The deubiquitinating enzyme 13 retards non-alcoholic steatohepatitis via blocking inactive rhomboid protein 2-dependent pathway

Nowadays potential preclinical drugs for the treatment of nonalcoholic steatohepatitis (NASH) have failed to achieve expected therapeutic efficacy because the pathogenic mechanisms are underestimated. Inactive rhomboid protein 2 (IRHOM2), a promising target for treatment of inflammation-related diseases, contributes to deregulated hepatocyte metabolism-associated nonalcoholic steatohepatitis (NASH) progression. However, the molecular mechanism underlying Irhom2 regulation is still not completely understood. In this work, we identify the ubiquitin-specific protease 13 (USP13) as a critical and novel endogenous blocker of IRHOM2, and we also indicate that USP13 is an IRHOM2-interacting protein that catalyzes deubiquitination of Irhom2 in hepatocytes. Hepatocyte-specific loss of the Usp13 disrupts liver metabolic homeostasis, followed by glycometabolic disorder, lipid deposition, increased inflammation, and markedly promotes NASH development. Conversely, transgenic mice with Usp13 overexpression, lentivirus (LV)- or adeno-associated virus (AAV)-driven Usp13 gene therapeutics mitigates NASH in 3 models of rodent. Mechanistically, in response to metabolic stresses, USP13 directly interacts with IRHOM2 and removes its K63-linked ubiquitination induced by ubiquitin-conjugating enzyme E2N (UBC13), a ubiquitin E2 conjugating enzyme, and thus prevents its activation of downstream cascade pathway. USP13 is a potential treatment target for NASH therapy by targeting the Irhom2 signaling pathway.

PI3K inhibitor ZSTK474 enhancing the antitumor of vesicular stomatitis virus A51 against osteosarcoma / 解剖学报

Objective To explore whether PI3K inhibitor combined with oncolytic virus can play an effective oncolytic effect on osteosarcoma. Methods The cytotoxicity to tumor cells was detected by MTT method, and the mechanism of enhancing the anti-tumor activity was explored by observation of the swelling of endoplasmic reticulum using electron microscope and the expression of apoptosis-related proteins using Western blotting. The tumor clearance ability of the combination of the PI3k inhibitor ZSTK474 and vesicular stomatitis virus A51 (VSVA51) was verified by anti-tumor experiment in vivo. The apoptosis of tumor cells was verified by immunohistochemistry. Results PI3K inhibitor could be used as sensitizers of oncolytic VSVA51, and confirmed that the)' promoted the strong apoptosis of osteosarcoma cells by aggravating the stress of endoplasmic reticulum in tumor cells (P < 0 . 01). In vivo experiments also showed that PI3K inhibitors combined with VSVA51 could significantly promote the oncolytic effect of osteosarcoma (P<0.001), and this combination therapy enhanced the infiltration of immune cells in the tumor (P<0.001). Conclusion PI3K inhibitors combined with oncolytic virus is a potential therapy for osteosarcoma.

Impact of nonsteroidal anti-inflammatory drugs on efficacy of anti-PD-1 therapy for primary liver cancer / 南方医科大学学报

OBJECTIVE@#To assess the impact of nonsteroidal anti-inflammatory drugs (NSAIDs) on clinical outcomes of patients receiving anti-PD-1 immunotherapy for hepatocellular carcinoma.@*METHODS@#We conducted a retrospective study among 215 patients with primary liver cancer receiving immunotherapy between June, 2018 and October, 2020. The patients with balanced baseline characteristics were selected based on propensity matching scores, and among them 33 patients who used NSAIDs were matched at the ratio of 1∶3 with 78 patients who did not use NSAIDs. We compared the overall survival (OS), progression-free survival (PFS), and disease control rate (DCR) between the two groups.@*RESULTS@#There was no significant difference in OS between the patients using NSAIDs (29.7%) and those who did not use NSAIDs (70.2%). Univariate and multivariate analyses did not show an a correlation of NSAIDs use with DCR (univariate analysis: OR=0.602, 95% CI: 0.299-1.213, P=0.156; multivariate analysis: OR=0.693, 95% CI: 0.330-1.458, P=0.334), PFS (univariate analysis: HR=1.230, 95% CI: 0.789-1.916, P=0.361; multivariate analysis: HR=1.151, 95% CI: 0.732-1.810, P=9.544), or OS (univariate analysis: HR=0.552, 95% CI: 0.208-1.463, P=0.232; multivariate analysis: HR=1.085, 95% CI: 0.685-1.717, P=0.729).@*CONCLUSION@#Our results show no favorable effect of NSAIDs on the efficacy of immunotherapy in patients with advanced primary liver cancer, but this finding still needs to be verified by future prospective studies of large cohorts.

Association of β-catenin expression with clinicopathological characteristics and prognosis of patients with pancreatic ductal adenocarcinoma / 肿瘤研究与临床

Objective:To investigate the expression level of β-catenin and its relationship with clinicopathology and prognosis of patients with pancreatic ductal adenocarcinoma (PDAC).Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of β-catenin mRNA in primary pancreatic cancer cell line and pancreatic ductal epithelial cell line HPDE6-C7 of the healthy. The data of 45 patients with PDAC confirmed by pathology at Xinjiang Medical University Cancer Hospital from June 2012 to December 2013 were retrospectively analyzed. Immunohistochemical method was used to detect the expression level of β-catenin in cancer tissues and adjacent tissues, and the correlation of β-catenin with pathological characteristics of patients with PDAC was analyzed. Cox proportional hazard model was performed to make univariate and multivariate analysis on the influencing factors of overall survival (OS).Results:The relative expression of β-catenin mRNA in primary pancreatic cancer cells was higher than that in HPDE6-C7 cell line [(3.83±0.83) vs. (1.00±0.03)], and the difference was statistically significant ( t = 3.45, P = 0.003). The high expression rate of β-catenin protein in PDAC tissues was higher than that in para-cancer tissues [68.9% (31/45) vs. 28.9% (14/45)], and the difference was statistically significant ( χ2 = 7.50, P = 0.005). The high expression rate of β-catenin protein in PDAC patients with different tumor diameter and TNM staging had statistically significant differences ( P = 0.026, P = 0.036). The median OS time of 45 patients was 22.5 months, and that of high expression of β-catenin protein group in 31 patients was 19 months, that of low expression of β-catenin group in 14 patients was 29 months, and the difference was statistically significant ( P = 0.009). Univariate Cox analysis showed that preoperative carbohydrate antigen199 (CA199) level, tumor diameter, tumor differentiation degree and the expression level of β-catenin protein were influencing factors of OS of patients with PDAC. Multivariate Cox analysis showed that preoperative CA199 ( OR = 9.883, 95% CI 2.815-34.689, P < 0.001), tumor diameter ( OR = 6.117, 95% CI 1.578-24.179, P = 0.009), tumor differentiation degree ( OR = 3.834, 95% CI 1.158-12.697, P = 0.028), the expression level of β-catenin protein ( OR = 0.139, 95% CI 0.045-0.430, P = 0.001) were independent affecting factors of OS of patients with PADC. Conclusions:β-catenin is abnormally highly expressed in PDAC which is correlated with the disease progression of patients and may be a new indicator and therapeutic target of prognosis for PDAC patients.

Expression of lymphoid enhancer factor-1 in pancreatic ductal adenocarcinoma and its effect on prognosis / 国际肿瘤学杂志

Objective:To investigate the expression of lymphoid enhancement factor-1 (LEF-1) in Wnt signaling pathway in pancreatic ductal adenocarcinoma (PDAC) and its significance.Methods:The relative expressions of LEF-1 mRNA in human PDAC cell line PANC-1 and normal pancreatic ductal epithelial cell line HPDE6 were detected by fluorescence quantitative PCR. A total of 45 pancreatic cancer tissue specimens and their corresponding paracancerous tissue specimens were collected from the Department of Hepatobiliary and Pancreatic Surgery, Affiliated Tumor Hospital of Xinjiang Medical University from June 2012 to December 2013. The expressions of LEF-1 in the cancer tissues and paracancerous tissues were detected by immunohistochemistry, and the relationships between LEF-1 expression and clinicopathological characteristics and prognosis of patients were analyzed.Results:Fluorescence quantitative PCR showed that the relative expression level of LEF-1 mRNA in PANC-1 cell line was significantly higher than that in HPDE-6 cell line (2.895±0.485 vs. 1.006±0.126, t=3.056, P<0.001). Immunohistochemical results showed that LEF-1 was highly expressed in 33 cases (73.3%) of cancer tissues, which was higher than that in 12 cases (26.7%) of adjacent tissues, and there was a statistically significant difference ( χ2=14.815, P<0.001). LEF-1 expression was correlated with preoperative carbohydrate antigen (CA) 19-9 level ( P<0.001) and local lymph node metastasis ( P=0.041). Survival analysis showed that the median overall survival (OS) was 22.0 months in patients with PDAC, 19.0 months in patients with high LEF-1 expression ( n=33), 31.0 months in patients with low LEF-1 expression ( n=12), and there was a statistically significant difference ( χ2=5.554, P=0.018). Univariate Cox regression analysis showed that age ( HR=1.962, 95% CI: 1.043-3.692, P=0.037), LEF-1 ( HR=2.253, 95% CI: 1.097-4.630, P=0.027), and CA19-9 ( HR=2.667, 95% CI: 1.258-5.656, P=0.011) were associated with OS. Multivariate Cox regression analysis showed that CA19-9 ( HR=6.431, 95% CI: 1.078-38.382, P=0.041), CA125 ( HR=0.151, 95% CI: 0.027-0.839, P=0.031), primary tumor size ( HR=8.364, 95% CI: 1.925-36.335, P=0.005), LEF-1 ( HR=2.281, 95% CI: 1.025-5.075, P=0.043) were independent risk factors affecting the prognosis of PDAC patients. Conclusion:LEF-1 expression is up-regulated in PDAC tissues, which is positively correlated with preoperative CA19-9 level and local lymph node metastasis, and is an independent prognostic factor in patients with PDAC.

Effect of different tooth preparation designs on the marginal and internal fit discrepancies of cobalt-chromium crowns produced by computer-aided designing and selective laser melting processes

PURPOSE@#To evaluate the impact of five different tooth preparation designs on the marginal and internal fit discrepancies of cobalt-chromium (CoCr) crowns produced by computer-aided designing (CAD) and selective laser melting (SLM) processes. @*MATERIALS AND METHODS@#Five preparation data were constructed, after which design crowns were obtained. Actual crowns were fabricated using an SLM process. After the data of actual crowns were obtained with structural light scanning, intaglio surfaces of the design crown and actual crown were virtually superimposed on the preparation. The fit-discrepancies were displayed with colors, while the root means square was calculated and analyzed with one-way analysis of variance (ANOVA), Tukey’s test or Kruskal-Wallis test (α =.05). @*RESULTS@#The marginal or internal color-coded images in the five design groups were not identical. The shoulder-lip and sharp line angle groups in the CAD or SLM process had larger marginal or internal fit discrepancies compared to other groups (P < .05). In the CAD process, the mean marginal and internal fit discrepancies were 10.0 to 24.2 µm and 29.6 to 31.4 µm, respectively. After the CAD and SLM processes, the mean marginal and internal fit discrepancies were 18.4 to 40.9 µm and 39.1 to 47.1 µm, respectively. The SLM process itself resulted in a positive increase of the marginal (6.0 – 16.7 µm) and internal (9.0 – 15.7 µm) fit discrepancies. @*CONCLUSION@#The CAD and SLM processes affected the fit of CoCr crowns and varied based on the preparation designs. Typically, the shoulder-lip and sharp line angle designs had a more significant effect on crown fit. However, the differences between the design groups were relatively small, especially when compared to fit discrepancies observed clinically.

Improved protease activity of Pixian broad bean paste with cocultivation of Aspergillus oryzae QM-6 and Aspergillus niger QH-3

BACKGROUND: The preparation of broad bean koji is a key process in the production of Pixian broad bean paste (PBP). Protease is essential for the degradation of proteins during PBP fermentation. To obtain broad bean koji with high protease activity using the cocultivated strains of Aspergillus oryzae QM-6 (A. oryzae QM-6) and Aspergillus niger QH-3 (A. niger QH-3), the optimization of acid and neutral protease activities was carried out using Box­Behnken design with response surface methodology (RSM). RESULTS: The optimum conditions were found to be as follows: inoculation proportion (X1), 3:1 (A. oryzae QM-6: A. niger QH-3, w/w); culture temperature (X2), 33°C; inoculum size (X3), 0.5% (w/w); incubation time (X4), 5 d. The acid and neutral protease activities were 605.2 ± 12.4 U/g and 1582.9 ± 23.7 U/g, respectively, which were in good agreement with the predicted values. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles revealed that the broad bean koji extracellular proteins in the case of cocultivation were richer compared to those in the case of A. oryzae QM-6 or A. niger QH-3 strain only. In addition, the free amino acids (FAAs) in the fermentation product were 55% higher in the cocultivation process than in that involving only A. oryzae QM-6, further confirming the diversity of proteases in the fermentation products. CONCLUSIONS: The optimal conditions of koji-making in PBP were obtained using RSM. The cocultivation of A. oryzae and A. niger increases the overall enzyme activities in the culture medium and the FAAs content, which would thus have potential application in the PBP industry.

Effect of Danggui Buxuetang on PKD1/NF-κB/MnSOD Signal Pathway in Bleomycin-induced Pulmonary Fibrosis in Rats / 中国实验方剂学杂志

Objective:To observe the effect of Danggui Buxuetang on lung histopathology and protein kinase D1 (PKD1), nuclear transcription factor-κB (NF-κB) and manganese superoxide dismutase (MnSOD)-mediated oxidative stress pathway in rats with pulmonary fibrosis induced by bleomycin, so as to explore the mechanism of intervention of pulmonary fibrosis.Method:Thirty-two male SPF SD rats were randomly divided into sham operation group, model group, Danggui Buxuetang group and prednisone group, with 8 rats in each group. Except the sham operation group, the other groups were prepared through the intratracheal instillation with bleomycin. After modeling for 24 h, the rats of Danggui Buxuetang group were administered with Danggui Buxuetang (0.81 g·kg-1). The rats of prednisone group were given aqueous solution of prednisone (0.005 g·kg-1). The rats of sham operation group and model group were given the same volume of saline. After 14 days of administration, blood was collected from the femoral artery, serum was separated, and the lungs were taken by thoracotomy. The pathological changes of rat lung tissues were observed by hematoxylin-eosin staining (HE) and Masson trichrome staining, and graded by Szapiel score and Ashcroft score at the same time. The content of serum malondialdehyde (MDA), and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) were determined. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure mRNA and protein expressions of PKD1, NF-κB, MnSOD.Result:Compared with the rats in sham operation group, the rats in model group had higher Szapiel scores and Ashcroft scores (P<0.05), higher serum MDA content , but lower SOD, CAT and GSH-Px activities(P<0.01), moreover, the rat lung tissues in model group had higher mRNA and protein expressions of PKD1, NF-κB and MnSOD (P<0.01) than those in sham operation group. Compared with the rats in model group, the Szapiel scores and Ashcroft scores of the rats in Danggui Buxuetang group were decreased significantly（P<0.05）. The serum MDA content was decreased significantly, and SOD, CAT, GSH-Px activities were increased, whereas mRNA and protein expressions of PKD1, NF-κB, MnSOD in the rat lung tissues were decreased（P<0.05，P<0.01）.Conclusion:Danggui Buxuetang can reduce the degree of pulmonary fibrosis by regulating the anti-oxidation pathway of PKD1/NF-κB/MnSOD mitochondrial nucleus and improving the body's antioxidant capacity.

Peroxisome proliferator activated receptor γ involved in pathogenesis of polycystic ovary syndrome / 上海交通大学学报（医学版）

Polycystic ovarian syndrome (PCOS) is one of the most common endocrine disorders in women of reproductive age. It is characterized by hyperandrogenism, rare ovulation or anovulation, and ovarian polycystic changes. Due to the heterogeneity of clinical manifestations, the pathogenesis of PCOS has not yet been fully elucidated, but genetic factors are considered to be the main pathogenesis of PCOS. Changes in epigenetic modifications such as DNA methylation and X-chromosome inactivation patterns may affect the expression of androgen receptor genes and insulin genes, thereby altering androgen activity, increasing androgen levels, and ultimately leading to PCOS. Peroxisome proliferator activated receptors (PPARs) are members of the nuclear hormone receptor superfamily. Peroxisome proliferator activated receptor γ (PPARG) is the major expression subtype of the PPARs family in ovarian granulosa cells. PPARG plays an irreplaceable physiological function in reproduction and metabolism. It is involved in ovarian steroid metabolism, ovarian tissue remodeling, granulosa cell cycle regulation and insulin-glucose metabolism. Further research on the role of PPARG in the pathogenesis of PCOS can provide a theoretical basis for the prediction, diagnosis and treatment of long-term complications, which is reviewed by this article.

Expression of Notch signaling pathway in pancreatic cancer and its role / 实用肿瘤学杂志

Objective The objective of this study was to investigate the abnormal expression of Notch signaling pathway members in pancreatic cancer and its important effect on the development of pancreatic carcinoma. Methods Affymetrix gene expres-sion microarray was used to screen the differentially expressed members of Notch signal pathway in 10 cases of pancreatic carcinoma and its adjacent tissues,and verified by real-time quantitative PCR and Western blotting. The lentivirus expression vector carrying the siRNA fragment of Jagged 2(JAG2)gene was transfected into the pancreatic cancer primary cells to construct the JAG2 gene re-pression-expressing pancreatic cancer cell line. MTT,flow cytometry and Transwell assays were used to analyze cell proliferation, changes of cell cycle and invasive transfer capabilities. Results A total of 512 differentially expressed genes were detected by Affy-metrix gene expression microarray,including 419 up-regulated genes and 93 down-regulated genes. JAG2( up-regulated expres-sion 8. 20 times),NOTCH1(up-regulated expression 3. 74 times),HES1(up-regulated expression 3. 27 times),and NOTCH2(up-regulated expression 3. 16 times)were differentially expressed in Notch signaling pathway. The results of PCR and Western blotting were consistent with those of gene chip. The growth curves of JAG2 gene repressed pancreatic cancer cells and pancreatic cancer pri-mary cells were drawn by the standard OD490 value of d1-d5 by MTT assay. JAG2 gene repressor expression vector could signifi-cantly inhibit the proliferation of pancreatic cancer cells. The cell cycle analysis showed that the apoptosis and the arrested cell cycle at the S phase were significantly increased in pancreatic cancer cells with JAG2 gene repressor expression. The invasive ability was significantly reduced in JAG2 gene repressor expression pancreatic cancer cells(P<0. 05). Conclusion Some members of the Notch signaling pathway are significantly differentially expressed in pancreatic cancer tissues,and repression of this member can affect the growth,cell cycle,invasion and metastasis of pancreatic cancer cells.

Effects of renal denervation on blood-pressure response to hemorrhagic shock in spontaneously hypertensive rats / 中华创伤杂志(英文版)

<p><b>PURPOSE</b>Renal denervation (RD) has been demonstrated to be an effective approach to reduce blood pressure for those with resistant hypertension. Yet, we aimed to explore the effect and possible mechanism of RD on blood-pressure response to hemorrhagic shock in spontaneously hypertensive rats.</p><p><b>METHODS</b>A total of 48 male spontaneously hypertensive rats were randomized to three groups: study group, sham-operation group and control group. RD was achieved by cutting off renal nerves and swabbing phenol on it. Ten weeks after RD, 8 rats in each group were sacrificed to collect the kidney and heart tissues. The remaining rats were subjected to an operation to induce hemorrhagic shock which would lead to 40% loss of total blood volume, and observed for 120 min. The serum concentration of norepinephrine was measured before and three weeks after RD.</p><p><b>RESULTS</b>The blood-pressure and norepinephrine levels were reduced significantly after RD (p < 0.05). Systolic blood pressure and diastolic blood pressure of the surgery group were higher than those in the sham and control groups at 15, 30 and 45 min after hemorrhagic shock (p < 0.05), while no significant difference was observed at 60, 90 and 120 min (p > 0.05). Additionally, the beta-1 adrenergic receptor (β1-AR) in the study group was significantly higher than those in the other two groups (p < 0.05) after hemorrhagic shock.</p><p><b>CONCLUSION</b>This study demonstrated that RD could to some extent improve blood-pressure response to hemorrhagic shock in an established model of severe hemorrhagic shock in spontaneously hypertensive rats. The mechanism might be associated with up-regulation of β1-AR.</p>

Preparation of polyclonal antibody and establishment of chemiluminescence immunoassay for aldosterone / 中国免疫学杂志

Objective:The polyclonal antibody of aldosterone (ALD) for immunoassay was developed.And a chemiluminescence immunoassay (CLIA) for the determination of ALD in human blood was established.Methods:Aldosterone oxime was prepared by chemical modification and then conjugated with BSA to prepare immunogen.Rabbit anti ALD polyclonal antibody was prepared by immunizing rabbits with the ALD-BSA.The CLIA of ALD was performed using biotin streptavidin amplification system and competition method.Results:After identification,rabbit No.3 received the highest sensitivity to ALD antibody,and the 50％ binding inhibition (IC50) value for ALD concentration was 268 pg/ml.The measuring range of CLIA method using the antibody was 62.5-2 000 pg/ml.The assay sensitivity was 23.7 pg/ml.The intra-and inter-assay coefficients of variation were 6.9％-9.5％ and 8.5％-12.7％,respectively.Analytical recovery rate was in the range of 93.1％-104.1％.The correlation coefficient between measured and expected values were 0.996 after serial dilution.Compared with radioimmunoassay kit,the correlative equation was y =0.932x+4.596,the correlation coefficient was 0.948 (n =95).Conclusion:The result of methodological identification shows that it was in line with the basic requirements of clinical application.

Effect and mechanism of ethyl pyruvate on delayed cerebral vasospasm after subarachnoid hemorrhage / 上海交通大学学报（医学版）

Objective·To observe the degree of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) at different time points, and to investigate the effects of ethyl pyruvate (EP) on CVS following SAH and its mechanism. Methods·Fifty healthy SD rats were randomly divided into five groups i.e. sham group, SAH-3 d group, SAH-5 d group, SAH-7 d group and SAH-9 d group (n=10 in each group). Sham group was built by double cisternal injection with 0.3 mL saline each time, and SAH group was built by double cisternal injection with 0.3 mL autologous blood each time. The neurological function and degree of CVS were observed. Another forty-eight healthy SD rats were randomly divided into three groups i.e. sham+saline (equal volume) group, SAH+saline (equal volume) group and SAH+EP [100 mg/(kg·d)] group (n=16 in each group). The neurological function, degree of CVS and cell apoptosis of basilar artery were observed on day 7. The expressions of p-Akt, Bax, Bcl-2 and cleaved caspase-3 were also observed on day 7. Results·CVS significantly increased on day 3, decreased on day 5, and then significantly increased to top level on day 7, and gradually decreased on day 9. Compared with sham+saline group, the neurological scores were significantly decreased in SAH+saline group on day 7. Compared with sham+saline group, CVS and cell apoptosis of basilar artery were significantly increased in SAH+saline group on day 7. Compared with sham+saline group, the expressions of Bax and cleaved caspase-3 were significantly up-regulated, while the expressions of p-Akt and Bcl-2 were significantly down-regulated in SAH+saline group on day 7. Compared with SAH+saline group, the neurological scores significantly increased in SAH+EP group on day 7. Compared with SAH+saline group, CVS and cell apoptosis of basilar artery significantly decreased in SAH+EP group on day 7. Compared with SAH+saline group, the expressions of Bax and cleaved caspase-3 were significantly down-regulated, while the expressions of p-Akt and Bcl-2 were significantly up-regulated in SAH+EP group on day 7. Conclusion·The double hemorrhage model rat has most severely CVS on day 7. EP can attenuate vasospasm after SAH, and its mechanism may be associated with inhibition of cell apoptosis. PI3K/Akt signaling pathway may be involved, which may provide a novel therapeutic target for CVS.

Development and application of a SYBR Green Ⅱ real-time RT-PCR for detection of swine hepatitis E virus / 中国人兽共患病学报

In order to establish a real-time RT-PCR based on SYBR Green Ⅱ for detection of hepatitis E virus (HEV),a pair of special primers was designed according to the conserved sequences of ORF2 in GenBank.Result showed that the standard curve of established SYBR Green Ⅱ real-time RT-PCR had a wide dynamic range from 4.10 × 102-4.10 × 108 copies/μL with a linear correlation(r2) of 0.996.The sensitivity could reach 1.00 × 102 copies/μL.The melting curve analysis using SYBR Green Ⅱ dye showed one specific peak with a melting temperature(Tm) of 84.0 C ±0.1 C.No amplification was detected from the RNA samples of porcine reproductive and respiratory syndrome virus,classial swine fever virus,transmissible gastroenteritis virus,porcine bocavirus,porcine epidemic dearrhoea virus porcine kobuvirus and porcine rotavirus by this PCR,respectively.Excellent reproducibility was obtained for detecting constructed positive plasmid DNA with intra-assay of 0.83 ％-0.94 ％ and inter-assay of 0.83％-0.94％.Further detection of 61 specimens showed that 9 of them were HEV positive,and the results of the quantitative RT-PCR were the same as that of the conventional RT-PCR.In conclusion,the real-time quantitative RT-PCR for HEV is feasible,the real-time RT-PCR established in this study will be useful for earlier rapid laboratory diagnosis and pathogenesis of HEV.

The Role of miR-106a/b Expression Levels in the Diagnosis and Prognosis of Hepatocellular Carcinoma / 中国医科大学学报

Objective To investigate the role of microRNA-106a and microRNA-106b (miR-106a/b) in diagnosis and prognosis of hepatocellular carcinoma (HCC).Methods In this study,108 HCC patients and 54 age-and sex-matched healthy controls were enrolled.Blood samples were collected from each participant,and total RNA was extracted from the plasma.We determined miR-106a/b expression levels using quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results The miR-106a/b expression levels in HCC patients were elevated compared with the healthy controls (P＜ 0.001).The ROC curve analysis showed that miR-106a/b expression levels could be used to predict the risk of HCC,with AUC values of 0.670 (95％ CI:0.573-0.768) and 0.684 (95％ CI:0.593-0.776),respectively.The miR-106a expression level in HCC patients correlated positively with hepatitis B surface antigen (HBsAg) presence (P =0.028),differentiation (P =0.025),tumor size (P =0.002),lymph node metastasis (P =0.028),and TNM stage (P =0.037).The miR-106b expression level correlated positively with HBsAg presence (P =0.003),alpha-fetoprotein (AFP) level (P =0.031),and tumor size (P =0.005).To further investigate the correlation of miR-106a/b expression levels with overall survival (OS),Kaplan-Meier curves were plotted.The results showed that HCC patients with high miR-106a expression displayed shorter OS (P =0.013).In addition,univariate and multivariate Cox proportional hazards regression showed that miR-106a was an independent risk factor for HCC prognosis.Conclusion Circulating miR-106a/b expression levels could be used as diagnostic biomarkers for HCC,and a high circulating miR-106a expression level is an independent risk factor for poor prognosis of HCC patients.

Effect of Low Molecular Polysaccharide from Agaricus Blazei on Oxidative Injury in Hippocampal Neuronal Cells of Rats / 医药导报

Objective To investigate the effect and the potential mechanisms of low molecular polysaccharide from agaricus blazei (LMPAB) on H2O2-induced oxidative injury in hippocampal neuronal cells of rats.Methods Hippocampal neuronal cells were isolated from SD rats (24 h) and grew in culture.Cultured cells were divided into normal control group (added the same amount of nutrient solution), model control group (added 500 μmol·L-1H2O2 solution) and LMPAB high, medium, low dose groups (added 20,10,5 mg·L-1 LMPAB solution, respectively, then added 500 μmol·L-1 H2O2 solution each).The hippocampal neuron cell activity was detected with MTT method.The hippocampus neuron mitochondrial membrane potential (MMP) was detected by flow cytometry.According to the reagent instruction methods, malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities were detected.Results The activities of cell, CAT, SOD, GSH-PX and MMP in normal control group and the LMPAB high dose group were significantly higher than those of model control group (P<0.01);The content of MDA in normal control group and LMPAB high dose group was significantly lower than that of model control group (P<0.01).Conclusion The protective effect of LMPAB on hippocampal neurons with H2O2-induced injury may be related with the mechanism of enhancing the neuronal antioxidative capacity.

Mechanisms for reversal of pulmonary hypertension by rapamycin in rats / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effects of rapamycin (RAP) on pulmonary hypertension (PH) in rats, and to provide new insights into medication selection for the clinical treatment of PH.</p><p><b>METHODS</b>Fifty male Sprague-Dawley rats were randomly divided into blank control, PH model, solvent control, RAP 1, and RAP 2 groups. A rat model of PH was induced by left pneumonectomy (PE) and monocrotaline (MCT). At 5 days after PH model establishment, the solvent control group and the RAP 1 group received an intramuscular injection of solvent and RAP, respectively. At 35 days after PH model establishment, the RAP 2 group received an intramuscular injection of RAP. The mean pulmonary artery pressure (mPAP) and the right ventricle/left ventricle plus septum weight ratio (RV/LV+S) were measured in each group. Histopathological changes in the right lung were evaluated by hematoxylin-eosin (HE) staining. The relative expression of alpha-smooth muscle actin (α-SMA) and smooth muscle protein 22-alpha (SM22α) in each group was determined using real-time PCR.</p><p><b>RESULTS</b>At 35 days after surgery, the PH model and the solvent control groups had significantly higher mPAP and RV/LV+S than the blank control group, while the RAP 1 and the RAP 2 groups had significantly lower mPAP than the solvent control group (P<0.05). The RV/LV+S in the RAP 1 group was significantly lower than that in the solvent control group (P<0.05); however, there was no significant difference in RV/LV+S between the RAP 2 and the solvent control groups (P>0.05). HE staining in the right lung showed the substantially thickened pulmonary artery wall and narrowed arterial lumen in the PH model and the solvent control groups compared with the blank control group. Different degrees of reversal of the pulmonary artery wall thickening were observed after RAP administration. The results of real-time PCR revealed that the relative expression of α-SMA and SM22α in the PH model and the solvent control groups was significantly lower than in the blank control group, while the relative expression of α-SMA and SM22α in the RAP 1 and the RAP 2 groups was significantly higher than in the solvent control group (P<0.05).</p><p><b>CONCLUSIONS</b>RAP can reverse the increase in pulmonary artery pressure and the right ventricular hypertrophy probably by regulation of the phenotypic conversion of vascular smooth muscle cells.</p>

Sinus histiocytosis with massive lymphadenopathy / 中国小儿急救医学

Sinus histiocytosis with massive lymphadenopathy( SHML) is also called Rosai-Dorfman disease.It is a kind of benign lymphoid tissue proliferative diseases with unknown etiology.SHML appeared mostly in children and adolescent.It has diverse clinical manifestations accompanied with multiple organ inju-ry,and no clear laboratory indicators could support the disease,being a rare disease in pediatrics,easyot miss diagnosis.Thsi article reviewde the latest progress on diagnostis and treatment of SHML,to improve teh un-derstanding of the disease.

Prognostic significance of CUEDC2 expression in hepatocellular carcino-ma / 中国病理生理杂志

AIM:To investigate the expression of CUE domain-containing 2 (CUEDC2) in hepatocellular car-cinoma ( HCC) and to analyze its clinical prognostic significance .METHODS:Total 186 formalin-fixed paraffin-embed-ded tissues obtained from surgical HCC with detailed clinicopathological and follow -up data were used .The expression of CUEDC2 was detected by immunohistochemistry .The relationships between the expression of CUEDC 2 and clinicopatholog-ical characteristics and prognosis were analyzed .RESULTS: The positive rate of CUEDC 2 in HCC was 85.5% ( 159/186), among which, the low expression was 52.2%(97/186) and the high expression was 47.8%(89/186).CUEDC2 expression was correlated with serum alpha-fetal protein (AFP) level, tumor size, tumor number, tumor differentiation and TNM stage (P<0.05).Kaplan-Meier survival curves showed that the patients with high expression of CUEDC 2 were asso-ciated with significantly shorter overall survival and recurrence-free survival than those with low CUEDC 2 expression ( P<0.05).Multivariate Cox regression analysis revealed 3 independent prognostic factors including CUEDC 2 expression, ser-um AFP and tumor number .CONCLUSION:CUEDC2 was expressed in most HCC tissues , which was relevant to tumor growth, tumor differentiation and prognosis .CUEDC2 could be a novel valuable molecular marker to predict the HCC prog-nosis.

differential expression of microRNA-143,145 in endometriosis

Background: Recent studies showed that inappropriate expression of microRNAs [miRNAs] is strongly associated with tumor-related processes in humans [2-9,11-17]

Objective: To understand the changes of miRNAs in endometriosis

Materials and Methods: With real-time RT-PCR, we investigated the miR-143 and miR-145 expression in eutopic [EU, n=2] and ectopic endometrium [EC, n=11] [from women with endometriosis] [as well as EU+EC, n=11], along with the normal endometrium [EN, n=22] [from women without endometriosis, but with leiomyoma]

Results: We did not find that the expression of miR-143 and/or miR-145 in EN or EC changed with menstrual cycle. But our results showed the miR-143 was up-regulated in EC [p=0.000] compared to EN. The miR-143 was also up-regulated in EU, but the difference did not reach statistically significance [p=0.053]. Compared to EU, the expression of miR-143 in EC was up-regulated [p=0.016]. MiR-145 had the similar characteristic to miR-143. The miR-145 was up-regulated in both EU [p=0.004] and EC [p=0.000] in compared to EN group. When compared with EU, the miR-145 in EC was up-regulated [p=0.008]

Conclusion: In conclusion, the miR-143 and miR-145 may play a certain role in the development and progression of endometriosis