The tumor microenvironment of hepatocellular carcinoma and its targeting strategy by CAR-T cell immunotherapy.

Hepatocellular carcinoma (HCC) is the major subtype of liver cancer, which ranks sixth in cancer incidence and third in mortality. Although great strides have been made in novel therapy for HCC, such as immunotherapy, the prognosis remains less than satisfactory. Increasing evidence demonstrates that the tumor immune microenvironment (TME) exerts a significant role in the evolution of HCC and has a non-negligible impact on the efficacy of HCC treatment. In the past two decades, the success in hematological malignancies made by chimeric antigen receptor-modified T (CAR-T) cell therapy leveraging it holds great promise for cancer treatment. However, in the face of a hostile TME in solid tumors like HCC, the efficacy of CAR-T cells will be greatly compromised. Here, we provide an overview of TME features in HCC, discuss recent advances and challenges of CAR-T immunotherapy in HCC.

An anoikis-based signature for predicting prognosis in hepatocellular carcinoma with machine learning.

Background: Hepatocellular carcinoma (HCC) is a common malignancy with high mortality worldwide. Despite advancements in diagnosis and treatment in recent years, there is still an urgent unmet need to explore the underlying mechanisms and novel prognostic markers. Anoikis has received considerable attention because of its involvement in the progression of human malignancies. However, the potential mechanism of anoikis-related genes (ANRGs) involvement in HCC progression remains unclear. Methods: We use comprehensive bioinformatics analyses to determine the expression profile of ANRGs and their prognostic implications in HCC. Next, a risk score model was established by least absolute shrinkage and selection operator (Lasso) Cox regression analysis. Then, the prognostic value of the risk score in HCC and its correlation with clinical characteristics of HCC patients were further explored. Additionally, machine learning was utilized to identify the outstanding ANRGs to the risk score. Finally, the protein expression of DAP3 was examined on a tissue microarray (TMA), and the potential mechanisms of DAP3 in HCC was explored. Results: ANRGs were dysregulated in HCC, with a low frequency of somatic mutations and associated with prognosis of HCC patients. Then, nine ANRGs were selected to construct a risk score signature based on the LASSO model. The signature presented a strong ability of risk stratification and prediction for overall survival in HCC patients.Additionally, high risk scores were closely correlated with unfavorable clinical features such as advanced pathological stage, poor histological differentiation and vascular invasion. Moreover, The XGBoost algorithm verified that DAP3 was an important risk score contributor. Further immunohistochemistry determined the elevated expression of DAP3 in HCC tissues compared with nontumor tissues. Finally, functional analyses showed that DAP3 may promote HCC progression through multiple cancer-related pathways and suppress immune infiltration. Conclusion: In conclusion, the anoikis-based signature can be utilized as a novel prognostic biomarker for HCC, and DAP3 may play an important role in the development and progression of HCC.

Effect of compound glycyrrhizin and tacrolimus ointmenta combined with photon therapy on hormone dependence facial dermatitis / 中华医学美学美容杂志

Objective:To observe the clinical curative effect of compound glycyrrhizin tablets and tacrolimus ointmenta combined with photon therapy on hormone dependence facial dermatitis and the safety.Methods:80 cases were randomly divided into two groups: treatment group and control group, 40 each; compound glycyrrhizin tablets and tacrolimus ointmenta were administrated to all the cases in both groups while photon therapy was added to cases in treatment group; a comparative study was made to observe the clinical effect 4 weeks after treatment between the two groups.Results:Four weeks after treatment, the effective rate was 86.67% in treatment group and 63.33% in control group, the difference was of statistical significance ( χ2=4.36, P<0.05); the total scores of symptoms and signs decreased obviously in both groups while the difference between the groups was of statistical significance ( t=3.10, P<0.05); topical medication led to drug adverse reaction in 4 cases in treatment group and 3 cases in control group with the manifestation of tolerable aggravated facial erythema and twinge while the adverse reaction disappeared gradually with the drug application keeping on for about 1 week. Conclusions:Compound glycyrrhizin tablets and tacrolimus ointmenta combined with photon therapy is of definite curative effect on hormone dependence facial dermatitis with good safety.

Ac-SDKP increases α-TAT 1 and promotes the apoptosis in lung fibroblasts and epithelial cells double-stimulated with TGF-ß1 and silica.

Crystalline silica (SiO2) particles have very strong toxicity to the lungs, and silicosis is an excessive pulmonary interstitial remodeling disease that follows persistent SiO2 injury. We showed here that DNA double strand breaks (DSBs) and apoptosis were aggravated during rat silicosis induced by SiO2 exposure. Ac-SDKP attenuates lung parenchymal distortion and collagen deposition, and decreases the expression of γH2AX, p21, and cleaved caspase-3, as well as improves the reduction of pulmonary function caused by silicosis. In vitro, we found an evolution of smooth muscle actin α (α-SMA), collagen type I (Col I) in both A549 and MRC-5 cells in response to transforming growth factor-beta 1 (TGF-ß1) + SiO2. Only A549 cells showed any reduction in the rate of apoptosis induced by the double stimulation, because of the anti-apoptotic effects of TGF-ß1. N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is an anti-fibrotic tetrapeptide. It also has the ability to promote the apoptosis of leukemia cells. However its role in promoting cell apoptosis in silicosis is still unknown. We here found that Ac-SDKP could induce cell apoptosis and inhibit fibrotic response in A549 and MRC-5 cells treated with TGF-ß1 + SiO2, and these effects depended on regulation of α-tubulin acetyltransferase 1 (α-TAT1). These findings suggest that Ac-SDKP may have therapeutic value in the treatment of silicotic fibrosis.

Proteomic profile of TGF-ß1 treated lung fibroblasts identifies novel markers of activated fibroblasts in the silica exposed rat lung.

We performed liquid chromatography-tandem mass spectrometry (LC-MS/MS) on control and TGF-ß1-exposed rat lung fibroblasts to identify proteins differentially expressed between cell populations. A total of 196 proteins were found to be differentially expressed in response to TGF-ß1 treatment. Guided by these results, we next determined whether similar changes in protein expression were detectable in the rat lung after chronic exposure to silica dust. Of the five proteins selected for further analysis, we found that levels of all proteins were markedly increased in the silica-exposed rat lung, including the proteins for the very low density lipoprotein receptor (VLDLR) and the transmembrane (type I) heparin sulfate proteoglycan called syndecan 2 (SDC2). Because VLDLR and SDC2 have not, to our knowledge, been previously linked to the pathobiology of silicosis, we next examined whether knockdown of either gene altered responses to TGF-ß1 in MRC-5 lung fibroblasts. Interestingly, we found knockdown of either VLDLR or SDC2 dramatically reduced collagen production to TGF-ß1, suggesting that both proteins might play a novel role in myofibroblast biology and pathogenesis of silica-induced pulmonary fibrosis. In summary, our findings suggest that performing LC-MS/MS on TGF-ß1 stimulated lung fibroblasts can uncover novel molecular targets of activated myofibroblasts in silica-exposed lung.

Effect of early mobilization on gastrointestinal function and delirium in patients with mechanical ventilation of chronic obstructive pulmonary disease / 中国基层医药

Objective To investigate the effect of early mobilization on gastrointestinal function and delirium in patients with mechanical ventilation of chronic obstructive pulmonary disease (COPD).Methods A total of 87 mechanically ventilated patients with COPD who met the inclusion criteria were selected as subjects from April 2017 to March 2018 in intensive care unit(ICU) of Huxi Hospital Affiliated to Jining Medical College.The patients were divided into treatment group (44 cases) and control group (43 cases) by the random number table method.On the basis of the same conventional treatment,the treatment group received early mobilization.The incidence of acute gastrointestinal injury (AGI),duration of AGI,incidence of delirium,duration of delirium,days of mechanical ventilation and 28-day mortality were compared between the two groups.Results The incidence rate of AGI in the treatment group was lower than that in the control group[40.9％ (18/44) vs.62.7％ (27/43)],and the duration in the treatment group was shortened[(3.95 ± 1.62)d vs.(5.23 ± 2.03)d],and the incidence rate of delirium in the treatment group was lower[54.5％ (24/44) vs.76.7％ (33/43)],the duration of delirium in the treatment group was shortened [(2.36 t 0.9) d vs.(3.25 ± 1.27) d],the mechanical ventilation time in the treatment group was decreased [(6.39 ± 1.76) d vs.(7.56 ± 1.49) d],the differences were statistically significant (x2 =4.17,t =-2.280,x2 =4.744,t =-2.919,-3.358,all P ＜ 0.05).There was no statistically significant difference in the rate of 28-day mortality between the two groups [2.3 ％ (1/43) vs.9.3 ％ (4/43),x2 =-1.984,P ＞ 0.05].Conclusion Early mobilization can reduce the incidence and duration of AGI and delirium in COPD patients with mechanical ventilation,reduce the severity of AGI,and shorten the time of mechanical ventilation.

Effect of high-flow nasal cannula oxygen therapy on diaphragmatic function in patients with acute exacerbation of chronic obstructive pulmonary disease: a prospective randomized controlled trial / 中华危重病急救医学

Objective To investigate the effect of high-flow nasal cannula oxygen therapy (HFNC) on the clinical efficacy and diaphragm function of patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods The patients with mild to moderate AECOPD (clinical classification Ⅰ-Ⅱ) admitted to Huxi Hospital Affiliated to Jining Medical College from January to October in 2018 were enrolled. The patients were divided into HFNC treatment group and routine oxygen therapy control group (each n = 37) by randomly number table method. The two groups were given bronchiectasis drugs, corticosteroids, expectorant, anti-infection treatment, at the same time, the HFNC treatment group was given HFNC with the initial flow rate of 40 L/min. The routine oxygen therapy control group was given low flow oxygen, and the initial flow rate was 3 L/min. General data such as gender, age, clinical grade, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score were recorded. Bedside ultrasound was used to measure the diaphragmatic excursions during quiet breathing (DEq), diaphragmatic excursions during deep breathing (DEd), and diaphragmatic shallow fast breathing index (D-RSBI) before and 2, 24 and 48 hours after treatment in both groups and compared, meanwhile, arterial blood gas analysis was performed, and arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2) were recorded. Results Two patients in the HFNC treatment group withdrew from the study because they could not tolerate HFNC, while other patients were enrolled in the analysis. There was no statistically significant difference in gender, age, proportion of AECOPDⅡ grade or APACHEⅡscore between the two groups, indicating that the general data of the two groups were comparable and balanced. There was no statistically significant difference in DEq, DEd, D-RSBI, PaO2 or PaCO2 before treatment between the two groups. After treatment, DEp in both groups was decreased gradually with time, it was decreased earlier in the HFNC treatment group, and it showed significant difference as compared with that before treatment at 2 hours after treatment (mm: 18.3±3.1 vs. 20.1±4.2, P < 0.01), and it was significantly lower than that in the routine oxygen therapy control group (mm: 18.3±3.1 vs. 20.3±3.7, P < 0.05); DEd was gradually increased in both groups, it was significantly increased in the HFNC treatment group, and it was significantly higher than that in the routine oxygen therapy control group at 24 hours and 48 hours after treatment (mm: 55.2±7.6 vs. 50.8±9.2 at 24 hours, 59.4±7.7 vs. 53.6±9.1 at 48 hours, both P < 0.05); D-RSBI was decreased gradually in both groups, it was decreased earlier and more significant in the HFNC treatment group, and it was significantly lower than that in routine oxygen therapy control group at 24 hours and 48 hours after treatment (times·min-1·mm-1: 0.41±0.13 vs. 0.51±0.20 at 24 hours, 0.31±0.12 vs. 0.43±0.17 at 48 hours, both P < 0.05). After treatment, there was no statistically significant difference in PaO2 or PaCO2 between the two groups. Conclusion HFNC can effectively relieve diaphragm fatigue in patients with mild to moderate AECOPD, but it had no effect on carbon dioxide retention.

Effect of early mobilization on gastrointestinal function and delirium in patients with mechanical ventilation of chronic obstructive pulmonary disease / 中国基层医药

Objective@#To investigate the effect of early mobilization on gastrointestinal function and delirium in patients with mechanical ventilation of chronic obstructive pulmonary disease(COPD).@*Methods@#A total of 87 mechanically ventilated patients with COPD who met the inclusion criteria were selected as subjects from April 2017 to March 2018 in intensive care unit(ICU) of Huxi Hospital Affiliated to Jining Medical College.The patients were divided into treatment group (44 cases) and control group (43 cases) by the random number table method.On the basis of the same conventional treatment, the treatment group received early mobilization.The incidence of acute gastrointestinal injury(AGI), duration of AGI, incidence of delirium, duration of delirium, days of mechanical ventilation and 28-day mortality were compared between the two groups.@*Results@#The incidence rate of AGI in the treatment group was lower than that in the control group[40.9%(18/44) vs. 62.7%(27/43)], and the duration in the treatment group was shortened[(3.95±1.62)d vs. (5.23±2.03)d], and the incidence rate of delirium in the treatment group was lower[54.5%(24/44) vs. 76.7%(33/43)], the duration of delirium in the treatment group was shortened[(2.36±0.9)d vs. (3.25±1.27)d], the mechanical ventilation time in the treatment group was decreased[(6.39±1.76)d vs. (7.56± 1.49)d], the differences were statistically significant(χ2=4.17, t=-2.280, χ2=4.744, t=-2.919, -3.358, all P<0.05). There was no statistically significant difference in the rate of 28-day mortality between the two groups[2.3%(1/43) vs. 9.3%(4/43), χ2=-1.984, P>0.05].@*Conclusion@#Early mobilization can reduce the incidence and duration of AGI and delirium in COPD patients with mechanical ventilation, reduce the severity of AGI, and shorten the time of mechanical ventilation.

A prospective randomized controlled study on the effects of early mobilization therapy on gastrointestinal function and respiratory mechanics in mechanical ventilation patients with chronic obstructive pulmonary disease / 中国医师进修杂志

Objective To investigate the effects of early mobilization therapy on gastrointestinal function and respiratory mechanics in mechanical ventilation patients with chronic obstructive pulmonary disease (COPD). Methods A prospective randomized controlled study was used. Eighty-five patients with COPD who had stable hemodynamics and required invasive mechanical ventilation from January to December 2017 were enrolled. The patients were divided into treatment group with 43 cases and control group with 42 cases according to the random number table method. The patients in both groups received conventional basic treatment, but the treatment group received early mobilization therapy. The conditions of acute gastrointestinal injury (AGI), incidence of ventilator-associated pneumonia (VAP), duration of mechanical ventilation, 28-day mortality and respiratory mechanics indexes before and after treatment including airway resistance (Raw), lung dynamic compliance (Cd), lung static compliance (Cs), intrinsic positive end expiratory pressure (PEEPi) were compared between 2 groups. Results There were no statistical difference in Raw, Cd, Cs and PEEPi before treatment between 2 groups (P＞0.05). The PEEPi and Raw after treatment in treatment group were significantly lower than those in control group:(6.51 ± 1.46) cmH2O (1 cmH2O=0.098 kPa) vs. (7.30 ± 1.61) cmH2O and (19.23 ± 2.62) cmH2O/(L·s) vs. (20.54 ± 2.50) cmH2O/(L·s), the Cs and Cd were significantly higher than those in control group:(53.14 ± 5.07) ml/cmH2O vs. (49.16 ± 5.10) ml/cmH2O and (26.63 ± 3.28) ml/cmH2O vs. (24.54 ± 1.97) ml/cmH2O, and there were statistical differences (P＜0.05 or＜0.01). The incidences of AGI, duration of AGI, duration of mechanical ventilation and incidence of VAP in treatment group were significantly lower than those in control group: 44.2% (19/43) vs. 66.7% (28/42), (3.56 ± 1.22) d vs. (4.26 ± 1.62) d, (6.79 ± 1.92) d vs. (7.64 ± 1.89) d and 2.3% (1/43) vs. 14.3% (6/42), the severity of AGI was significantly lower than that in control group (patients of gradeⅠ,Ⅱ,ⅢandⅣin treatment group were 11, 6, 1 and 1 case respectively, and patients of grade Ⅰ, Ⅱ, Ⅲ and Ⅳ in control group were 7, 12, 8 and 1 case respectively), and there were statistical differences (P＜0.05). There was statistical difference in 28-day mortality between 2 groups (P＞0.05). Conclusions The incidence of AGI in mechanical ventilation patients with COPD is high. Early mobilization therapy can reduce the incidence and duration of AGI in mechanically ventilated patients with COPD, and reduce airway resistance, increase Cd and Cs, reduce PEEPi, improve respiratory function, shorten mechanical ventilation time, and reduce the incidence of VAP, which is worthy of clinical promotion.

Cognitive profile of children with newly onset benign epilepsy with centro-temporal spikes before treatment:a study of computerized cognitive testing in epilepsy / 中华儿科杂志

<p><b>OBJECTIVE</b>Benign epilepsy with centro-temporal spikes (BECTs) is a common idiopathic partial epileptic syndrome in childhood, which often affect the pre-school and school-age children and a considerable proportion have comorbidity including lower academic achievement and cognitive impairment. Few studies involved the psychocognitive assessment in such a drug-treatable epileptic syndrome especially in the newly diagnosed and medications-naive group. This study aimed to investigate the cognitive characteristics of children with newly onset BECTs before treatment.</p><p><b>METHOD</b>Forty-one outpatients with newly diagnosed BECTs who visited the Clinic during the periods from October 2012 to May 2014 before the medications against epilepsy and 41 healthy controls recruited from regular school in Beijing during the period from July 2013 to March 2014, who matched in age and gender underwent battery testing by computerized cognitive testing in epilepsy (CCTE). The BECTs group included 41 children, 20 boys and 21 girls, mean age (8.2 ± 1.7) years, the age of onset of epilepsy 4.5-11.5 years (the age of onset <8 years in 25 cases, ≥ 8 years in 16 cases). The cognitive characteristics and associated factors were analyzed. The primary data including correct answer numbers and reaction times were analyzed by independent sample t-test between the two groups of children with BECTs and healthy controls based on SPSS 18.0 statistical software.</p><p><b>RESULT</b>Raw data of 9 tasks' scores collected from BECTs and healthy control children were continuous variables in accordance with normal distribution. BECTs children performed significantly worse than controls in choice reaction time ((618+158) vs. (524+254) ms), three-dimensional mental rotation (11 ± 10 vs. 18 ± 12) and visual tracing (10 ± 6 vs.15 ± 6), t=2.01, 3.03 and 3.47, P<0.05, <0.01 and <0.001, respectively.While other 6 tasks showed no significant difference between the two groups (P>0.05 for all comparisons). BECTs boys performed significantly worse than girls on simple substraction tasks compared with standard nine score ((4.7 ± 1.5) vs. (5.6 ± 1.2), t=-2.24, P<0.05). Other 8 tasks showed no significant difference between boys and girls (P>0.05 for all comparisons). Other 9 tasks showed no significant differences between the two groups of BECTs children whose age of onset was before 8 years and those who started seizure ≥ 8 years (P all >0.05). The standard nine scores of simple substraction from the three BECTs groups of dominance sides of spikes and waves during NREM showed significant difference (P<0.05). BECTs children with bilateral discharges performed significantly worse than the other two groups dominantly right or left discharges (4.7 ± 1.2 vs. 6.0 ± 1.2 vs. 4.9 ± 1.4, P all <0.05). There was no significant difference between the two groups with right and left side dominance discharges (P>0.05). Other 8 tasks showed no significant differences among the three groups (P>0.05 for all comparisons).</p><p><b>CONCLUSION</b>Although EEG discharges index below 50% during NREM period, while newly diagnosed BECTs children before treatment with medications against epilepsy performed poorer on tasks of choice reaction time, three-dimensional mental rotation, and visual tracing. The two factors of male and bilateral discharges during NREM period correlate with dysfunction of simple subtraction, the mechanism needs further study and the cognitive function of epilepsy children should be evaluated and followed up, in order to provide psychologic baseline data for persistent cognitive disturbance.</p>

Diagnosis of transposition of the great arteries with posterior aorta by echocardiography / 中华超声影像学杂志

Objective To improve the understanding to transposition of the great arteries with posterior aorta and decrease the clinic misdiagnosis by summarizing their echocardiography features . Methods Echocardiography images were retrospectively reviewed for 7 cases with transposition of the great arteries with posterior aorta who diagnosed in Anzhen Hospital ,and their echocardiography features were summarized .Results In 7 patients ,5 patients had operation ,including 3 for arterial switch operation ,1 for right GLEEN ,1 for right GLEEN and Fortan surgery ,and 2 patients were lost to follow‐up .The echocardiography characters:① Artery short view showed aorta and pulmonary artery existed the half‐surrounding relationship .The aorta was situated posteriorly and the pulmonary artery anteriorly .②Multiple views showed aorta was completely and mostly arised from the right ventricle ,while the pulmonary artery was came off the left ventricle .③ Ventricular septal defects were presented in all cases and aorta could ride the defect of ventricular septal .④ In most cases ,fibrous continuity between the aortic and mitral valve through a ventricular septal defect was found and there was no conus under the aorta .⑤ Some cases could combine the different degree stenosis of the outflow tract of subpulmonary valve and pulmonary valve . Conclusions The connection relationship between two arteries and ventricles can displayed clearly on the long axis view of double outflow tracts of two ventricles under xiphoid process .The acoustic window under the xiphoid process is the key for diagnosis and differential diagnosis of the disease .

Effects of the β-blockers on cardiac protection and hemodynamics in patients with septic shock:a prospective study / 中华危重病急救医学

Objective To investigate the effects of β-blockers on cardiac protection and hemodynamic in patients with septic shock. Methods A prospective randomized controlled trial was conducted. Forty-one patients with septic shock in accordance with early goal directed treatment and met the target within 6 hours,and admitted to intensive care unit (ICU)of Affiliated Huxi Hospital of Jining Medical College from January 2012 to January 2014 were enrolled. The patients were divided into treatment group (n=21)and control group (n=20)by random number table. The patients in both groups were given the standard treatment,esmolol was giving to patients in treatment group in order to control the heart rate (HR)below 100 bpm within 2 hours,and the patients in control group only received standard treatment. The changes in hemodynamic parameters〔mean arterial pressure(MAP),central venous pressure(CVP), HR,cardiac index(CI),stroke volume index(SVI),systemic vascular resistance(SVRI),global end diastolic volume index(GEDVI)〕,biochemistry metabolic of tissue〔central venous oxygen saturation(ScvO2),lactic acid(Lac)〕,and cardiac markers 〔troponin I (cTnI)〕before and 12,24,48,72 hours after the treatment were recorded. Results①Before treatment,the hemodynamic parameters,tissue metabolism index and cTnI had no significant differences in both groups (all P>0.05).②The hemodynamic parameters after treatment in the control group showed no significant difference compared with that before treatment. HR and CI in the treatment group were gradually declined after treatment,SVRI and GEDVI were gradually increased. There were significant differences in HR,CI,SVRI,and GEDVI between treatment group and control group from 12 hours on〔HR(bpm):93±4 vs. 118±13,CI (L·min-1·m-2):3.3 ±0.8 vs. 4.5 ±0.6,SVRI (kPa·s·L-1·m-2):159.2 ±27.4 vs. 130.5 ±24.2,GEDVI(mL/m2):668 ±148 vs. 588 ±103,P0.05). The ScvO2 was not decreased in both groups.④Compared with before treatment,cTnI in the control group was gradually increased,peaked at 72 hours,and that in the treatment group was gradually increased,peaked at 24 hours and then gradually declined. Compared with control group,the cTnI (μg/L)in the treatment group was decreased significantly at 24,48,72 hours (1.15 ±0.57 vs. 1.74 ±0.77,0.93 ±0.52 vs. 2.15 ±1.23,0.52 ±0.36 vs. 2.39 ±1.17,all P<0.01). Conclusionsβ-blockers (esmolol) can improve cardiac function and myocardial compliance,reduce the myocardial injury in patients with sepsis shock. Although β-blockers can decrease cardiac output,it has no influence on the circulation function and tissue perfusion.

Impact of human resistin on glucose uptake in adipocytes and its mechanism / 中华内分泌代谢杂志

The human resistin expression vector was constructed and transfected into cells to observe its effects on glucose uptake in adipocytes,cell proliferation and differentiation.The results suggested that human resistin impaired glucose uptake in adipoeytes via stimulating proliferation of preadipocytes and suppressing adipocyte differentiation.Metformin reversed the inhibition imposed by resistin on adipogenesis.

Clinical Significance of Micrometastasis Detection in Conventional Pathologic Nonmetastasis Lymph Nodes of Patients with Breast Cancer / 医学研究杂志

Objective To study the relationship between lymph node micrometasis and clinicopathological parameters.Methods Immunohistochemical method was adopted to detect cytokeratin pan(CK)、Epithelial Membrane Antigen(EMA) and Carcinoembryonic Antigen(CEA) in 389 lymph nodes from 36 cases of breast cancer, in which conventional pathological diagnosis showed no lymph node metastasis.Results Micrometastasis was found in 38 lymph nodes (9.2) of 12 cases (33.3) .The incidence of micrometasis was correlated with tumor size. Pathological diagonosis and metastasis lymph nodes. Conclusion Micrometastasis detection in negative lymph nodes of breast cancer is recommended to precisely determine the tumor stage, in order to direct cancer therapy and predict prognosis.

Construction of decorin expression vector and its expression in CHO cells / 吉林大学学报(医学版)

Objective To construct a eukaryotic expression vector of decorin(DCN),and observe its expression in CHO cells,in order to provide a basis for further study on the anti-tumor effect of DCN. Methods DCN cDNA was amplified by PCR.The human full-length DCN cDNA ligated into pBluescript was used as template.The fragment was ligated to the expression vector pCDNA3 previously digested with XbaⅠ and EcoR Ⅰ.The ligation mixture was transformed into competent E.coli JM109 cells.Transformants containing inserts were confirmed by restrictive digestion and DNA sequencing.The expression vector was transfected into CHO cells using lipofectamine,and transfected cells were cultivated in DMEM containing G418 (800 mg?L-1) for about 2 months.Immunohistochemistry method was used to detect the expression of DCN protein in stably transfected cells.Results The PCR product was about 1 000 bp.The recombinant expression vector was identified by restrictive digestion and DNA sequencing. DCN protein was detectable in stablely transfected cells.Conclusion The recombinant eukaryotic expression vector pCDNA-DEC is constructed successfully and stablely transfected CHO cells are established.

Influences of recombinant decorin gene transfection on p21~(WAF1/CIP1) of HepG2 hepatoma cells / 吉林大学学报(医学版)

Objective To transfect a recombinant pcDNA3.1-DCN into HepG2 cells and detect the expressions of decorin(DCN) and p21WAF1/CIP1 so as to investigate the mechanism of tumor suppression of DCN.Methods HepG2 cells were divided into pcDNA3.1-dec-HepG2 group(transfected group) and pcDNA3.1-HepG2 group(control group).After the recombinant pcDNA3.1-DCN and pcDNA3.1 were transfected into HepG2 hepatoma cells by liposome,the stably transfected cell lines were established using G418 screening.RT-PCR method was used to detect the expressions of DCN and p21WAF1/ CIP1 mRNA;Western blotting method was used to detect the expressions of DCN and p21WAF1/CIP1 protein;immunohistochemistry was performed to detect the expression of DCN protein.Results Compared with control group,the expressions of DCN and p21WAF1/CIP1 mRNA were significantly increased in transfected group by RT-PCR(P

Expression of ?-galactosidase gene mediated by tetracycline-regulated gene expression system in HepG2 cells / 吉林大学学报(医学版)

Objective To construct the vector for ?-galactosidase (?-gal) gene expression regulation with tetracycline-regulated gene expression system,and to control ?-galactosidase gene expression in hepG2 cells with the vector.So that offer a experimental base for regulating gene expression for liver cancer gene therpy with this system.Methods The primers for explanding two tetracycline operator (TetO2) gene according to the gene nucleotide sequences of TetO2 gene and pcDNA3.1 vector were designed.TetO2 gene was amplified by PCR with pcDNA3.1 plasmid as template.The TetO2 PCR products were cloned into pcDNA3.1 and the vector was named as pcDNA3.1-TetO2.The pcDNA3.1-Teto2 with TetO2 PCR products sequence was analyzed by ABI3130 sequencing analysis.Then ?-gal gene was cloned into pcDNA3.1-TetO2,the vector was named as pcDNA3.1-TetO2-?-gal.The pcDNA6/TR plasmid vector with tetracycline repressor(TR) was transfected into HepG2 cells by Lipotap,and the stable transfection cells were screened by blasticidin.TR gene expression was detected by RT-PCR in HepG-2 cells with and without pcDNA6/TR plasmid transfection.The pcDNA3.1-TetO2-?-gal plasmid vector was transfected into HepG2 cells with and without pcDNA6/TR plasmid transfection,and 3 to 4 d later, these transfected gene cells were treated with doxycline(4 mg?L-1).After 48 h,?-gal gene expression was detected with ?-gal cell staining.Results The pcDNA3.1-TetO2 sequencing analysis results showed that a cassette was made for a cytomegalovirus-type 2 tetracycline operator (TetO2)-TetO2 promoter in pcDNA3.1-Teto2.The double digestion reslut of pcDNA3.1-TetO2-?-gal plasmid vector demonstrated that ?-gal gene was successfully cloned into pcDNA3.1-TetO2 vector .The RT-PCR result of TR gene showed TR gene could be expressed in HepG2 cells with pcDNA6/TR plasmid transfection and TR gene didn’t express in HepG2 cells without pcDNA6/TR plasmid transfection.?-gal gene expressed in HepG2 cells without pcDNA6/TR plasmid transfection,but didn’t express in HepG2 cells with pcDNA6/TR plasmid transfection.However,?-gal gene expression could be induced with doxycline in HepG2 cells with pcDNA6/TR plasmid transfection.Conclusion The tetracycline-regulated gene expression system vector for regulating ?-gal gene expression is successfully constructed and the vector system can control ?-gal gene expression in HepG2 cells.

The perioperative study of neurohormonal factors and hemodynamics in rheumatic valve disease / 中华胸心血管外科杂志

Ovbective To study the perioperative changes in neurohormonal factors and hemodynamics in rheumatic valve disease. Methods Method: The neurohormaonal factors, including catecholamine (CA), angiotesin II (AGII), aldosterone (ALD), atrial natriuretic factor (ANP) and hemodynamics were studies in twenty-four patients with rheumatic valve disease (RVD). 7 congenital heart disease (CHD) as the control group. Results Plasma CA, AGII, ALD and ANP were higher preoperatively in RVD groups than that in CHD group, and significantly rose during cardiopulmonary bypass in RVD groups. Conclusion There is a hyperaldosteronism in patients with mitral valve disease during perioperative period. The high systemic resistance is closely related to significantly high levels of plasma NA and AGII during operation and in the early stage after operation. In patients with mitral valve disease and pulmonary hypertension, most of pulmonary pressure and resistance decrease in the early stage after operation.

Evaluation of proliferative activities in Wilms'tumor / 吉林大学学报(医学版)

Objective: To assess the potential significance of proliferating cell nuclear antigen (PCNA) labelling indexes (PCNA-LI) and AgNORs number in evaluation of proliferative activity of Wilms′ tumor. Methods: A silver staining for AgNORs and an immunohistochemical method PCNA staining were performed on the biopsy specimens taken from 34 children with Wilms′ tumor. Results: No significant differences were observed between PCNA-LI and the pathological types and clinical stages, whereas fraction of S-phase and PI and the number of AgNORs were significantly higher in patients with PCNA-LI≥25% than that in patients with PCNA-LI<25%. The number of AgNORs/cell correlated well with both pathological types and clinical stages. The combination of PCNA-LI and AgNORs can accurately reflect the proliferative activity of cancer cells in Wilms′ tumor. Conclusion: The current pathological types and clinical stages may reflect the aggressive activity in Wilms′ tumor, but insufficient. The simultaneous determination of PCNA-LI and AgNORs count could be used as the essential complementarity of conventional pathological types and stages for accurate evaluation of biologic behaviour of Wilms′ tumor.

Left Heart Geometric Changes and Diastolic Dysfunction in Essential Hypertensive Patients without Left Ventricular Remodeling / 中国医师杂志

Objective To investigate whether the geometry of the left heart and the diastolic function of left ventricle change in patients with essential hypertension but without left ventricular remodeling. Methods Echocardiography was performed in consecutive male patients above 35 years old and without valvular heart disease, myocardial infarction, cardiomyopathy or other diseases that may have effect on the heart structure. The thickness of intra-ventricular septum(IVST) and left ventricular posterior wall (LVPWT), the end-diastolic diameter of the left ventricle(LVEDD), left atrial anterior-posterior(LAAPD), long (LALD) and cross-sectional diameters(LACD) and area(LAA), early diastolic peak velocity(EPV) and atrial contraction peak velocity(APV) of the mitral valve flow were measured. EPV/APV ratio, left ventricular mass index(LVMI), relative wall thickness(RWT) were calculated. Patients with LVMI≥131g/m 2 and/or RWT≥0 44 were excluded from this study. The remaining patients were divided into 2 groups: the patients with essential hypertension were included in the no-remodeling group(group NR), while the patients without essential hypertention entered the normal control group (group NC). Student test was used to compare the variables between the two groups. Results There were 146 cases in group NC and 156 in group NR. IVST,LVPWT,LVEDD,LVMI,LAAPD,LALD,LACD and LAA of group NR were larger than those of group NC(P