Correlation between expressions of serum COL1A1/2 and intracranial aneurysm rupture / 局解手术学杂志

Objective To detect the levels of serum collagen type Ⅰ alpha 1 chain(COL1A1)and collagen type Ⅰ alpha 2 chain(COL1A2)in patients with intracranial aneurysm(IA),and explore their correlations with aneurysm rupture.Methods A total of 110 IA patients admitted to our hospital were regarded as the IA group and another 100 volunteers who underwent physical examination in our hospital were regarded as the control group.The expression levels of serum COL1A1 and COL1A2 were detected by ELISA.The IA patients were divided into the ruptured group(n=66)and unruptured group(n=44)according to the presence or absence of aneurysm rupture,and the clinical data and expression levels of serum COL1A1 and COL1A2 were compared between the two groups.The expression levels of serum COL1A1 and COL1A2 in patients with different Hunt-Hess grades were compared.The risk factors of aneurysm rupture in patients with IA were analyzed by multivariate Logistic regression analysis.The predictive value of serum COL1A1 and COL1A2 for aneurysm rupture in patients with IA were evaluated by receiver operating characteristic(ROC)curve.The correlation of serum COL1A1 and COL1A2 with Hunt-Hess grade for patients in rupture group was analyzed by Spearman correlation analysis.Results The expression levels of serum COL1A1 and COL1A2 for patients in the IA group were significantly higher than those in the control group(P<0.05).The number of patients with hypertension,diabetes mellitus,hyperlipidemia,aneurysm diameter>10 mm,and the expression levels of serum COL1A1 and COL1A2 in the rupture group were significantly more/higher than those in the unruptured group(P<0.05).The expression levels of serum COL1A1 and COL1A2 in patients with Hunt-Hess grades from Ⅲ to Ⅳ were significantly higher than those in patients with grades from Ⅰ to Ⅱ(P<0.05).The expression levels of serum COL1A1 and COL1A2 for patients in the rupture group were positively correlated with Hunt-Hess grade(r=0.562,0.414,P<0.05).Multivariate Logistic regression analysis showed that hypertension,diabetes mellitus,aneurysm diameter>10 mm,and increased expression levels of COL1A1 and COL1A2 were risk factors for aneurysm rupture in IA patients(P<0.05).The area under the curve(AUC)of aneurysm rupture predicted by serum COL1A1 and COL1A2 together was significantly higher than that predicted by COL1A1 alone(Z=1.905,P=0.028)and COL1A2 alone(Z=1.754,P=0.040).Conclusion The increased expression levels of serum COL1A1 and COL1A2 are risk factors for aneurysm rupture in patients with IA,and their combined prediction of aneurysm rupture in IA patients has certain clinical value.

Erythropoietin promotes myocardial infarction repair in mice by improving the function of Sca-1+ stem cells / 生理学报

Myocardial infarction (MI) is one of the leading causes of death in the world. With the improvement of clinical therapy, the mortality of acute MI has been significantly reduced. However, as for the long-term impact of MI on cardiac remodeling and cardiac function, there is no effective prevention and treatment measures. Erythropoietin (EPO), a glycoprotein cytokine essential to hematopoiesis, has anti-apoptotic and pro-angiogenetic effects. Studies have shown that EPO plays a protective role in cardiomyocytes in cardiovascular diseases, such as cardiac ischemia injury and heart failure. EPO has been demonstrated to protect ischemic myocardium and improve MI repair by promoting the activation of cardiac progenitor cells (CPCs). This study aimed to investigate whether EPO can promote MI repair by enhancing the activity of stem cell antigen 1 positive stem cells (Sca-1+ SCs). Darbepoetin alpha (a long-acting EPO analog, EPOanlg) was injected into the border zone of MI in adult mice. Infarct size, cardiac remodeling and performance, cardiomyocyte apoptosis and microvessel density were measured. Lin- Sca-1+ SCs were isolated from neonatal and adult mouse hearts by magnetic sorting technology, and were used to identify the colony forming ability and the effect of EPO, respectively. The results showed that, compared to MI alone, EPOanlg reduced the infarct percentage, cardiomyocyte apoptosis ratio and left ventricular (LV) chamber dilatation, improved cardiac performance, and increased the numbers of coronary microvessels in vivo. In vitro, EPO increased the proliferation, migration and clone formation of Lin- Sca-1+ SCs likely via the EPO receptor and downstream STAT-5/p38 MAPK signaling pathways. These results suggest that EPO participates in the repair process of MI by activating Sca-1+ SCs.

Qishen Yiqi Dripping Pills for Cardiovascular Diseases: Effects and Mechanisms / 中国结合医学杂志

Qishen Yiqi Dripping Pills (QSYQ) is a compound of Chinese medicine, which has been used to treat coronary heart disease and cardiac dysfunction. Its natural components include astragaloside IV, flavonoids, danshensu, protocatechualdehyde, salvianolic acid B, salvianolic acid A, ginsenosides Rg1, ginsenosides Rb1, and essential oils, etc. It exerts effects of nourishing qi and promoting blood circulation to relieve pain. In this review, the bioactive components of QSYQ and its effects for treating cardiovascular diseases and possible mechanism were summarized, providing references for further study and clinical application of QSYQ.

Advances in the efficacy and safety of scleral collagen cross-linking in the prevention and treatment of pathologic myopia / 国际眼科杂志(Guoji Yanke Zazhi)

In 2004, it was the first time that Wollensak and Spoerl had applied physical and chemical cross-linking methods to scleral tissue. They found that the biomechanical strength of cross-linked sclera, induced by riboflavin/ultraviolet A, glyceraldehyde and glutaraldehyde, could be improved and proposed that scleral collagen cross-linking is expected to be a new method for the treatment of pathologic myopia. In recent years, a series of explorations have been made on the effectiveness and adverse reactions of physical and chemical cross-linking in the prevention and treatment of pathologic myopia, including the establishment of various animal models and different myopia modeling methods, the improvement of cross-linking methods, the amelioration of the measurement of biomechanical strength of scleral tissue and the attention of biological parameters such as the thickness of retinal nerve fiber layer and the amplitude of electroretinogram in vivo. Genipin-crosslinking of the scleral collagen combined with posterior scleral contraction/reinforcement has been applied to clinical research. This review summarizes physical cross-linking and the genipin-crosslinking of scleral collagen to explore the effectiveness and safety of the methods in the prevention and treatment of the pathologic myopia.

Stromal interaction molecule 1 promotes microglia/macrophages M1 activation through endoplasmic reticulum stress after cerebral ischemia-reperfusion / 中华神经医学杂志

Objective:To investigate the influence and mechanism of stromal interaction molecule 1 (STIM1) in microglia/macrophages M1 activation after cerebral ischemia-reperfusion injury.Methods:(1) Animal experiment: 20 male C57BL/6J mice were randomly divided into sham-operated (Sham) group, middle cerebral artery occlusion and reperfusion (MCAO/R) group, MCAO/R+si-Ctrl group, and MCAO/R+si-STIM1 group ( n=5); MCAO/R models were established in mice of the latter 3 groups; empty vector control virus and STIM1 gene knockout lentivirus were transfected into mice in the MCAO/R+si-Ctrl group and MCAO/R+si-STIM1 group. The transfection efficiency of STIM1 and the expression of microglia/macrophages M1 activation marker cluster of differentiation 86 (CD86) in each group were observed. (2) Cell experiment: primary microglia were divided into Ctrl group, oxygen-glucose deprivation/re-oxygenation (OGD/R) group, OGD/R+si-Ctrl group, OGD/R+si-STIM1 group, OGD/R+solvent group, and OGD/R+4-phenylbutyric acid (4-PBA) group; OGD/R models were established in the later 5 groups; empty vector control virus and STIM1 gene knockout lentivirus were transfected into mice in the OGD/R+si-Ctrl group and OGD/R+si-STIM1 group; cells in the OGD/R+4-PBA group were pre-treated with 1 mmol/L 4-PBA for 1 h at 24 h before OGD/R modelling to inhibit endoplasmic reticulum stress (ERS), and cells in the OGD/R+solvent group were pre-treated with 0.5% dimethyl sulfoxide (DMSO) for 1 h at the same time. Reverse transcription quantitative polymerase chain reaction (RT-qPCR), ELISA, Western blotting and other methods were used to detect the levels of CD86, tumour necrosis factor-α ( TNF-α) mRNA, interleukin (IL)-1β, and ERS-related proteins (transcription factor C/EBP homologous protein [CHOP], activated transcription factor 4 [ATF4]) in these cells. Results:(1) Animal experiment: the STIM1 expression in MCAO/R+si-STIM1 group was significantly lower than that in Sham group, MACO/R group and MCAO/R+si-Ctrl group ( P<0.05); as compared with that in the MACO/R group and MCAO/R+si-Ctrl group, the number of microglia/macrophages co-expressing CD86 and Iba-1 around the ischemic foci of mice in the MCAO/R+si-STIM1 group was significantly decreased ( P<0.05). (2) Cell experiment: as compared with those in the OGD/R group and OGD/R+si-Ctrl group, the expression levels of STIM1, CD86, and TNF-α mRNA, and supernatant IL-1β content in the OGD/R+si-STIM1 group were significantly decreased ( P<0.05); as compared with those in the OGD/R group and OGD/R+si-CTRL group, the ATF4 and CHOP expression levels in OGD/R+si-STIM1 group were significantly decreased ( P<0.05); as compared with those in the OGD/R group and OGD/R+solvent group, the CD86 level, TNF-α mRNA expression level and IL-1β content in the OGD/R+4-PBA group were significantly decreased ( P<0.05). Conclusion:STIM1 affects microglia/macrophages M1 activation after ischemia-reperfusion injury by regulating ERS level.

Effect of anal draw-out laparoscopic anterior resection of rectal cancer on gastrointestinal motility and prognosis in patients with low rectal cancer / 国际外科学杂志

Objective:To investigate the effect of rectal draw-out laparoscopic anterior resection on gastrointestinal motility and prognosis in patients with low rectal cancer.Methods:A total of 140 patients with low rectal cancer who received treatment in Chongqing Ninth People′s Hospital from May 2017 to May 2018 were selected, including 82 males and 58 females, aged from 35 to 78 years with an average age of (59.33±9.12) years.According to the operation methods, all patients were divided into observation group (transanal pullout laparoscopic anterior resection of rectal cancer, n=70) and the control group (laparoscopic assisted anterior rectal resection, n=70). Independent sample t test or χ2 test were used to compare operation-related indicators, occurrence of complications, changes of fluid gastric emptying, small intestinal transport capacity, gastrin and motilin in 2 groups. Kaplan-meier survival curve was plotted to compare tumor progression-free survival (PFS) and overall survival (OS) in two groups. The two groups of PFS and OS were compared by log-rank test. Results:The operative time, intraoperative blood loss, postoperative drainage volume, and postoperative recovery time of the observation group were lower than those of the control group, the ability of liquid gastric emptying 24 h after operation, small intestine transport function at 24 h and 48 h after operation, the capacity of liquid gastric emptation, intestinal transport function 24 h and 48 h postoperatively, gastrin and motilin levels at 24 h, 48 h and 72 h postoperatively were significantly higher than those of the control group, with statistically significant differences ( P<0.05). Two years PFS (85.71% vs. 81.43%) and OS (92.86% vs. 90.00%) after surgery between the observation group and the control group were not statistically significant ( P>0.05). Conclusion:The anterior resection of rectal cancer by draw-out laparoscope is safe and radical, without increasing postoperative complications. Moreover, the recovery of gastrointestinal function is earlier than traditional laparoscopic assisted rectal cancer resection, which is conducive to improving the postoperative quality of life of patients, and is worthy of clinical promotion.

Systematic review of gut microbiota changes in patients with chronic heart failure / 中华心血管病杂志

Objective: To analyze the changes on gut microbiota and metabolic products in patients with chronic heart failure. Methods: By searching the Pubmed, EMBASE, Cochrane Library, and CNKI, Wanfang, and CMB databases from the day of built up to December 2019, we screened related literature exploring the intestinal flora of chronic heart failure patients, and systematic review was performed to study changes in intestinal flora composition, function, and metabolites among chronic heart failure patients. Results: A total of 10 articles were included to study the gut microbiota of patients with chronic heart failure in this analysis. The systematic review showed significant changes in β-diversity in patients with heart failure. The abundance of faecalibacterium, blautia, bacteroides, prevotella and anaerostipes was decreased, while the abundance of streptococcus, escherichia/shigella, veillonella, and enterobacte was increased. The increased microbial gene function in patients with heart failure included tryptophan metabolism, lipid metabolism, LPS synthesis,and so on, especially, bacterial genes related to trimethylamine oxide production increased significantly, while genes related to key enzymes producing the beneficial metabolite butyrate decreased significantly, and harmful metabolite trimethylamine oxide levels increased in chronic heart failure patients. Conclusion: There are significant changes in the structure, function and metabolites of intestinal flora in patients with chronic heart failure.

Optimal preoperative timing of indocyanine green administration for laparoscopic cholecystectomy under fluorescence navigation / 中华肝胆外科杂志

Objective:To analyze the optimal preoperative timing of indocyanine green administration to do the fluorescence imaging during laparoscopic cholecystectomy.Methods:A total of 102 patients with laparoscopic cholecystectomy from January 2019 to November 2019 were retrospectively analyzed in this study, including 42 male patients and 60 female patients with an average age of 49(15-87) years old. The preoperative timing of indocyanine green (2.5 mg/ml, 1 ml) administration was set at 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 h before surgery, 12, 7, 8, 6, 6, 7, 8, 10, 8, 8, 8, 7, 7 patients, respectively. The intraoperative fluorescence imaging and signal contrast were compared.Results:Comparing with 0.5h group, the liver fluorescence intensities in 5, 6, 7, 8, 9, 10, 11 and12 h groups were significantly decreased (all P<0.05). There were no differences in the fluorescence intensities of the gallbladder, gallbladder duct, common bile duct and common liver duct between those groups with different injection timepoints (all P>0.05), and signal contrast was significantly lower in 0.5 h group than patients in 6, 7, 8, 9, 10, 11 and 12 h groups (all P<0.05). When preoperative timing of indocyanine green administration was 7 h, the fluorescence signal contrast reached the highest values of 0.29. Conclusions:The optimal preoperative timing of indocyanine green intravenous administration for laparoscopic cholecystectomy under fluorescence navigation was 7 h at dose 2.5 mg.

Mechanism of Traditional Chinese Medicine and Its Compound on Liver Fibrosis / 中国实验方剂学杂志

Hepatic fibrosis refers to the pathological process of abnormal proliferation of intrahepatic connective tissue caused by various pathogenic factors, resulting in the excessive accumulation of extracellular matrix (ECM) in the liver and the formation of fibrous scar. Its continuous deterioration will gradually develop into liver cirrhosis, liver failure, liver cancer and other serious liver diseases. Because liver fibrosis and early liver cirrhosis can be reversed, it is very important to control the reversible process of liver fibrosis for the prevention and treatment of liver cirrhosis and liver cancer. In recent years, it has been found that traditional Chinese medicine(TCM) has the characteristics of multi-target, less toxic and side effects and good effect in the treatment of liver fibrosis. In this paper, the mechanism of anti-hepatic fibrosis of TCM and its compound was summarized. TCM can regulate transforming growth factor-β (TGF-β), platelet derived growth factor (PDGF), vascular endothelial growth factor (VEGF), connective tissue growth factor (CTGF) and other growth factors to inhibit the activation of (HSCs) and induce the apoptosis of activated HSCs, promote the expression of adiponectin and inhibit the secretion of leptin, inhibit the inflammatory reaction of liver, resist oxidant stress, inhibit the capillarization of hepatic sinusoidal endothelial cells, so as to effectively prevent the progress of liver fibrosis. Therefore, TCM can inhibit the development of liver fibrosis through multi-mechanism and multi-level, and is one of the important means to treat liver fibrosis.

Double LVIS stent intussusception assisted coils embolization in treatment of intracranial blood blister-like aneurysm / 中国介入影像与治疗学

Objective: To observe the effect of double LVIS stent intussusception assisted coils embolization in treatment of intracranial blood blister-like aneurysm (BBA). Methods: Data of 45 patients with BBA and treated by stent-assisted coils embolization were retrospectively analyzed. The patients were divided into double LVIS stent group (DLS group, 18 cases) and non-double LVIS stent group (NDLS group, 27 cases) according to the disparate therapy method. The operation outcomes, perioperative complications and follow-up results were compared between groups. Results: The immediately completely embolization rate in DLS group was 72.22% (13/18), in NDLS group was 55.56% (15/27), and the perioperative complications rate in DLS group was 16.67%(3/18), in NDLS group was 25.93%(7/27) (both P>0.05). No significant difference of the immediately completely embolization rate nor of perioperative complications rate was found between 2 groups (both P>0.05). At the 3- and 6-month follow-up, no significant difference of neurological recovery outcomes post operation was found between 2 groups (both P>0.05). The aneurysm recurrence rate in DLS group was 15.38% (2/13), lower than that in NDLS group (57.89%, 11/19) at 3-month follow-up (P=0.03). No significant difference of aneurysm recurrence rate at 6-month follow-up was found between groups (0 vs 13.33%, P>0.05). Conclusion: Double LVIS stent intussusception assisted coils embolization is safe and effective for treatment of BBA, which can significantly reduce 3-month aneurysm recurrence rate.

Application of balloon-assisted technique for intraoperative rupture during intracranial aneurysm coil embolization / 国际脑血管病杂志

Objective:To investigate the effectiveness of balloon-assisted technique for the treatment of intraprocedural aneurysmal rupture (IAR) during intracranial aneurysm coil embolization and its impact on the clinical outcomes of patients.Methods:Patients with intracranial aneurysm received coil embolization and complicated with IAR in Xijing Hospital of Air Force Medical University from January 2013 to January 2019 were enrolled retrospectively. They were divided into balloon-assisted hemostasis group and rapid packing hemostasis group according to the methods of intraoperative hemostasis. The modified Rankin Scale was used to evaluate the clinical outcomes at 3-month postoperative follow-up. A score of 0-2 was defined as a good outcome. Multivariate logistic regression analysis was used to identify the independent influencing factors of clinical outcome. Results:A total of 77 patients with IAR were enrolled, of which 46 (59.74%) used balloon-assisted hemostasis, and 31 (40.26%) used rapid packing hemostasis. In 51 patients (66.23%) with 3-month follow-up data, 32 (62.75%) had good outcomes, and 19 (37.25%) had poor outcomes. Univariate analysis showed that there were significant differences in time from IAR to treatment, time from IAR to confirmed hemostasis, postoperative Fisher grade changes, and good outcomes between the balloon-assisted hemostasis group and the rapid packing hemostasis group (all P<0.05). There were significant differences in IAR treatment methods, time from IAR to treatment, time from IAR to confirmed hemostasis, and postoperative Fisher grade changes between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that balloon-assisted hemostasis (odds ratio 0.234, 95% confidence interval 0.056-0.990; P=0.048) and time from IAR to confirmed hemostasis ≤10 min (odds ratio 0.097, 95% confidence interval 0.024-0.397; P=0.001) were the independent protective factors of the good outcomes in patients with IAR. Conclusion:Using balloon-assisted technique to treat IAR during intracranial aneurysm coil embolization can achieve satisfactory hemostatic effect and improve the clinical outcomes of patients.

Mechanism of action of intestinal flora in hepatic encephalopathy and related treatment methods / 临床肝胆病杂志

At present, hepatic encephalopathy has a relatively high mortality and thus greatly affects patients’ quality of life. This article describes the changes of intestinal flora in patients with hepatic encephalopathy and analyzes the mechanism of action of intestinal flora in hepatic encephalopathy and related treatment methods. It is pointed out that the development of hepatic encephalopathy is closely associated with intestinal flora, and clinical treatment by regulating intestinal flora has achieved a marked effect in patients with hepatic encephalopathy. In the future, the research on intestinal flora in patients with hepatic encephalopathy can be deepened to provide better regimens for the treatment of hepatic encephalopathy.

Establishment of a culture method for primary human nail matrix cells in serum-free media / 中华皮肤科杂志

Objective To establish a culture method for primary human nail matrix cells in serumfree media.Methods Nail matrix tissues were collected from 9 patients,who received nail or toe amputation and nail bed repair in Peking University Shenzhen Hospital between January 2016 and December 2016,and cultured in the serum-free DEME/F-12 media at a 37℃ incubator with an atmosphere of 5％ CO2 in air for 2-3 days.Then,primary human nail matrix cells were cultured in keratinocyte serumfree media (CnT-07),and the morphology of human nail matrix cells was observed by microscopy during the culture process.Immunofluorescence cytochemistry with anti-keratin 5 (K5) and K10 was performed to identify the acquired cells,and flow cytometry to analyze the cell purity.Results After 2 or 3 days of the culture,some cells began to crawl out from the tissue.On day 10,large cell masses were formed,some cells were morphologically similar to epithelioid cells arranged in a paving stone-like pattern,and some were flat giving a spindle-shaped or star-shaped appearance.Immunofluorescence cytochemistry showed that some cells could express both K5 and K10,which proved the existence of nail matrix cells,and 37.6％ of the cells expressed K10.Conclusion Human primary nail matrix cells could be successfully cultured by using the tissue culture method with serum-free culture media,and the nail matrix cells cultured in vitro can express both K5 and K10.

Systematic evaluation of patient safety culture assessment in nursing home / 中国实用护理杂志

Objective@#To focus on the patient′s safety culture management and related research of nursing home in China.@*Methods@#China National Knowledge Infrastructure (CNKI), China Biology Medicine disc (CBM), China Scientific Journal Database by VIP (VIP), Wanfang, PubMed, EBSCO and SpringerLink databases were searched by computer to find out all the literature about patient safety culture evaluation in nursing home. Two investigators independently screened, scrambled and cross-checked data according to inclusion and exclusion criteria.@*Results@#Finally 11 articles complied with the inclusion criteria, and conducted a descriptive study of patient safety culture assessments.@*Conclusions@#The evaluation of patient safety culture is conducive to the development of patient safety culture in nursing home. The study of patient safety culture in China′s nursing home is still in its infancy and needs to be further deepened.

Study on Curriculum System Setting of "3+2" Cut-through Nursing Major at Undergraduate Stage / 中国医学伦理学

Objective:To construct the "3+2" counterpart cut-through sectional training nursing major un-dergraduate curriculum system, which is oriented by vocational competence. Methods: The preliminary draft of"3+2" nursing undergraduate curriculum setting was established base on the literature review and expert group in-terview, and the 25 experts was conducted two rounds of expert questionnaire consultation using Delphi method. Results:Experts' opinions tended to be consistent after two rounds of consultation, the expert authority coefficient was 0 . 92 , the coordination coefficient of Kendall was 0 . 44 in the second round of expert consultation and finally established 5 curriculum groups, including total 28 courses of public elementary courses, professional basic cour-ses, professional core courses, professional oriented courses and centralized practice courses. Conclusion: It should construct the"3+2" counterpart cut-through nursing major undergraduate curriculum system, which is o-riented by vocational competence, and achieve effective connection between the knowledge structure and the quality of the nursing students, in order to provide the reference for perfecting the curriculum system of vocational educa-tion in our country.

Expression of protein kinase D1 and its phosphorylation at tyr463 and ser916 in squamous cell carcinoma, Bowen's disease and actinic keratosis / 中华皮肤科杂志

Objective To measure the expression of protein kinase D1 (PKD1),tyr463-phosphorylaed PKD1 (pPKD1-tyr463) and ser916-phos-phorylaed PKD1 (pPKD1-ser916) in squamous cell carcinoma (SCC),Bowen's disease (BD) and actinic keratosis (AK),and to explore their significance.Methods Fresh tissue samples were resected from lesions of patients with SCC (SCC group),BD (BD group) and AK (AK group),as well as from normal skin of healthy human controls (control group),and each group had a sample size of 10.Real-time RT-PCR was performed to measure the mRNA expression of protein kinase D1 gene (PRKD1),and Western blot analysis to determine the protein expression of PKD1,pPKD1-tyr463 and pPKD1-ser916.In addition,immunohistochemical study was conducted to determine the expression of PKD1,pPKD1-tyr463 and pPKD1-ser916 in another 50 paraffin-embedded skin samples of SCC,20 samples of BD,20 samples of AK and 10 normal skin samples.Results PRKD1 mRNA expression significantly differed among the control group (0.64 ± 0.09),SCC group (5.37 ± 1.06),BD group (2.69 ± 0.72) and AK group (2.43 ± 0.46) (F =21.37,P ＜ 0.05),and was significantly higher in the SCC,BD and AK groups than that in the control group (P ＜ 0.05),as well as in the SCC group than that in the AK and BD groups (both P ＜ 0.05).However,no significant difference in the PRKD1 mRNA expression was observed between the BD group and AK group (P ＞ 0.05).Immunohistochemical study showed that the total PKD1 protein and pPKD1-tyr463 in the SCC and BD groups were mainly expressed in the cytoplasm and cell membrane of spinous layer cells and atypical cells,and their expression rates were significantly higher than those in the AK group and control group (all P ＜ 0.01).The pPKD1-ser916 was only slightly expressed in some cancer nests of well-differentiated SCC tissues,but not in poorly-differentiated SCC,AK,BD tissues and normal skin tissues.In the SCC group,the expression rate of PKD1 increased with the increase of the pathological grade of SCC,and the PKD1 expression was positively correlated with pPKD1-tyr463 expression (rcc =0.479,P ＜ 0.05).Western blot results were consistent with immunohistochemical findings.Conclusion PKD1 and pPKD1-tyr463 may be involved in the development and differentiation of skin tumors derived from stratified squamous epithelium,and PKD1 may exert promotive effects on the formation of cutaneous SCC by activating the Tyr463 phosphorylation site.

Application of multiplex nested RT- PCR assay for screening the fusion genes in acute myeloid leukemia and its clinical significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical value of multiplex nested reverse transcription PCR (RT-nPCR) in screening acute myeloid leukemia(AML)fusion genes.</p><p><b>METHODS</b>A novel multiplex RT-nPCR assay was developed to detect 16 AML-related fusion genes (AML1-EVI1, AML1-ETO, AML1-MDS1, AML1-MTG16, MLL-AF9, MLL-AF6, MLL-AF10, MLL-ENL, MLL-MLL, PML-RARα, PLZFRARα, NPM1-RARα, CBFB-MYH11, DEK-CAN, SET-CAN and TLS-ERG) according to 2008 WHO classification of AML. The chromosome reciprocal translocations of 356 AML cases were detected by multiplex RT-nPCR and karyotyping. The positive samples were further confirmed by split- out PCR and FISH.</p><p><b>RESULTS</b>The fusion genes were detected in 172 patients with the positive detection rate of 48.31%(172/356), which was higher than that of karyotyping (31.46%) (χ²=70.314, P<0.01). Multiplex RT-nPCR is superior to karyotyping and FISH in identifying the rare, cryptic chromosome translocation (χ²=96.074, P<0.01).</p><p><b>CONCLUSION</b>The multiplex RT-nPCR used in this study can quickly, effectively and accurately screen the fusion genes in AML patients, which can provide important evidence for assessing diagnosis and treatment, and also provide necessary information for minimal residual disease (MRD) and prognosis.</p>

Clinical efficacy of transumbilical single-incision retrograde laparoscopic cholecystectomy / 中华消化外科杂志

Objective To investigate the clinical efficacy of transumbilical single-incision retrograde laparoscopic cholecystectomy.Me,otis The clinical data of 979 patients with gallbladder diseases who were admitted to the Shengjing Hospital of China Medical University from May 2009 to December 2012 were retrospectively analyzed.The numbers of patients who were admitted in the year of 2009,2010,2011 and 2012 were 51,265,374,289,respectively.The preoperative preparation of transumbilical single-incision retrograde laparoscopic cholecystectomy was similar to that of traditional multi-portal laparoscopic surgery.During the operation,the umbilical incision was selected.After the body and bottom of the gallbladder was dissociated,the cystic duct of gallbladder was dissociated and straightened,which was vertical to the common bile duct.After clipping the proximal part of the cystic duct of gallbladder with 2 hem-o-lock clips,the cystic duct was cut off with the ultrasonic knife,and then the gallbladder was removed.Postoperative nursing was also similar to that of traditional laparoscopic cholecystectomy.Patients were followed up via phone call or out-patient examination till March 2013.The wound infection,incisional hernia,incisional pain,cosmetic benefits were observed.Results No patient was converted to open surgery.Twenty patients were converted to multi-portal laparoscopic cholecystectomy because of severe inflammation (3 patients in 2009,5 in 2010,5 in 2011 and 7 in 2012).The mean operation time and volume of blood loss of the 959 patients were 48.5 minutes and (27 ± 25) mL.The operation time in 2009,2010,2011 and 2012 were 51.8 minutes,49.2 minutes,48.9 minutes and 46.7 minutes.The volumes of blood loss in 2009,2010,2011 and 2012 were 35.0 mL,32.1 mL,33.8 mL and 22.9 mL,respectively.The postoperative pain was slight.Forty-seven patients were administered antalgesics (5 in 2009,12 in 2010,18 in 2011 and 12 in 2012).In the 959 patients,umbilical swelling occurred in 4 patients,and was cured by disinfection treatment.Bile duct injury occurred in 3 patients from 2010 to 2011,timely repair wad done in 2 patients,and 1 was cured by drainage.The mean time of postoperative exhuast time and duration of hospital stay were 2.2 days and 4.2 days.A total of 924 patients were followed up for 1-3 months.The scar was hidden in the navel,and no incisional hernia occurred.Conclusion Transumbilical single-incision retrograde laparoscopic cholecystectomy is safe and effective with cosmetic benefits.

Effects of NANOG gene down-regulation on the apoptosis of T-cell acute lymphoblastic leukemia cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore gene expression of NANOG in T-cell acute lymphoblastic leukemia (T-ALL) cell lines and the effects of NANOG gene down-regulation on apoptosis of leukemia cells.</p><p><b>METHODS</b>Real-time PCR (RT-PCR) and Western blot were used to detect the expression level of NANOG gene and protein in MOLT-4, CCRF-HSB2 and Jurkat cells. To test the efficiency of RNA interference, MOLT-4 cells were firstly infected by lentiviral vectors, which were successfully constructed with NANOG specific shRNA. NANOG expression levels were subsequently re-evaluated by RT-PCR and Western blot. The percentages of early apoptotic cells (Annexin V⁺/7-AAD⁻) and late apoptotic cells (Annexin V⁺/7-AAD⁺) were analyzed by flow cytometry. The expression of apoptosis-related genes was also detected.</p><p><b>RESULTS</b>Both NANOG gene and protein expression was positive in MOLT-4 and CCRF-HSB2 cells. The lentiviral vectors pLB-shNANOG-1, pLB-shNANOG-2, and pLB-sh control were successfully constructed, as evidenced by the viral titers (1.83-3.12)× 10⁸ IU/ml. The experimental data on infection of MOLT-4 cells with such lentiviral vectors revealed that both shRNA interfering sequences (shNANOG-1 and shNANOG-2) could stably down-regulate NANOG gene and protein expressions. The percentages of early apoptotic cells in groups of shNANOG-1[(8.06 ± 1.61)%]and shNANOG-2[(5.67 ± 1.59)%]were significantly increased as compared to that of MOLT-4 group[(1.13 ± 0.40)%]or sh-control [(1.15±0.49)%](P<0.05). However, no statistical difference among them was observed for late apoptotic cells (P>0.05). The gene expression of TP53, PMAIP1, and CASP9 of either shNANOG-1 or shNANOG-2 group was augmented as compared to that of MOLT-4 group or sh-control (P<0.05). Reversely, a significant down-regulation of Bcl-2 gene expression was observed (P<0.05).</p><p><b>CONCLUSION</b>NANOG can be expressed in various human T-ALL cell lines. Down-regulation of NANOG can trigger leukemia cellular apoptosis through mitochondria-dependent apoptosis pathway.</p>

Construction of shRNA expression vector targeting AATF and establishment of stably transfected U937 cells / 中国实验血液学杂志

This study was aimed to construct the targeting AATF shRNA eukaryotic expression vector and establish the stably transfected U937 cell lines. The sequence of AATF mRNA was obtained from GenBank. After excluding homology, three plasmid expression vectors coding shRNA targeting 228 ∼ 249, 303 ∼ 324 and 443 ∼ 464 of AATF gene sequence were synthesized. Two terminals of shRNA carried BamHI and HindIII restriction sites. The selected nucleotides were cloned into the plasmid pSilencer 3.1-H1 neo respectively, and the resultant recombinant plasmids were named as pSA-1, pSA-2, pSA-3. The sequences of the recombinant plasmids were identified by DNA sequencing. The recombinant plasmids were transfected into the cell line U937 by electroporation with Neon(TM) Transfection System. The transfected cells were persistently screened under G418 (500 mg/L), and isolated with a limited dilution for 8 weeks. The inhibition of AATF mRNA and protein expression was respectively detected by RT-PCR and Western blot. The results indicated that RNAi eukaryotic expression vectors targeting AATF had correct reading frame and nucleotide sequence. Real-time PCR revealed that AATF shRNA effectively silenced mRNA expression of AATF. Western blot analysis found that AATF shRNA obviously suppressed protein expression of AATF (P < 0.05). It is concluded that the shRNA eukaryotic expression vector has been successfully constructed which can inhibit the expression of AATF, and the establishment of stably transfected U937 cell lines provide a original route for exploring the mechanism of AATF in human Leukemia further.