The two hybridoma cell lines secreting antikeratin monoclonal antibodies (McAb)were obtained from fusion between NS_1 myelema cells and spleen cells of BALB/cmice immunized with keratin prepared from human callus.The two cell lines(AF_2and AF_5)have a chromosome number of 97 and 96 respectively.By immunodiffusion,both AF_2 and AF_5 McAb belong to mouse IgG_1 subclass.They have been successivelypassaged for over six months in vitro,and can grow well in the peritoneal fluid ofBALB/c mice,both produce high titer of specific antibodies. To determine their specificity with keratin of epithelia,AF_2 and AF_2 McAbwere used to stain various human normal and neoplastic tissue sections by the ABC(Avidin-Biotin peroxidase complex)immunohistochemical techniques.The results showthat AF_2 and AF_5 McAb can stain a series of normal epithelial tissues(in epidermis,positive for cells above the basal layer)and epithelial tumor tissues.No staining wereobserved in nonepithelial normal and tumor tissues.These results suggest that the twoMcAb are quite specific for a definite number of cytokeratins and promise to be use-ful in the differential diagnosis of tumors.

Objective:To observe the clinical effect of marzulene-S granules combined with phloroglucinol injections in the treat-ment of pregnant women with acute gastritis. Methods:Totally 102 cases of pregnant women with acute gastritis were randomly as-signed into two groups with 51 cases in each. The control group was treated with marzulene-S granules 30min before meals with warm water,0. 67g,tid,and the observation group was additionally given phloroglucinol injection 80 mg in 0. 9% sodium chloride injections 20ml,ivd,qd. The treatment course was 2 weeks. The clinical curative effect of the two groups was observed before and after the treat-ment,and the symptom score changes and symptom remission time were compared as well. Results:The effective rate of the observa-tion group was significantly higher than that of the control group(94. 12% vs 78. 43%,P<0. 05). After the treatment,the integral of epigastric pain,belching and dry mouth in the observation group was decreased significantly than that before the treatment(P<0. 05), and significantly lower than that in the control group(P<0. 05). The time of abdominal pain was(1. 8+0. 4)d and(3. 9+1. 1)d in the observation group and the control group,respectively,and the difference was statistically significant(P<0. 05). üo serious ad-verse reactions were found in the two groups. Conclusion:Marzulene-S granules combined with phloroglucinol injections in the treat-ment of pregnant women with acute gastritis is effective.

Objective To investigate the prognosis value of pulmonary infection score and acute physiology and chronic health evaluation to acute exacerbation of COPD with type 2 respiratory failure. Methods 112 cases with acute exacer-bation of COPD with type 2 respiratory failure were selected as research subjects in our hospital from February 2011 to February 2014,scores of two scoring methods,actual mortality with different scores of two scoring methods,ROC curve predicted mortality of two scoring methods to chronic obstructive pulmonary disease with acute exacerbation of respira-tory failure of survival group and death group were compared. Results There were no significant differences between the two groups in PaO2、pH、PaCO2 and HCO3-(t=1.742, 1.647, 1.791, 1.082, P>0.05);but the levels of AG and poten-tial HCO3-in death group were higher than those in survival group(t=10.865, 5.993, P<0.05). APACHEⅢ,CPIS scores of survival group significantly were lower than the death group, the differences were statistically significant (t=6.254,4.397,P<0.05). Mortality of APACHⅢ score ≥25 and CPIS score ≥6 were significantly higher than patients with lower than that value, the differences were statistically significant(χ2=10.264,7.351,P<0.05). CPIS predict mortality acute exacerbation of COPD with type 2 respiratory failure, the area under the ROC curve was 0.867,APACHEⅢpredict mortality acute exacerbation of COPD with type 2 respiratory failure, the area under the ROC curve was 0.938. Conclusion CPIS and APACHEⅢ has certain prognostic value to acute exacerbation of COPD and type 2 respiratory failure,but APACHEⅢ resolution are better, it can provide a reference for disease prediction, it is worthy of applica-tion and promotion in clinical.

OBJECTIVE: To improve the quality standard of Folium Mahoniae. METHODS: TLC was used for qualitative identification. The contents of moisture, ash and ethanol extract were determined. The content of berberine hydrochloride was determined by HPLC. The determination was performed on WondaSil C18 column with mobile phase consisted of acetonitrile-0. 05 mol/L monopotassium phosphate solution (25: 75, V/V) at the flow rate of 1. 0 mL/min. The detection wavelength was 264 nm, column temperature was 30 ℃, and sample size was 10 μL. RESULTS: TLC spots were clear and well-separated. The contents of moisture, total ash, acid-insoluble ash and ethanol extract were 3. 92％-7. 03％, 3. 65％-6. 95％, 0. 05％-1. 03％ and 10. 87％-33. 14％, respectively. The linear range of berberine hydrochloride was 0. 183-0. 915 μg(r=0. 999 9); quantitation limit and detection limit was 0. 143, 0. 095 μg, respectively. RSDs of precision, stability and reproducibility tests were lower than 2. 0％ (n=6); recovery was 95. 21％ -103. 10％ (RSD = 2. 95％, n=6). CONCLUSIONS: The content of moisture, total ash and acid-insoluble ash of medicinal materials is not exceed 8. 0％, 6. 0％ and 0. 4％, respectively. The content of ethanol extract and berberine hydrochloride is not less than 16. 0％ and 1. 0％, respectively. Established standard can be used for quality control of Folium Mahoniae.

Objective To investigate the predictive value of diffusion-weighted (DW) magnetic resonance imaging (MRI) for invasiveness of hilar cholangiocarcinoma (HC).Methods The retrospective casecontrol study was conducted.The clinicopathological data of 65 HC patients who were admitted to the Sun Yat-sen Memorial Hospital from January 2012 to November 2017 were collected.Patients received DW MRI before treatment,and 2 senior imaging doctors analyzed imaging data and measured the apparent diffusion coefficient (ADC) for the primary lesions of HC.Observation indicators:(1) MRI situations of HC;(2) relationship between ADC and clinicopathological factors;(3) receiver operator characteristic (ROC) curve analysis;(4) treatment and follow-up situations.According to patients' conditions,treatment plans were done within 2 weeks after MRI and patients underwent radical resection of HC.Follow-up using telephone interview was performed to detect tumor recurrence up to December 2017.Measurement data with normal distribution were represented as (x)±s,and comparisons between group and among group were respectively analyzed using the t test and one-way ANOVA.Spearman's rank correlation was performed to analyze the relationship between ADC and clinicopathological factors.ROC curves assessed the diagnostic efficiency of ADC.Results (1) MRI situations of HC:MRI and magnetic resonanced cholangio-pancreatography (MRCP) in 65 patients showed varying degrees of soft rattan-like dilations of intrahepatic bile ducts and truncation signs of bile tracts in hepatic port.Of 65 patients,tumors in 23,7 and 35 patients were respectively pedunculated type,polypoid type and infiltrating type.The pedunculated-type lesions of 23 patients presented as low signal on T1WI and slightly high signal on T2WI;after enhanced scans of MRI,pedunculated-type lesions of 7 patients demonstrated moderate homogenous enhancement in 3 patients,ring-like enhancement with internal liquefaction necrosis in 10 patients and moderate heterogeneous enhancement in 10 patients,respectively.The polypoid-type lesions presented as low signal on T1WI and high signal on T2WI,and moderate homogenous enhancement by enhanced scans of MRI.There were varying degrees of bile duct wall thickness and irregular nodules in the infiltrating-type lesions of 35 patients,showing moderate enhancement by enhanced scans of MRI.All the lesions of 65 patients using DW MRI demonstrated restricted diffusion,showing a clear boundary between lesions and normal surrounding bile ducts or liver tissues;heterogeneous enhancement lesions by MRI scans presented as heterogeneously high signal on DWI and heterogeneously low signal on ADC map,and necrotic area of lesions showed low signal on DWI;homogenous enhancement by MRI scans presented as homogenously high signal on DWI and homogenously low signal on ADC map.(2) Relationship between ADC and clinicopathological factors:ADC was respectively (1.382±0.165)× 10-3 mm2/s,(1.343±0.138)× 10-3 mm2/s,(1.291-±0.226)×10-3 mm2/s,(1.111±0.243)×10-3 mm2/s in stage Ⅰ,Ⅱ,Ⅲ and Ⅳ (TNM staging) and (1.441± 0.355) × 10-3 mm2/s,(1.226 ± 0.177) × 10-3 mm2/s,(1.061 ± 0.228) × 10-3 mm2/s in highdifferentiated,moderate-differentiated and low-differentiated tumors (pathological grading) and (1.403±0.176)× 10-3 mm2/s,(1.121±0.238)× 10-3 mm2/s in Ki-67 score ≤ 10％ and ＞ 10％ and (1.115±0.241)× 10-3 mm2/s,(1.347±0.174)× 10-3 mm2/s in HC patients with and without lymph node metastasis,with statistically significant differences in the above indicators (F =4.158,9.866,t =11.607,13.464,P＜0.05).Results of Spearman's rank correlation analysis showed that ADC had a negative correlation with TNM staging,pathological grading and Ki-67 score (r=-0.532,-0.522,-0.409,P＜0.05).(3) ROC curve analysis:using 1.225×10-3 mm2/s as a critical value of ADC,the sensitivity and specificity of ADC in the diagnosis of stage Ⅰ-Ⅱ HC and stage Ⅲ-Ⅳ HC were 70.5％ and 81.0％,and area under ROC curve was 0.705 (95％CI:0.62-0.84,P＜0.05).Using 1.100×10-3 mm2/s as a critical value of ADC,the sensitivity and specificity of ADC in the diagnosis of lowdifferentiated HC and moderate-and high-differentiated HC were 88.2％ and 64.3％,and area under ROC curve was 0.814 [95％ confidence interval (CI):0.69-0.90,P＜0.05].Using 1.243×10-3 mm2/s as a critical value of ADC,the sensitivity and specificity of ADC in the diagnosis of Ki-67 score ≤ 10％ and ＞ 10％ were 66.7％ and 75.0％,and area under ROC curve was 0.783 (95％CI:0.62-0.90,P＜0.05).Using 1.222×10-3 mm2/s as a critical value of ADC,the sensitivity and specificity of ADC in the diagnosis of lymph node metastasis were 91.3％ and 71.4％,and area under ROC curve was 0.873 (95％CI:0.76-0.94,P＜0.05).(4) Treatment and followup situations:65 patients underwent successful radical resection of HC.Thirty-three patients were followed up for 1-24 months.Of 33 patients,5 had tumor recurrence within 6 months postoperatively,including 4 with ADC ＜ 1.100× 10-3 mm2/s,13 had tumor recurrence after 6 months postoperatively,and 15 didn't have tumor recurrence or metastasis,including 1 with ADC ＜ 1.100× 10-3 mm2/s.Conclusions There are different ADC in differentTNM staging,pathological grading,Ki-67 score and with or without lymph node metastasis of HC.ADC of DWMRI can be used as a preoperative imaging predictor for invasiveness of HC.

Objective:This study compares the magnetic resonance imaging (MRI) appearance of two types of breast tissue markers to investigate the appropriate clinical application of the markers.Methods:Breast MRI of 69 patients (78 masses) with breast tissue markers had been placed were analyzed retrospectively from November 2015 to August 2018 in our hospital. The sizes and shapes of breast tissue markers were assessed in axial fat-suppressed T2-weighted images, T1-weighted images and contrast-enhanced T1-weighed images.Results:The length of the coil nickel-free stainless steel markers were greater than ribbon titanium markers, with statistical difference in fat-suppressed T2-weighted images ( P=0.039). In contrast-enhanced T1-weighted images, all coil nickel-free stainless steel markers showed >6 mm diameter and round shape, and ribbon titanium markers showed >6 mm diameter ( n=20) or ≤6 mm diameter ( n=8), and round ( n=20), dot ( n=7) or band ( n=1) shapes. The categories of sizes and shapes in two types of breast tissue markers both had statistical significance ( P<0.001, P<0.001). Conclusions:Small breast lesions with breast tissue markers are not suitable for MRI evaluation. The artifact of ribbon titanium markers is smaller than coil nickel-free stainless steel markers, so they have less impact for lesions. The choice of the breast tissue markers and image evaluation methods should depend on the different clinical conditions.

OBJECTIVETo investigate the effect of co-expression of bone morphogenetic protein 2 (BMP2) and Sox9 on chondrogenic differentiation of mesenchymal stem cells (MSCs) in vitro and provide experimental evidence for tissue engineering of cartilage.

METHODSMouse embryonic bone marrow MSC C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP2, Sox9 and green fluorescent protein (GFP) for 3-14 days, with cells infected with the adenovirus carrying GFP gene as the control. The mRNA expression of the markers of chondrogenic differentiation, including collagen type II (Col2a1), aggrecan (ACAN), and collagen type X (Col10a1), were determined by real-time PCR. Alcian blue staining was used for quantitative analysis of sulfated glycosaminoglycan in the cellular matrix. The expression of Col2a1 protein was assayed by immunohistochemical staining and Western blot analysis.

RESULTSAdenovirus-mediated BMP2 expression induced chondrogenic differentiation of C3H10T1/2 cells. Overexpression of Sox9 effectively enhanced BMP2-induced expression of the chondrogenic markers Col2a1, aggrecan and Col10a1 mRNAs, and promoted the synthesis of sulfated glycosaminoglycan and Col2a1 protein in C3H10T1/2 cells.

CONCLUSIONCo-expression of BMP2 and Sox9 can promote chondrogenic differentiation of MSCs in vitro, which provides a new strategy for tissue engineering of cartilage.

Animals ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Cartilage ; cytology ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

BACKGROUND: Dysfunction of the gap junction channel protein connexin 43 (Cx43) contributes to myocardial ischemia/reperfusion (I/R)-induced ventricular arrhythmias. Cx43 can be regulated by small ubiquitin-like modifier (SUMO) modification. Protein inhibitor of activated STAT Y (PIASy) is an E3 SUMO ligase for its target proteins. However, whether Cx43 is a target protein of PIASy and whether Cx43 SUMOylation plays a role in I/R-induced arrhythmias are largely unknown. METHODS: Male Sprague-Dawley rats were infected with PIASy short hairpin ribonucleic acid (shRNA) using recombinant adeno-associated virus subtype 9 (rAAV9). Two weeks later, the rats were subjected to 45 min of left coronary artery occlusion followed by 2 h reperfusion. Electrocardiogram was recorded to assess arrhythmias. Rat ventricular tissues were collected for molecular biological measurements. RESULTS: Following 45 min of ischemia, QRS duration and QTc intervals statistically significantly increased, but these values decreased after transfecting PIASy shRNA. PIASy downregulation ameliorated ventricular arrhythmias induced by myocardial I/R, as evidenced by the decreased incidence of ventricular tachycardia and ventricular fibrillation, and reduced arrythmia score. In addition, myocardial I/R statistically significantly induced PIASy expression and Cx43 SUMOylation, accompanied by reduced Cx43 phosphorylation and plakophilin 2 (PKP2) expression. Moreover, PIASy downregulation remarkably reduced Cx43 SUMOylation, accompanied by increased Cx43 phosphorylation and PKP2 expression after I/R. CONCLUSION PIASy downregulation inhibited Cx43 SUMOylation and increased PKP2 expression, thereby improving ventricular arrhythmias in ischemic/reperfused rats heart.
Rats ; Male ; Animals ; Myocardial Reperfusion Injury/metabolism* ; Connexin 43/genetics* ; Sumoylation ; Down-Regulation ; Rats, Sprague-Dawley ; Arrhythmias, Cardiac/drug therapy* ; Myocardial Ischemia/metabolism* ; RNA, Small Interfering/metabolism*