Objective To investigate the effects of IGF-Ⅱ on adriamycin-induced apoptosis of hepatocellular carcinoma cell line(HepG2).Methods Cultured HepG2 cells were divided into:①group A:control group;②group B:ADM group(200 ng/ml);③group C:2 ng/ml IGF-Ⅱ+200 ns/ml ADM group;④group D:20 ng/ml IGF-Ⅱ+200 ng/ml ADM group;⑤group E:200 ng/ml IGF-Ⅱ+200 ng/ml ADM group. After HepG2 were treated for 48 h,MTT colorimetry was performed to determine the cell viability,Flow cytometry was used to detect the cell apoptosis rate,and Weastern-blot was performed to evaluate the expression of survivin. Results The cell viability of group A、B、C、D、E was respectively 0.568±0.025、0.201±0.020、0.232±0.027、0.268±0.013 and 0.304±0.019;The cell apoptosis rate of group A、B、C、D;E was respectively 6.9％±1.3％、35.4％±2.1％、31.2％±2.2％、26.4％±1.7％and 21.7％±1.9％;Survivinβ-actin of group A、B、C、D、E was respectively 0.527±0.039、0.147±0.081、0.311±0.069、0.421±0.033 and 0.469±0.031.The cell viability in IGF-Ⅱ+ADM group was better than ADM group(P＜0.01),the cell apoptosis rate in IGF-Ⅱ+ADM group was significantly lower than ADM group(P＜0.01),the expression level of survivin in IGF-Ⅱ +ADM group was significantly higher than ADM group(P＜0.01). Conclusions IGF-Ⅱ inhibits ADM-induced apoptosis of HepG2 cells,probably by up-regulating cell's survivin expression.

ObjectiveTo study the chromosome genetic changes in hepatocellular carcinoma (HCC) with double exposure to hepatitis B virus/aflatoxin B1 (HBV/AFB1) in Guangxi.Method Differences in genomic alterations in 32 patients with HCC were analyzed using comparative genomic hybridization(CGH).Results(1) The majority of chromosome copy number in the 32 HCC samples had varying degrees of change.The amplification of chromosome regions were 1q,7q,8q,with the high frequency regions being 1q,8q.The deletion of chromosome regions were 1p,4q,8p,9p,13q,14q,16p,16q,17p,18q,19p,Y,with the high frequency regions being 1p,4q,8p,16q,17p,19p;(2) There were also some high copy number amplification or deletion of small regions,such as 2p25.1-p25.2,3q22.3-q23,7p14.1-p14.3,and 9p13.2-9p21; (3) Hierarchical clustering analysis showed that the rate of deletion of chromosome 13q decreased progressively in the following 4 groups:-HBsAg(+)/AFB1 (+),HBsAg(+)/AFB1 (-),HBsAg( - )/AFB1 ( + ),and HBsAg( - )/AFB1 (-) (x2=6.452,P＜0.05).4p was found mainly to be amplified in the HBsAg(+)/AFB1(-)group,but it was mainly deleted in the HBsAg(-)/AFB1(+),and HBsAg( - )/AFB1(-) groups.19q was found mainly to be amplified in the HBsAg(+)/AFB1(+) group,but it was mainly deleted in the HBsAg(-)/AFB1(+),and HBsAg(-)/AFB1(-) groups.ConclusionThe chromosome genetic changes of HCC in Guangxi showed multiplicity.The deletion of chromosome 19p,2p25.1-25.2,3q22.3-q23,7p14.1-p14.3 and amplification of chromosome 9p13.2-9p21 are probably unique genetic characteristics of HCC in this region.The combined effects of Hepatitis B virus and aflatoxin B1 may contribute to deletion of chromosome 13q of HCC in Guangxi.

Objective To compare clinicopathological characteristics and prognosis between triple-negative and HER-2-overexpressed breast cancer patients.Methods From Jan.1997 to Jan.2007,data of 725 cases of primary breast cancer patients undergoing surgical treatment were analyzed.The patients were classified into triple-negative and HER-2-overexpressed phenotypes based on immunohistochemistry results.The clinicopathological characteristics and survival of the 2 groups were compared.Results Of the 725 cases,triple-negative and HER-2-overexpressed phenotypes accounted for 12.29%and 24.96%respectively.Compared with HER-2-overexpressed group,the triple-negative group had significantly higher proportion of familial history of malignancy (18.4% vs 5.5%,P=0.001)and higher proportion of patients'histological grade reaching grade 3(54.0% vs 42.0%,P=0.01).There were more lymph node metastasis in triple-negative group than that in HER-2-overexpression group(N1+N2+N3:74.7%vs 64.6%,P=0.045).The recurrence and metastasis rate within 2 years and brain metastasis rate in triple-negative group were higher than those in HER-2-overexpressed group (25.3%,8.0%vs 8.8%,2.2%,respectively,P<0.05).The 5-year disease-free survival rate in triple-negative group was significantly lower than that in HER-2-overexpressed group(55.6%vs 69.8%,P=0.041).There was no statistical difference between the 2 groups in terms of age,menopausal status,tumor size,pathological stage,surgcal procedure,pathological type,adjuvant radio-chemotherapy,the proportion of metastasis to liver,lung and bone as well as overall survival rate(P>0.05).Conclusion Compared to HER-2-overexpressed group,patients with triple-negative breast cancer show higher rate of familial history of malignancy,and they are associated with higher histological grade and poor prognosis.

Objective To investigate the correlation between Young′s elastic modulus (EI) using shear wave elastography (SWE) and liver pathology .Methods Liver biopsy was performed on 231 patients with chronic hepatitis B (CHB) under supersonic guidance ,and SWE with EI of liver was obtained concurrently .The correlation between measured liver stiffness and pathology was analyzed by using the liver pathology as golden standards .One‐way analysis of variance and Spearman rank correlation analysis were performed for the comparison between groups and correlation between two variables , respectively .Receiver operating characteristic (ROC) curve was used to explore the predictive value of shear modulus for the liver inflammation grades and fibrosis stages .Results The EI medians of different liver inflammation grades were 6 .78 kPa (G1) ,7 .30 kPa (G2) ,9 .93 kPa (G3) and 14 .93 kPa (G4) , respectively ,which were statistically different (H=55 .19 ,P<0 .01) .And EI medians of various fibrosis stages were 6 .62 kPa (S0 -S1) ,7 .15 kPa (S2) ,9 .78 kPa (S3) and 14 .62 kPa (S4) ,respectively , which were also significantly different (H=62 .14 ,P<0 .01) .EI was positively correlated with both liver inflammation grades (r=0 .454 6 ,P<0 .01) and liver fibrosis stages (r=0 .505 6 ,P<0 .01) .The areas under the ROC for G≥2 ,G≥3 and G=4 were 0 .68 (95% CI:0 .61 -0 .75) ,0 .77 (95% CI:0 .70 -0 .84) and 0 .85 (95% CI:0 .77-0 .92) ,respectively .The areas under the ROC for S≥2 ,S≥3 and S=4 w ere 0 .73 (95% C I:0 .66 -0 .79 ) ,0 .78 (95% C I:0 .72 -0 .85 ) and 0 .83 (95% C I:0 .75 -0 .91 ) , respectively .Conclusion The EI measured by SWE is correlated with liver pathology of CHB patients , which may be used to dynamically monitor the progress of liver fibrosis .

To identify the related substances in pioglitazone hydrochloride by hyphenated LC-MS techniques,an Ultimate XB-C18 (250 mm × 4.6 mm,5 μm) column was used for separation of the related substances with methanol and 0.1％ ammonium acetate buffer as the mobile phases in gradient elution.Electrospray positive ionization high resolution TOF/MS was used for the determination of the accurate mass and elemental composition of parent [M + H] + ions of the related substances,and triple quadrupole tandem mass spectrometry was employed for the product mass spectra determination.Eleven major related substances were detected and identified to be one synthesis intermediate,six by-products and four degradation products,by using LC-MS determination,spectra elucidation,and further synthetic process and stress degradation mechanisms analysis.The results are useful for pioglitazone hydrochloride manufacturing processes optimization and quality control.

Objective To investigate the causes and managements of postoperative complications of proximal femoral nail anti-rotation(PFNA)in treating intertrochanteric fractures. Methods A total of 155 patients with intertrochanteric fractures were treated with PFNA from January 2011 to December 2017.Causes and managements of postoperative complications were analyzed and discussed. Results At an average follow-up of 48 months(range ,12~72 months) ,the rate of good postoperative recovery in hip function was 96.0% . Among 155 patients ,the internal fixation-associated postoperative complications occurred in 15(9.7% )patients.Details were as follows :7 cases had spiral blade breakage into the femoral head or inwardly ,in whom 5 cases received artificial joint replacement whereafter.5 cases had coxa vara ,mainly in patients with obvious posterior fracture of the femoral intertrochanter with obviously bone fragments.3 cases had delayed fracture healing due to the fracture damaging calcar femorale ,or extensive soft tissue dissection during the operation on small trochanter. The Harris function score was 82 points. Conclusions The location of the PFNA spiral blade ,as an important factor ,is correlated with postoperative complications ,while severe osteoporosis ,unstable fractures ,and poor fracture reduction increase the incidence of surgical complications.

Objective:To explore the effect of Xuanbai-Chengqi Decoction combined with conventional western medicine therapy for the coma patients with acute cerebral infarction. Methods:A total of 72 patients with acute cerebral infarction in Huaibei Hospital of Traditional Chinese Medicine from March 2019 to January 2020 were randomly divided into two groups with 36 in each group. The control group was treated with conventional western medicine therapy, and the treatment group was given Xuanbai-Chengqi Decoction by nasal feeding on the basis of the control group. Both groups were treated for 7 days. The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate the degree of neurological deficit, and the Full Outline of Unresponsiveness Scale (FOUR) was used to evaluate the degree of consciousness disorder of patients. Three-dimensional reconstruction of head CT was performed to identify and mark the edema area. The levels of serum high-sensitivity C-reactive protein (hs-CRP), homocysteine (Hcy) and vascular endothelin-1 (ET-1) were measured by double antibody sandwich enzyme-linked immunosorbent assay. The adverse events during treatment were observed and the clinical effective rate was evaluated. Results:The total effective rate was 100% (36/36) in the treatment group and 86.1% (35/36) in the control group, and the difference between the two groups was statistically significant ( Z=-0.242, P=0.015). On the 3rd and 7th day after treatment, the NIHSS scores of the treatment group were significantly lower than those in the control group ( t values were 26.567 and 17.982, all Ps<0.01). On the 3rd and 7th day after treatment, the eye opening response, motor response, brainstem response, brain stem response and total scores (3 days after treatment, t=15.235 , 14.892, 18.452, 11.232, 16.235; 7 days after treatment, t=19.5 68, 16.232, 10.356, 9.546, 11.098) of the treatment group were significantly lower than those in the control group. The levels of serum hs-CRP, Hcy, ET-1 and CT threshold of brain edema in the treatment group were significantly lower than those in the control group after treatment ( t=22.352, 17.789, 11.908 and 19.652, all Ps<0.01). There were no adverse drug reactions, no abnormal changes in blood routine tests, liver function and electrocardiogram in both groups. Conclusion:The Xuanbai-Chengqi Decoction combined with conventional western medicine therapy can improve the neurological function and promote awakening of coma patients with acute cerebral infarction, which may be related to reducing the levels of inflammatory cytokines related to hs-CRP, Hcy and ET-1, improving microcirculation and relieving brain edema.

Objective:To establish a disk (CD) microfluidic chip detection platform for the rapid detection of CALR-1 and CALR-2 mutations in patients with cerebral infarction, and summarize its clinical application value.Methods:Based on microfluidic technology and loop mediated isothermal amplification technology, a CD microfluidic chip detection platform for simultaneous detection of CALR-1 and CALR-2 gene mutations were established, and the sensitivity, specificity, repeatability and accuracy of the platform were verified. A total of 124 patients with cerebral infarction treated in Huashan Hospital, Shanghai Medical College, Fudan University from November 2019 to March 2021 were prospectively selected into the experimental group; and 80 healthy subjects were included in the control group. The CALR-1 and CALR-2 gene mutations in anticoagulant peripheral blood samples were detected by the CD microfluidic chip. Each chip could detect 4 samples at the same time and synchronously detect 3 indexes of each sample. The detection results could be obtained after isothermal amplification for 40 min. At the same time, sequencing method was used to verify the test results, and the consistency of the results of the two detection methods was compared.Results:Using this CD microfluidic chip platform, the synchronous amplification of 3 indexes in the sample could be completed within 40 min without the need of thermal circulation, and the whole detection process of the sample could be completed within 60 min. For samples with a high concentration of target nucleic acid, typical positive signals could be visualized after amplification for 10 min, and the test results would be available within 30 minutes after receiving the samples. The detection sensitivity of CD microfluidic chip method for CALR-1 and CALR-2 mutation load concentration was 1.0% and 0.5% respectively. Nonspecific amplification was not observed for the non-target nucleic acid samples, indicating the high specificity of this method. The coincidence rates of intra and inter batch repeatability were 100% (20/20) respectively. Two samples with CALR gene mutation were found in the cerebral infarction group, both of which were CALR-1 mutations (L367fs*46). There was no CALR-1 or CALR-2 mutation in the control group. The detection results of CD microfluidic chip method were completely consistent with the sequencing verification results (100% [204/204]).Conclusions:The CD microfluidic chip method could be used for the detection of CALR-1 and CALR-2 gene mutations in clinical samples of patients with cerebral infarction. This method has the advantages of high detection sensitivity, good detection specificity, fast detection speed and high detection flux, which is helpful to clarify the etiology of patients with cerebral infarction.