Objective To establish the insulin resistant HepG2(HepG2/IR)cells model,and investigate the relationship of insu-lin resistance and reduced susceptibility to chemotherapy in hepatoma cells and its mechanism.Methods HepG2 cells were cultured in medium containing 0.5μmol/L insulin for different hours to induce insulin resistance.Glucose consumption of HepG2/IR cells were measured by Hitachi 7600 automatic biochemical analyzer.The cis-dichlorodiamineplatinum(DDP)sensitivity of the HepG2 and HepG2/IR cells were determined by MTT assay,the Annexin Ⅴ/PI assay was adopted to measure the apoptosis rate.In addi-tion,real-time PCR,flow cytometry (FCM)and Western-Blot were employed to detect the mRNA and protein levels of insulin re-ceptor(InsR)and endoplasmic reticulum chaperonin 78(GRP78).Results The glucose consumption decreased and expression of In-sR was down-regulated in HepG2/IR cells.The HepG2/IR cells had reduced sensitivity to DDP (P<0.05 ).The IC50 s of the HepG2/IR cells treated by DDP for 48 h and 72 h were 158.8% and 165.9% of HepG2 cells respectively,while the apoptosis rate was 50.29% lower.The mRNA and protein level of GRP78 in HepG2/IR cells were 2.12 and 2.27 times of that in HepG2 cells. Conclusion The stable HepG2/IR cells showed stronger resistance to DDP were established from HepG2 cell induced with insulin, and its mechanism may be related to the increased expression of GRP78.

BACKGROUND: Migu tablet, a Chinese drug for kidney invigorating, is effective on preventing and treating osteoporosis, but the concrete mechanism of pharmacology is still not clear. Transforming growth factor-β1(TGF-β1) is an important cytokine, which can regulate bone resorption and formation.OBJECTIVE: To investigate the effect of kidney invigorating recipe on mRNA expression of TGF-β1 of osteoblast.DESIGN: A completely randomized controlled study was conducted.SETTING: Department of Traumatic Orthopedics, Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: This experiment was conducted at the laboratory for bone metabolism of integration of Chinese and western medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from May 2003 to April 2004. Experimental rats: Totally 16 newborn SD rats of clean degree were involved. Experimental drug: Medical liquor of Migu tablet was prepared in the Department of Traumatic Orthopedics,Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. The subscription was mainly composed of Chinese herbs such as Herba Epimedii, Cortex Eucommiae, Semen Juglandis,Radix Rehmanniae, Radix Achyranthis Bidentatae. and so on. Positive control drug: which was recombinant basic fibroblastic growth factor (rbFGF), was purchased from Beijing Banding Company. Negative control group was subdivided into negative control of probe and antibody METHODS: 100,1 000,5 000,10 000 mg/L Chinese herb Migu tablet liquor for kidney invigorating and positive control drug 5 μg/L rbFGF were added into the osteoblasts of cranial bones of newly born SD rats separated and cultured in vitro. 24 hours later, nuclear acid molecular in situ hybridization of osteoblasts were analyzed by self-made digoxin-labeled TGF-β1 cDNA probe . The mean absorbance of positive particles representing the mRNA expression of TGF-β1. A total of 40 osteoblasts were randomly chosen from each group under 200-fold amplification. The average absorbance of hybridized particles of the cells was measured with TJTY-300 automatic image analyzer.MAIN OUTCOME MEASURES: mRNA expression of TGF-β1 in osteoblasts of each group.RESULTS: Automatic image analyzer showed that the TGF-β1 mRNA expressions of Migu liquor groups whose concentration were 5 000 mg/L and 10 000 mg/L were respectively 1.18 times and 1.30 times that of control group, with a significant difference. [The mean absorbance's of hybridized particles of the cells in the 5 000,10 000 mg/L Migu liquor groups and negative control were 0.213 67±0.015 00,0.237 03±0.021 73,0.181 27±0.015 28 ,respectively, P ＜ 0.05 and P ＜ 0.01].Although the mean absorbance ( 0.254 45±0.020 81)of the hybridized particles of the cells in the 5 μg/L recombinant rbFGF was higher than those of 5 000,10 000 mg/L Migu liquor groups, but there was no significant difference(P ＞ 0.05).CONCLUSION: Migu tablet for kidney invigorating can stimulate the secretion and synthesis of TGF-β1 in osteoblasts, thus promote bone formation and inhibit bone resorption.

Purpose The one-step examination of myocardial perfusion imaging (MPI) combined with coronary artery calcium score (CACS) can obtain both coronary functional information and anatomical information simultaneously, this paper aims to evaluate the value of the one-step examination of MPI combined with CACS for detecting coronary artery disease (CAD). Materials and Methods 188 cases who underwent one-step examination of MPI combined with CACS and invasive coronary angiography (ICA) because of chest tightness, chest pain with suspected coronary artery disease were analyzed retrospectively, with the results of ICA used asgold standard, the diagnostic efficacy of MPI, CACS and one-step examination with combination of the two techniques for CAD was investigated. Results ①Pre-test probability of CAD was intermediate in 79.8%(150/188), and high in 20.2%(38/188) cases. Seventy-three cases were confirmed as CAD and 115 of 188 patients were negative according to ICA.②The sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) for the diagnosis of CAD by MPI were 65.8%, 75.7%, 71.8%, 63.1%and 77.7%, respectively. ③ The CACS of CAD group was significantly higher than the non-CAD group (494.96±99.60 vs. 38.15±16.03, P<0.05). According to the features of the ROC curve, the best threshold for the diagnosis of CAD with CACS was 96.45, with CACS≥96.45 as the positive standard in diagnosis of CAD, the sensitivity, specificity, accuracy, PPV and NPV for the diagnosis of CAD by CACS were 60.3%, 93.9%, 80.8%, 86.3%and 78.8%, respectively. ④ The sensitivity of MPI combined with CACS were significantly higher than MPI (80.8% vs. 65.8%, P<0.05), while the specificity (71.3% vs.75.7%, P>0.05) and accuracy (75.0% vs. 71.8%, P>0.05) showed no statistically significant difference; the sensitivity of MPI combined with CACS were significantly higher than CACS (80.8%vs. 60.3%, P<0.05), while the specificity was lower than CACS (71.3%vs. 93.9%, P<0.05) and the accuracy showed no statistically significant difference (75.0%vs. 80.8%, P>0.05). Conclusion The one-step examination of MPI combined with CACS can reduce coronary heart disease misdiagnosis, improve the diagnostic sensitivity of CAD compared with the MPI or CACS, with high application value for the diagnosis of CAD, especially in moderate risk groups.

Objective To evaluate the additional value of CACS in detection of coronary artery disease (CAD) with MPI.Methods A total of 188 suspected CAD patients (128 males,60 females;average age (61.93±9.16) years) who underwent one-step examination of MPI combined with CACS from December 2012 to August 2014 were enrolled in this retrospective study.According to the gold standard of CAG,the diagnostic efficacy of MPI was calculated.ROC analysis was performed to determine the optimal CACS threshold for the detection of CAD.Mann-Whitney u test and x2 test were used for statistical analysis.Results (1) Seventy-three CAD cases were confirmed (≥ 50％ stenosis) among 188 patients.The sensitivity,specificity,accuracy for CAD diagnosis with MPI were 65.8％ (48/73),75.7％ (87/115),71.8％ (135/188),respectively.Twenty-five CAD patients had negative findings with MPI,including 2 with LM disease,4 with three-vessel disease (LAD±LCX±RCA,3-VD),3 with 2-VD,16 cases with 1-VD.Among them 13 cases (52.0％,13/25) had intermediate lesions of 1-VD (50％ ≤ stenosis＜70％).(2) The CACS of CAD group was significantly higher than that of non-CAD group (172.40(19.25,516.45) vs 0;z=-8.465,P＜0.001).According to the ROC analysis,95.1 was the optimal CACS cutoff to detect CAD patients.Combining MPI with CACS (at cutoff of 95.1) improved the sensitivity of MPI (80.8％,59/73;x2 =4.233,P＜0.05) for the detection of CAD,with no significant decrease in specificity and accuracy (71.3％,82/115;75.0％,141/188;x2 values:0.558 and 0.490,both P＞0.05).(3) Of the 25 CAD patients with negative MPI results,11(44.0％,11/25) showed abnormal CACS(CACS≥95.1),consisted of 2 cases of LM disease,4 cases of 3-VD,2 cases of 2-VD,3 cases of 1-VD.Diagnosis was corrected by CACS in 8/9 cases of severe CAD (LM CAD or multivessel disease) which were missed by MPI.Conclusion CACS could offer additional information for MPI in detection of suspected CAD patients,which can improve the sensitivity of MPI for diagnosing CAD,especially for severe CAD with LM lesions or multivessel CAD.

The clinical data of a case of paroxysmal extreme pain disorder(PEPD) in Guangdong 999 Brain Hospital were retrospectively analyzed.The male patient, age of first examination was 7 months, began to have recurrent tonic accompanied by facial redness or cyanosis at 5 months after birth.The patient was diagnosed with epilepsy.The oral solution of sodium valproate and Levetiracetam were not effective.The video electroencephalogram examination displayed that, when the patient had tonic and bradycardia, the synchro electroencephalogram did not show epileptic discharge, so the patient was considered to have non-epileptic tonic.Genetic examination suggested that SCN9A gene mutation of c. 5240T >C resulted in amino acid changes: Val1747Ala.Combined with the skin changes, the patient was diagnosed as PEPD caused by SCN9A gene mutation.After the treatment with Carbamazepine, the patient′s abnormal skin changed and his-epileptic tonic disappeared, and his condition improved significantly.The early stage of PEPD can be mainly manifested as non-epileptic tonic.It is easy to be misdiagnosed as epilepsy, so the patient′s characteristic skin changes should be noticed, and genetic examination is also helpful in the diagnosis of the disease.

Background and purpose:The recurrence and metastasis of gastric cancer seriously affect survival in patients.SOX gene as a regulatory factor of the classical Wnt pathway, may play an important role in the process. This study was to explore the expression of stem cell marker SOX-2 and β-catenin in gastric cancer and to analyze the relationship with recurrence and metastasis after operation.Methods:Immunohistochemistry was used to detect the expression of SOX-2 and β-catenin in 71 tumor samples from 71 cases after surgery for gastric cancer. The correlation between SOX-2 and β-catenin expression and the clinicopathological characteristics of gastric cancer and disease-free survival was analyzed.Results:The SOX-2 protein expression was associated with metastasis, lymph node inifltration or differentiation (P=0.011,P=0.036,P=0.034) in the 71 gastric cancer, but not with gender, age or T stage. β-catenin expression was correlated with metastasis, lymph node invasion or T stage (P=0.025,P=0.014,P=0.026), but was not related to differentiation, gender or age. The survival analysis showed that SOX-2 and β-catenin expression was closely associated with prognosis of patients, and metastatic rate in positive expression was higher than that in negative expression.Conclusion:The expression of SOX-2 and β-catenin is associated with the development, recurrence, metastasis of gastric cancer and may be used as a useful prognostic parameter to predict overall survival.

Objective:To investigate the clinical features of encephalocraniocutaneous lipomatosis (ECCL).Methods:The clinical characteristics, imaging manifestations and electroencephalogram changes of five patients with ECCL from Guangdong 999 Brain Hospital between December 2016 and February 2019 were collected and analyzed.Results:All five cases showed ocular, skin and central nervous system anomalies. Corneal anomalies were found in five cases, eyelid coloboma in three cases, calcification of the globe in two cases, and choristoma in one case. All five cases presented with naevus psiloliparis, three cases with small nodular skin tags on eyelids, and three cases with café-au-lait spots on the trunk. Dysplasia of the right cerebral hemisphere was observed in all five cases, four cases with enlargement of the right ventricle, three cases with arachnoid cysts, and one case with dysplasia of the corpus callosum. The onset of the seizures of five cases was found within one year old. Spasms were observed in five cases, partial seizure in three cases, and tonic seizure in one case. Five cases were drug-resistant epilepsy. Seizures decreased significantly after adrenocorticotropic hormone treatment in one case and seizures free after surgery in one case. One case had seizure free by corpus callosotomy, but had a relapse after four months. Three cases used ketogenic diet, including one case with epileptic seizure reduction, one case with development progress. All five cases had developmental delays. The exon gene sequencing of four cases was found normal. KRAS gene mutation was found in brain tissue of one patient.Conclusions:ECCL is a rare clinical disease that often involves the nervous system, skin and eyes. The seizures of the patients are often difficult to control and have development delays. Surgery may be necessary to control the seizures. ECCL is thought to be somatic mutations, which are hard to detect in the blood and can be found in affected tissues.

Objective:To explore the optimal acceleration factor and feasibility of the compressed SENSE (CS) technique in non-contrast MR coronary angiography (NMRCA) for clinical practice.Methods:The image data of completed coronary CTA and 3.0 T NMRCA sequence in 31 patients with suspected coronary heart disease were prospectively recruited at Fuyang People′s Hospital from August 2021 to November 2021. NMRCA sequences included conventional SENSE2 sequence and CS sequences with acceleration factors of 4, 5, and 6, respectively. The subjective scores of image quality and the objective scores, the contrast ratios between assessed coronaries and myocardium (CMCR) were compared among the 4 groups using the Friedman and Wilcoxon rank sum test.Results:Compared with the conventional SENSE2 [(343±46)s], the scan time of CS4 (269±36), CS5 (214±29) and CS6 (178±26) s were shortened by 21.5%, 37.5% and 48.0%, respectively. There was a good consistency between the subjective scores of the four groups (Kappa=0.769, 95% Cl 0.738-0.800). There was no significant difference in subjective score and CMCR value between CS4 and SENSE2 ( P>0.05). The coronary artery segments of CS5 and CS6 were significantly different from SENSE2 group ( P<0.05). Conclusions:For 3.0 T NMRCA, CS technology shows high feasibility. The CS4 can reduce imaging time while ensuring high-quality coronary arterial images, which has a well-established clinical application value for NMRCA.

Objective　To explore the relationship between gene mutation types and clinical phenotypes in patients with tuberous sclerosis complex accompanied by epilepsy. Methods　TSC gene was detected in patients with tuberous sclerosis accompanied by epilepsy diagnosed in Guangdong Sanjiu brain hospital from October 2013 to October 2019.The patients with gene positive were genotyped and the clinical data of the patients with gene positive were collected to explore the relationship between different gene mutation types and clinical phenotypes. Results　85 patients were TSC gene positive，of which 34 （40.0%） were TSC1 gene mutation in which 4 （11.8%） were splice mutation，10 （29.4%） were frameshift mutation，4 （11.8%） were nonsense mutation and 16 （47.0%） were missense mutation. 51 （60.0%） were TSC2 gene mutation，in which 3 were splice mutation （5.9%），19 were frameshift mutation （37.3%），1 was nonsense mutation （1.9%），25 were missense mutation （49.0%） as well as 3 were large fragment deletion （5.9%）. The mutation rate of frameshift mutation and missense mutation was higher. The age of onset was divided into ≤ 1 year old，～3 years old，～6 years old，～18 years old and >18 years old. It was found that there were significant differences in TSC1 and TSC2 genes among different age of onset （P<0.05）. At the same time，the incidence of renal disease and mental retardation was statistically significant in TSC1 and TSC2 genes （P<0.05 respectively）. In addition，according to the type of gene mutation，the patients were divided into three groups：frameshift mutation group，missense mutation group and other mutations （including splice mutation，nonsense mutation and large fragment deletion） group. It was found that the incidence of heart disease was significantly different in different gene mutation types （P<0.05 respectively）. Conclusion　There were many different types of TSC1 and TSC2 gene mutation types and clinical phenotypes. The onset age of TSC2 mutation is younger and more prone to have kidney disease and mental retardation. Missense mutations are more likely to develop heart disease. The study of genotype-phenotype relationship can make a preliminary assessment of disease development and prognosis in TSC patients.

Purpose To investigate the clinical value of multi-center digital mammography radiomics nomogram model in predicting triple-negative breast cancer(TNBC).Materials and Methods The digital mammograms of 462 patients with pathologically confirmed breast cancer from November 2016 to March 2022 were retrospectively analyzed,including 243 cases from Yijishan Hospital of Wannan Medical College(institution 1),106 cases from Fuyang People's Hospital(institution 2)and 113 cases from Taihe People's Hospital(institution 3).According to the results of immunohistochemistry,a total of 349 breast cancer patients in institution 1 and institution 2 were randomly divided into the training group(244 cases,including 41 TNBC and 203 non-TNBC)and the validation group(105 cases,including 18 TNBC and 87 non-TNBC)according to the ratio of 7∶3,113 breast cancer patients(24 TNBC and 89 non-TNBC)from institution 3 were included in the external validation group.Comparing the mediolateral oblique and cranial cauda digital mammography images,the mammography imaging with larger lesion areas were selected,and the image segmentation and radiomics feature extraction were performed.The radiomics model was constructed by using Logistic regression.The clinicopathological parameters and radiomics scores were used to construct a nomogram.Receiver operating characteristic and decision curve analysis were used to evaluate the model performance.To compare The predictive performance between the models was compared.Results Finally,four radiomics features closely related to TNBC were selected to construct an radiomics model.The area under the curve,sensitivity and specificity of TNBC predicted by the radiomics model in training group,validation group and external test group were 0.868,90.24%and 72.91%,0.827,72.22%and 75.86%,0.837,70.83%and 78.65%,respectively.The area under the curve,sensitivity and specificity of TNBC predicted by the combined model in the training group,validation group and external test group were 0.903,80.49%and 86.70%,0.890,77.78%and 88.51%,0.870,62.50%and 85.39%,respectively.The combined model was better than the single image omics model in predicting TNBC,and the difference was statistically significant between the training group and the verification group(Z=2.061,2.064,both P<0.05),but not between the external test group(Z=1.223,P=0.221).In three group,decision curve analysis showed that the nomogram predicted a higher net benefit than the radiomics model for triple-negative breast cancer.Conclusion The radiomics model has high diagnostic efficiency in predicting TNBC,and the nomogram model combined with the radiomics score and histological grading can further improve the prediction efficiency.