical Efficacy of Xuanfei Huazhuo Prescription on 40 Cases of COVID-19 / 中国实验方剂学杂志

Objective:To evaluate the clinical efficacy of Xuanfei Huazhuo prescription in the treatment of coronavirus disease-2019 (COVID-19). Method:A total of 40 patients with COVID-19 were selected and treated with Xuanfei Huazhuo prescription. The changes of body temperature, clinical symptoms, computed tomography (CT), blood routine and biochemical indexes were observed before and after treatment. Result:The 40 patients included 15 males and 25 females, with a male to female ratio of 1∶1.7. They were aged between 20-94 years old, with the average age of （43.9±16.3） years old. The course of disease was 8-23 days, with the average of （14±4.4） days. Compared with before administration, the patients' clinical symptoms, such as cough, fever, sputum, diarrhea, loss of appetite and fatigue, were all improved (P<0.05). Before treatment the traditional Chinese medicine (TCM) syndromes of patients were mainly cold dampness lung (57.5%) and cold dampness Lung (42.5%), and the tongue coating was mainly white greasy coating (52.9%). After adjuvant treatment with Xuanfei Huazhuo prescription, the fever removal time was (2.48±2.56) days; white blood cell (WBC), lymphocyte percentage (LYM%), neutrophil percentage (NEUT%), absolute value of lymphocytes (LYM #) indexes of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), total bilirubin (TBIL), ratio of glutamic oxaloacetic transaminase to glutamic pyruvic transaminase (AST/ALT) and lactate dehydrogenase (LDH) were basically restored to the normal range (P<0.05) compared with before administration. After adjuvant treatment with Xuanfei Huazhuo prescription, the results of three pharyngeal test virus nucleic acid tests were negative, and the lung CT showed that infected lesions were absorbed and all met the discharge criteria. All 40 patients met the discharge criteria and were all cured and discharged, with a cure rate of 100%. There has been no case of recurrence with a positive result of nucleic acid detection so far. The score of symptom and clinical index of patients after administration was （1.62±1.90）, which was significantly lower than that before administration (7.65±4.08, P<0.05). Conclusion:In the adjuvant treatment of COVID-19, Xuanfei Huazhuo prescription can reduce body temperature, promote the absorption of pulmonary inflammation, and improve clinical symptoms, such as fever and cough.

Antibiotic resistance spectrum and genetic characteristics of MDRSA nasal isolate among primary school students / 中国学校卫生

Objective@#To investigate the antibiotic resistance spectrum and genetic characteristics of multidrug-resistant Staphylococcus aureus(MDRSA) nasal isolate among primary school students, and to provide a scientific basis for the prevention and control of masal MDRSA resistance and the selection of clincal drugs in children.@*Methods@#Antibiotic susceptibility experiments were performed on all SA isolates of 1 705 primary school students from 8 primary schools in Guangzhou selected by using multistage cluster stratified sampling method. MDRSA antibiotic susceptibility spectrum was analyzed, and the resistant, virulence and immune evasion cluster(IEC) genes detected by polymerase chain reaction(PCR).@*Results@#The prevalence of MDRSA nasal carriage was 20.76%(354/1 705), and the proportion of multidrug resistance among SA isolates was 96.20%(354/368). The predominant resistant antibiotics of MDRSA isolates were penicillin(99.72%), erythromycin(96.33%), clindamycin(90.96%) and teicoplanin(90.11%). Notably, 240(67.80%, 240/354) MDRSA isolates were resistant to more than six antimicrobial categories. And the predominant detection rates of resistant genes were BlaZ(92.66%), Tet(M)(49.72%), virulence genes Tst(25.42%) and IEC genes Sak(92.09%), Hlb(61.58%).@*Conclusion@#We found high prevalence of nasal colonization MDRSA from healthy children. Moreover, MDRSA isolates has a high resistant rate to multiple antibiotics, and the proportion of resistant to ≥6 antimicrobial categories is high.

Protective effects of Angelica sinensis and Angelica sinensis polysaccharides against liver and kidney injury induced by irradiation in rats / 解放军医学杂志

Objective To investigate the protective effects of Angelica sinensis and Angelica sinensis polysaccharide against liver and kidney injury induced by irradiation of SD rats.Methods Thirty-two SD rats were randomly divided into four groups:control group,model group,Angelica decoction group and Angelica sinensis polysaccharide group.After 14 days of routine feeding,the rats were treated by intragastric administration once a day for 7 days.From the 8th day,the rats received whole body X-ray irradiation for 2 days.The total absorbed dose was 6Gy,and the dose rate was 92.26cGy/min.The rats were killed and the blood were collected from the femoral artery 3 days after irradiation.The content of superoxide dismutase (SOD),malondialdehyde (MDA),glutathione peroxidase (GSH-Px) and lipid peroxide (LPO) in the liver and kidney tissues was detected by colorimetric analysis.The pathological changes of the liver and kidney were observed by HE staining under microscope.The expression of Nrf2 protein in the liver and kidney of the rats was detected by Western blotting.Results Compared with control group,the weight of the liver and kidney significantly increased,the activities of SOD and GSH-Px significantly decreased,the contents of MDA and LPO significantly increased,and the liver and kidney cells showed obvious edema,and the content of Nrf2 in the liver and kidney increased in the model group.Compared with model group,the weight of the liver and kidney decreased,SOD and GSH-Px activities significantly increased,the contents of MDA and LPO significantly decreased,edema of the liver and kidney significantly reduced,the expression of Nrf2 in the liver and kidney tissues decreased in Angelica decoction group and Angelica sinensis polysaccharides group,while there was no difference between Angelica sinensis polysaccharide group and Angelica decoction group.Conclusion Angelica sinensis and Angelica polysaccharides have a good protective effect against liver and kidney injury induced by radiation in SD rats,which is implemented mainly through influencing the expression of free radicals.The protective effect of Angelica against radiation injury is achieved mainly by Angelica polysaccharide.

Protective effects of Astragalus immortal prescription against liver oxidative stress in irradiated mice / 解放军医学杂志

Objective To explore the protective effect and possible mechanism of Astragalus immortal prescription (AIP) from hepatic oxidative stress in mice irradiated by X ray.Methods Six to eight weeks old Kunming mice were randomly divided into normal control group,irradiation alone group,positive control group,and low,medium and high dose groups of AIP (10 rats in each group),physiological saline,thymus peptide and different concentration decocta of AIP were given respectively.After continuous gavage for 10 days,the whole body was irradiated with X ray at the dose of 8 Gy at a time,and then continued lavaging for 3 days,with weighing the mice and observing the diet,water intake and general condition.After killing the mice,the liver was take and the liver index was calculated;the morphological changes of the liver tissues were observed and the expressions of superoxide dismutase (SOD),malondiadehyde (MDA),glutathione peroxidase (GSH-Px) and nuclear factor erythroid 2-related factor 2 (Nrf2) in the liver tissues were detected.Results Compared with the normal control group,the weight of mice in model group decreased obviously (P＜0.01);pathological section showed that the liver cells were disordered,the central vein was congested and blocked seriously,the surrounding liver cells were denatured and necrotic;the activity of SOD and GSH-Px in liver tissues decreased,and the content of MDA increased (P＜0.05),the expression of Nrf2 protein increased significantly (P＜0.01).Compared with the radiation alone group,the weight of mice in positive control group and high dose AIP group increased significantly (P＜0.01);pathological section showed that the liver tissue structure was basically normal,the liver cells were radially arranged with the central vein as the center;the activity of SOD and GSH-Px in liver tissues increased,and the content of MDA decreased (P＜0.05),the expression of Nrf2 protein decreased significantly (P＜0.01).Conclusion AIP has protective effect from hepatic oxidative stress in mice irradiated by X ray,and the mechanism may be related to its antioxidant effects.

Invasive lobular carcinoma of basal-like subtype of breast: a clinicopathologic analysis / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathologic features, clinical progress and prognosis of the basal-like subtype of invasive lobular carcinoma (ILC) of the breast.</p><p><b>METHODS</b>Four cases of ILC were analyzed by detailed histopathologic observation and immunohistochemical staining for E-cadherin, p120 catenin, ER, PR, HER2, CK5/6, EGFR, p63, p53, Ki-67 using MaxVision method. The follow-up and clinical data were analyzed.</p><p><b>RESULTS</b>Morphologically, one case was mixed ILC and three cases were pleomorphic ILC. The tumor cells were negative for E-cadherin except one case with focal membrane positivity, and all showed p120 catenin cytoplasmic positivity except one case with focal membrane positivity. All cases were negative for ER, PR and HER2 (triple negative), and positive for EGFR and CK5/6. Two cases were positive for p63. The cases were partly and weakly positive for p53, and the Ki-67 positive rate was between 30% and 75%. Follow-up data showed that two cases developed chest wall metastases, and in one case, there was progression to liver and abdominal metastases.</p><p><b>CONCLUSIONS</b>ILC of the breast are ER, PR and HER2 "triple negative", CK5/6 and EGFR positive, indicative of basal-like characteristics. Basal-like subtype of ILC are peculiarly prone to metastasis and poor response to chemotherapy, suggesting that it is associated with poor prognosis.</p>

Tongjiang granule intervened non-erosive reflux disease patients of Gan-Wei incoordination syndrome: a curative effect observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To assess the effectiveness of Tongjiang Granule (TJG) in treating non-erosive reflux disease (NERD) of Gan-Wei incoordination syndrome (GWIS).</p><p><b>METHODS</b>Totally 128 NERD patients of GWIS were recruited from outpatients or inpatients at Department of Digestive Disease, Xiyuan Hospital, China Academy of Chinese Medical Sciences from February 2009 to November 2010. They were randomly assigned to two groups using the block group in the ratio of 1:1, 64 cases in each group. Patients in the experiment group were treated with TJG, 10 g each time, three times a day, while those in the control group were treated with Omeprazole Tablet, 20 mg each time, two times a day. The treatment course of both groups was 4 weeks. The symptoms questionnaires and SF-36 quality of life scale were observed.</p><p><b>RESULTS</b>Finally 114 patients completed the trial. There was statistical difference in epigastric upset, pharyngeal foreign body sensation, abdominal swelling or abdominal pain, and integral of excrement between the two groups before treatment (P < 0.05). However, there was no statistical difference in the rest indices between the two groups (P > 0.05). Compared with before treatment in the same group, the scores of each symptom or the total symptoms were somewhat improved in the two groups (P < 0.01, P < 0.05). There was statistical difference in the rest scores (P > 0.05) except the score of mental health in the experiment group (P < 0.01, P < 0.05). There was statistical difference in the rest scores (P > 0.05) except the score of physical function in the control group (P < 0.01, P < 0.05). There was statistical difference in post-treatment acid reflux, irritability, depression, body pain, roles of emotions (P < 0.01, P < 0.05). The total effective rate was higher in the experimental group, showing no statistical difference when compared with that of the control group (P > 0.05).</p><p><b>CONCLUSIONS</b>TJG had confirmative efficacy in treating NERD patients of GWIS. Meanwhile, it could improve their quality of life, with no obvious adverse reaction.</p>

Influence of degenerative changes of intervertebral disc on its material properties and pathology / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the material properties of normal and degenerated intervertebral discs (IVDs) and examine the effect of degenerative changes on IVD pathology.</p><p><b>METHODS</b>A computer-based online search was undertaken to identify English articles about material properties of IVDs published from January 1950 to 2011 in PubMed database. The retrieved keywords included material properties, intervertebral disc and degeneration. Based on the principles of reliability, advancement and efficiency, the obtained data were primarily examined, and the original source was retrieved to read the full-text. Repetitive articles were excluded. The data of material properties of normal and degenerated IVDs were summarized and analyzed by meta-analysis.</p><p><b>RESULTS</b>The data of Young's modulus, Poisson's ratio, shear modulus, hydraulic permeability and intradiscal pressure of normal and degenerated IVDs were obtained. Compared with normal IVDs, the Young's modulus and shear modulus of annulus fibrosus and nucleus pulposus were higher in degenerated IVDs, the Poisson's ratio was lower while the hydraulic permeability and intradiscal pressure were higher. Besides, the degeneration-related alterations in IVDs had an influence both on itself and other spinal structures, leading to diseases such as bulging disc, discogenic pain and spinal stenosis. Meanwhile, the heavy mechanical loading and injury indicated important pathways to IVD degeneration.</p><p><b>CONCLUSIONS</b>To a certain extent, the degenerative changes of IVD influence its material properties. And the degeneration-related alterations of composition can cause structural failure of IVDs, leading to injuries and diseases.</p>

Cervical spinal canal narrowing and cervical neurological injuries / 中华创伤杂志(英文版)

Cervical spinal canal narrowing can lead to injury of the spinal cord and neurological symptoms including neck pain, headache, weakness and parasthesisas. According to previous and recent clinical researches, we investigated the geometric parameters of normal cervical spinal canal including the sagittal and transverse diameters as well as Torg ratio. The mean sagittal diameter of cervical spinal canal at C(1) to C(7) ranges from 15.33 mm to 20.46 mm, the mean transverse diameter at the same levels ranges from 24.45 mm to 27.00 mm and the mean value of Torg ratio is 0.96. With respect to narrow cervical spinal canal, the following charaterstics are found: firstly, extension of the cervical spine results in statistically significant stenosis as compared with the flexed or neutral positions; secondly, females sustain cervical spinal canal narrowing more easily than males; finally, the consistent narrowest cervical canal level is at C(4) for all ethnicity, but there is a slight variation in the sagittal diameter of cervical spinal stenosis (less than or equal to 14 mm in Whites, less than or equal to 12 mm in Japanese, less than or equal to 13.7 mm in Chinese). Narrow sagittal cervical canal diameter brings about an increased risk of neurological injuries in traumatic, degenerative and inflammatory conditions and is related with extension of cervical spine, gender, as well as ethnicity. It is hoped that this review will be helpful in diagnosing spinal cord and neurological injuries with the geometric parameters of cervical spine in the future.

A randomized, controlled, double-blinded and double-dummy trial of the effect of tongjiang granule on the nonerosive reflux disease of and Gan-Wei incoordination syndrome / 中国结合医学杂志

<p><b>OBJECTIVE</b>To assess the effectiveness of tongjiang granule (TJG) on the patients with nonerosive reflux disease (NERD) of Gan-Wei incoordination syndrome, its impact on their quality of life, and its safety.</p><p><b>METHOD</b>A randomized, controlled, double-blinded, and double-dummy method was adopted in the trial. There were 120 NERD patients enrolled in the study and randomly divided into the experiment and control groups, each with 60 patients; drugs were distributed according to the drug number by patients' inclusion sequences. In the experiment group, patients were given TJG 10 g and mosapride citrate dummy 5 mg three times a day, and in the control group, patients were given mosapride citrate 5 mg and TJG dummy 10 g three times a day. The treatment courses of both groups were 4 weeks.</p><p><b>RESULTS</b>Among 120 included patients, 112 were screened for full analysis set (FAS), and 105 were screened per-protocol set (PPS). The results were as follows: (1) the improvement of total scores of symptom in the experiment group (0-4 week) were 15.93±7.88 scores by FAS and 16.22 ±7.75 scores by PPS, and they were 10.43±10.16 scores and 10.79±10.27 scores in the control group, respectively. The 95% CI of net scores improvement between the two groups were 2.10-8.90 scores and 1.92-8.94 scores in FAS and PPS; it was significantly better in the experiment group than that in the control group (P<0.05). (2) The improvement of scores of major symptom in the experiment group (0-4 week) were 10.68±5.35 by FAS and 10.89±5.29 by PPS and 7.40±7.41 and 7.60±7.46 in the control group, respectively. The 95% CI of net scores improvement in the two groups were 0.85-5.71 and 0.71-5.69 in FAS and PPS separately, and the improvement in the experiment group was significantly better than that in the control group (P<0.05). (3) The total effective rates were 86.0% and 61.8% in the experiment and the control group separately, and the Ridit analysis results showed that it was better in the experiment group (P<0.05). (4) The improvement quality of life in the domain of physical functioning and general health in the experiment group was better than that in the control group (P<0.05). (5) One case of experiment group caught a cold and recovered in six days without drug suspension. No adverse event was found in the other cases. There was no meaningful safety examination indices change in pretreatment and posttreatment periods in both groups.</p><p><b>CONCLUSION</b>TJG showed a definite effect on the treatment of NERD with Gan-Wei incoordination syndrome, and it could improve the quality of life of NERD patient without obvious toxic and side effects.</p>

Function of TTG1A in hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the biological functions of TTG1A in liver fibrosis.</p><p><b>METHODS</b>Yeast two-hybrid system was used to screen proteins associated with TTG1A. Briefly, the coding sequence of TTG1A was cloned into pGBKT7 vector, and the recombinant plasmid was transformed into yeast cells AH109 ( a type), then these cells were mated with yeast cells Y187 (a type) transformed with human leukocyte cDNA library plasmid pACT2. The obtained diploid yeast cells were plated on synthetic dropout nutrient medium containing X-alpha-gal for double selection. The plasmids from positive colonies were transformed into E.coli and sequenced.</p><p><b>RESULTS</b>The recombinant yeast expression vector pGBKT7-TTG1A was successfully constructed. Nineteen TTG1A binding proteins, including Homo sapiens major histocompatibility complex, class II DP beta 1 (HLA-DPb1), Homo sapiens ribosomal protein L30 (RPL30), Homo sapiens nucleophosmin Homo sapiens nucleobindin 2 (NUCB2), Homo sapiens ash2, variant Gaucher disease and variant metachromatic leukodystrophy, MORF4L1, Homo sapiens ubiquitin-conjugating enzyme E2L3 (UBE2L3), APOA1, Homo sapiens lectin, and galectin 1, were identified.</p><p><b>CONCLUSIONS</b>This study may help to elucidate the molecular function of TTG1A.</p>

Protein expression and function of gene 2 transregulated by hepatitis B virus pre-s1 protein and its cloning / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen proteins in leukocytes interacting with PS1TP2 by yeast-two hybrid and to view their subcellular localization in HepG2 cells.</p><p><b>METHODS</b>The function and structure of PS1TP2 were studied by bioinformatic analysis. PS1TP2 gene was amplified and cloned into plasmid pET32a (+) and pGBKT7 to construct recombinant expression vectors pET32a (+)-PS1TP2 and pGBKT7-PS1TP2. They were transduced into E. coli Rosetta strain and yeast AH109. The transformed yeast mated with yeast Y187 containing leukocyte cDNA library plasmid in a 2xYPDA medium. Diploid yeast cells were plated on a synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) for selecting twice and then screening. Then a green fluorescent protein (GFP) expression vector pEGFP-C1-PS1TP2 was established, transduced into HepG2, and its subcellular localization was studied by fluorescence microscopy and confocal microscopy.</p><p><b>RESULTS</b>Bioinformatic analysis showed that the PS1TP2 gene was located at 6q24.1, the protein was unstable and the aliphatic index was very high. After transformation of the E. coli and yeast AH109, the expression protein showed: (1) the molecular weight of the expressed product was about 41000 Da, and (2) PS1TP2 existed within the cells. Diploid yeast cells were plated on the synthetic dropout nutrient medium containing X-a-gal for selecting twice and then screening. Twenty-six colonies from blue colonies were sequenced, pEGFP-C1-PS1TP2 was successfully expressed in the HepG2 cells, and PS1TP2 was located in the cell plasma.</p><p><b>CONCLUSION</b>A prokaryotic expression vector pET32a(+)-PS1TP2 was constructed successfully and the PS1TP2 was successfully expressed in the yeast system. Genes of PS1TP2 interact with leukocyte proteins. These results bring some new clues for studying the biological functions of HBV.</p>

Screening and cloning genes transactivated by TGF beta 1 in hepatic stellate cells using suppression subtractive hybridization technique / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To construct a cDNA subtractive library of genes transactivated by TGF beta 1 in LX02 hepatic stellate cells (HSC); to screen and to clone the regulated genes transactivated by TGF beta 1; and to elucidate the molecular biological mechanism of hepatic fibrosis mediated by TGF beta 1.</p><p><b>METHODS</b>mRNA was isolated from HSC treated with TGF beta 1 or with PBS (as controls). Suppression subtractive hybridization (SSH) technique was employed to analyze the differentially expressed DNA sequence between the two groups. After restriction enzyme Rsa I digestion, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent polymerase chain reaction twice it then was subcloned into pGEM-Teasy plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5a. The cDNA was sequenced and analyzed in GenBank with Blast search.</p><p><b>RESULTS</b>The subtractive cDNA library of genes transactivated by TGF beta 1 in HSC was constructed successfully. The amplified library contained 146 positive clones, which contained 200-1000 bp of inserts. Randomly, thirty clones were analyzed by sequencing and bioinformatics, consisting of 28 known genes and 2 unknown genes.</p><p><b>CONCLUSIONS</b>The subtractive cDNA library of genes transactivated by TGF beta 1 in HSC using SSH technique was constructed successfully. Some gene coding proteins are those involved in cell growth regulation, protein synthesis, signal transduction, extracellular matrix metabolism, and anti-lipid peroxidative, which gives us some new clues for the study of the mechanism of liver fibrosis.</p>

A study on differentially expressed genes in hepatic stellate cells treated with transforming growth factor beta 1 using cDNA microarray technique / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen the differentially expressed genes in hepatic stellate cells (HSC) treated with transforming growth factor beta 1 (TGFbeta1) by cDNA microarray technique, and to elucidate the molecular pathogenesis of liver fibrosis involving TGFb1.</p><p><b>METHODS</b>Total RNA was extracted from HSC treated with TGFbeta1 and PBS by trizol and reverse-transcribed to double strand cDNA templates. Transcription of cDNA probe with biotin-labeling was performed, and then the obtained cDNA was hybridized with human cDNA microarray. The results were imaged by an Agilent scanner, and the differentially expressed genes were analyzed with bioinformatics software.</p><p><b>RESULTS</b>One hundred seventy-seven differentially expressed genes were screened from 13824 targeting genes; 123 genes were up-regulated, including connective tissue growth factor, tubulin epsilon 1, collagen, type V, alpha2, catenin delta 2, cadherin 6, type 2, Smad3, mitogen-activated protein kinase 4, growth factor receptor-bound protein 7 and MAP kinase-interacting serine/threonine kinase 1; 54 genes were down-regulated, including TNF receptor-associated factor 4, interferon regulatory factor 7, interferon inducible protein p78, bone morphogenetic protein 7, matrix gla protein, serine proteinase inhibitor, interferon stimulated gene 2.0 x 10(4), death-associated protein 6, metallothionein 1H and superoxide dismutase 2; in addition, 8 genes with unknown functions were also found.</p><p><b>CONCLUSION</b>The differentially expressed genes in HSC treated with TGFbeta1 were successfully screened by cDNA microarray technique. It revealed that the molecular pathogenesis of liver fibrosis involving TGFbeta1 was the result of co-regulation by multiple factors. This information might be of help in searching for new targets in gene therapy.</p>

Scanning electron microscopic observation: three-dimensional architecture of the collagen in hepatic fibrosis rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In the process of hepatic fibrosis, the accumulation of collagen fibers is strongly related to the hepatic function. The aim of this study was to investigate the three-dimensional architecture of the collagen network in the liver of rats with hepatic fibrosis.</p><p><b>METHODS</b>Healthy adult male Wistar rats (n = 32) were randomly divided into a control group (n = 16) and a hepatic fibrosis group (n = 16). In the control group, the rats were treated with peanut oil while the rats in hepatic fibrosis group were treated for 10 weeks with 60% CCl(4) diluted in peanut oil. The quantity of collagen fibers was detected by Western blotting; distribution of the collagen was detected by sirius red staining and polarized microscope; the three-dimensional architecture of collagen in the liver was observed under the scanning electron microscope after fixed tissues were treated with cell-maceration using NaOH. Statistical analysis was performed using the u test.</p><p><b>RESULTS</b>The quantity of collagen fibers increased significantly in the hepatic fibrosis group. With the aggravation of hepatic fibrosis, collagen fibers gradually accumulated. They interlaced the reticulation compartment and formed a round or ellipse liver tissue conglomeration like a grape framework that was disparate and wrapped up the normal liver lobule. The deposition of collagen fibers was obvious in adjacent hepatic parenchyma, especially around the portal tracts.</p><p><b>CONCLUSION</b>Our experiment showed the collagen proliferation and displays clearly the three-dimensional architecture of collagen fibers in rat liver with hepatic fibrosis by scanning electron microscope. It can provide a morphological foundation for the mechanisms of changed haemodynamics and portal hypertension in hepatic fibrosis.</p>

Investigation of RHD 1227A allele in five pedigrees in Zhejiang Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analysis the genetic mode of Rh DEL phenotype and RHD 1227A allele in Zhejiang Han population through family investigations.</p><p><b>METHODS</b>Rh DEL phenotypes were identified by a serologic adsorption-elution method. Two polymerase chain reaction-sequence specific prime (PCR-SSP) methods which detectED RHD 1227A allele and Rhesus hybrid box, respectively, and a nucleotide sequencing method focused on the exon 9 of RHD 1227A allele were employed to determine the zygosity of RHD allele.</p><p><b>RESULTS</b>All five probands with Rh DEL phenotype harbored a RHD 1227A allele and had a RHD allele deletion, and they were RHD 1227A/RHd heterozygote. One of the parent members was found to contain a RHD 1227A allele and a normal RHD allele in pedigree 1, 2 and 3, respectively. Thus, they were RHD 1227A/RHD heterozygotes and presented normal D positive phenotype. The son of proband No 1. inherited the RHD 1227A allele and presented a normal D positive phenotype due to a RHD 1227A/RHD heterozygote; The offsprings of proband No. 2, No. 4, and No. 5 did not inherit RHD 1227A allele and presented a normal D positive phenotype.</p><p><b>CONCLUSION</b>RHD 1227A allele is an important genetic marker of Rh DEL phenotype; RHD 1227A is recessive to normal RHD allele and dominant to RHd allele; RHD 1227A allele is an ancestral, but not a spontaneously mutated allele.</p>