Objective To explore the independent risk factors of lymph-node metastasis (LNM) in patients with early gastric cancer (EGC),and establish a risk-prediction model based on LNM.Method 962 early gastric cancer patients undergoing curative radical gastrectomy in the First Hospital of Anhui Medical University from July 2011 to April 2016 were enrolled in this study.The relationships between different clinicopathologic characteristics and LNM were analyzed by Chi-square test or Fisher exact probability,and the independent risk factors were determined using Logistic regression analysis.Moreover,LNM risk was stratified and a risk-predicting model was established on the basis of the identified independent risk factors for LNM.Further,the risk-predicting model was validated using 962 EGC cases.The discriminatory accuracy of risk-predicting model was measured by area under ROC curve (ROC-AUC).Results Mucosal differentiated cancer ≤2 cm,irrespective of the existence of an ulcer,had low LNM rates (LNMR ＜ 3.0％).Univariate and multivariate analysis revealed that female EGC patients with submucosal,undifferentiated,vessel invasion and tumor size ＞ 2 cm were independent risk factors of LNM for EGC patients,and relative risks were 1.893,3.173,1.956,1.922 and 9.027 respectively (P ＜ 0.05).ROCAUC of risk-predicting model was 0.768 (P ＜ 0.01),which showed high diagnostic accuracy and sensitivity.Conclusion Female EGC patients with submucosal undifferentiated carcinomas measuring ＞ 2 cm with vessel invasion have higher risk of LNM.

Human cytochrome P450 (CYP) has a pivotal role on metabolism of xenobiotics and endogenous substances in clinical practice. Since the CYP from human tissue is very complex, and the human tissue itself is not easy to obtain, investigators begin to use all kinds of expression system to heterogeneously express the CYP. The single CYP expressed was then used for drug metabolism and drug-drug interaction research, to improve the efficiency of high-throughput drug screening greatly. Besides, since the polymorphism of drug-metabolizing enzymes makes efficacy variance for some drugs in different population, the heterogeneous expression and drug metabolizing research of certain CYP mutants will be helpful to guide the optimization of therapeutic regimen and conduct the personalized medication.
Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Drug Design ; Drug Evaluation, Preclinical ; Gene Expression ; Humans

OBJECTIVETo investigate the optimal conditions of tri-expression of CYP3A4, POR and cyt b5 in Sf 9 cells.

METHODSThe Sf 9 cells expressing CYP3A4, POR and cyt b5 were cultured in shaker flasks. The optimized conditions, including the temperature and rotation speed, the culture volume, the amount of surfactant and the culture time were studied. The expressed products in microsomes were used to metabolize the testosterone and their metabolic activity was determined.

RESULTSWhen the temperature and rotation speed of the shaker were 27 degree and 90 r/min, the cell density and culture volume were 5X105 cells/ml and 80-120 ml per 250 ml shaker flasks, respectively. When Pluronic F-68 was 0.1% and the culture time was 72 h, the condition was most suitable for culture of Sf 9 cells and expression of targeted proteins. When the ratio of the volume of three added viruses was 1:1:1, the expression condition was optimal, under which the Km, Vmax, and CLint for testosterone metabolism were 119.6 μmol/L,0.52 μmol/(min*g protein) and 4.34 ml/(min*g protein), respectively.

CONCLUSIONThe conditions of tri-expressing of CYP3A4, POR and cyt b5 have been optimized in the study and the product CYP3A4 is obtained with higher metabolic activity.

Animals ; Cytochrome P-450 CYP3A ; biosynthesis ; Cytochromes b5 ; biosynthesis ; Humans ; Insecta ; NADPH-Ferrihemoprotein Reductase ; biosynthesis ; Sf9 Cells

Central South University began to implement the eight-year medical education in 2004, and has accumulated rich experience in the pre-medical education through continuous reform and improvement. Since 2012, Central South University has made a series of reformation, which is more conducive to the all-round development of medical students , on the pre-medical education . Through adding freshman courses, humanities courses, bilingual teaching courses, and applying aca-demic adviser institution, early scientific research training and open courses during the pre-medical education, Central South University has exercised and strengthened students' scientific research ability, the humanistic quality, English level, and made the pre-medical comprehensive examination for the shunt selection of students. Thus Central South University has improved the eight-year medical students' comprehensive quality, and provided an example of the reformation for medical education.

Objective To review the trends of survival rates and complications in extremely low birth weight (ELBW) infants and to improve the prognosis of ELBW infants.Method From January 1999 to December 2015,ELBW infants in our hospital were retrospectively studied.Their survival rates and complications were compared among groups with different birth weight,and the risk factors for survival were identified using multivariate unconditional logistic regression analysis.Result A total of 243 ELBW infant were collected.The median gestational age of ELBW infant was 27.3 weeks (23 ～ 34 weeks),and their median birth weight was 890 g (490 ～ 995 g).Excluding 40 cases refused treatment,the cure and survival rates of the remaining 203 ELBW infants were 43.8％ (89/203) and 65.0％ (132/203),respectively.The survival rate in ELBW infant with birth weight ＜ 600 g was 0/3,increased to 70.8％ (68/96) when birth weight was 900 ～ 999 g,with an ascending trend with increased birth weight (x2 trend =12.673,P ＜0.001).The most common complications of 243 cases were neonatal respiration distress syndrome [87.7％ (213/243)],sepsis [45.3％ (110/243)],intraventricular hemorrhage [37.4％ (91/243)],bronchopulmonary dysplasia [36.6％ (89/243)] and pheumonia [36.6％ (89/243)].The incidence of complications (including intracerebral hemorrhage and hydrocephalus),decreased with increased birth weight.Multivariate unconditional logistic regression analysis found that birth weight below 800 g (＜ 700 g:OR =22.333,95％ CI 1.493 ～ 334.148,P =0.024;700 ～ 799 g:OR =3.573,95％ CI 1.075 ～ 11.874,P =0.038),stage Ⅲ necrotizing enterocolitis (OR =8.803,95％ CI 1.308 ～ 59.244,P =0.025),stage Ⅲ and Ⅳ of intraventricular hemorrhage (OR =8.902,95％ CI 1.127 ～ 70.338,P =0.038) and mechanical ventilation (OR =3.597,95％ CI 1.043 ～ 12.410,P =0.043) were risk factors affecting the ELBW infant's survival.Conclusion As birth weight increases,the survival rate also increases,and the rate of complications decreases.Birth weight,stage Ⅲ necrotizing enterocolitis,stage Ⅲ and Ⅳ intraventricular hemorrhage and mechanical ventilation are risk factors for the ELBW infant's survival.

Objective:To evaluate the clinical feasibility and image quality of three-dimensional breath-hold gradient and spin-echo (3D BH-GRASE) sequence in magnetic resonance cholangiopancreatography (MRCP).Methods:In this prospective study, 59 patients with clinically suspected pancreaticobiliary duct disease performed MRCP with both 3D BH-GRASE and 3D respiration-triggered turbo spin-echo (3D RT-TSE) sequences on 3.0 T scanner in the Affiliated Zhangjiagang Hospital of Soochow University from November 2017 to December 2018. The overall image quality was scored independently by 3 experienced radiologists based on the visibility of different anatomical features of the pancreaticobiliary duct. For comparing the 2 sequences, the statistical difference in scan time was assessed with a paired t test; while subjective scores, signal-to-noise ratios (SNR), contrast ratios (CR) and contrast noise ratios (CNR) were compared with Wilcoxon signed rank test. Results:The scan time of 3D BH-GRASE sequence was 16.4 s while that of 3D RT-TSE was (258.6±42.2) s. Their difference was statistically significant ( t=44.073, P<0.001), with the scan time for 3D BH-GRASE shortened by 94%. The overall quality scores of 3D BH-GRASE images were better than those of 3D RT-TSE ( Z=-6.595, P<0.001). There was no statistical difference ( P>0.05) in the scores regarding the visibility of the upper, middle and lower parts of common bile duct and the first and second branches of left and right hepatic ducts. For visualizing the bottom, body, neck and duct of gallbladder, the 3D BH-GRASE sequence received a higher score than the 3D RT-TSE sequence ( P<0.001). For displaying the proximal, middle and distal segments of main pancreatic duct, the 3D RT-TSE sequence was scored higher than the 3D BH-GRASE sequence ( P<0.05). There was no significant difference of SNR between the two sequences ( Z=0.403, P=0.687), whereas CR and CNR of 3D RT-TSE MRCP were better than those of 3D BH-GRASE MRCP ( Z=6.215, P<0.001 and Z=3.046, P=0.002, respectively). Conclusion:Under the prerequisite of ensuring image quality, a proper use of 3D BH-GRASE sequence can significantly shorten the scan time and thus greatly improve the working efficiency of MRCP examination.

Objective: To study the surgical strategy and clinical efficacy of hypertensive basal ganglia hematomas via transsylvian transinsular approach individually. Methods: The clinical data of 45 patients with hypertensive basal ganglia hematomas underwent microsurgical treatment with different sylvian anatomical points in Jianhu Hospital Affiliated to Nantong University from October 2014 to June 2016 were retrospectively analyzed. Results: The anterior hematomas was dissected through anterior point of lateral fissure, accounted for 66.7%(30 cases), the posterior hematoma was dissected through rolandic points under lateral fissure, accounted for 22.2%(10 cases), the long axis type hematoma was dissected between the anterior point of the lateral fissure and the lower rolandic point, accounted for 11.1%(5 cases). The postoperative CT scan showed that 42 cases were removed the hematomas for more than 90.0%, 3 cases were removed the hematomas for more than 75.0%, there was no postoperative rebleeding.According to GOS score, 14 cases returned to preoperative life status, 20 cases recovered sufficiently to return to family life, 9 cases could ambulate with a crotch but needed assistance, one case showed vegetative survival, one patient died. Conclusion Transsylvian transinsular approach via individual sylvian anatomical point should be advocated to remove basal ganglia hematomas, and it has the advantages of minimally invasion, high hematoma evacuation rate, low rebleeding rate, good neurological recovery and so on.

OBJECTIVETo explore the risk factors of vascular invasion in patients with early gastric cancer (EGC), and to investigate the influence of vascular invasion on the prognosis of EGC patients.

METHODSFrom January 2014 to December 2015, 449 EGC patients underwent curative gastrectomy at the First Affiliated Hospital of Anhui Medical University, of whom 27 cases (6.0%) developed vascular invasion. Clinicopathological and follow-up data of EGC cases were analyzed retrospectively. The association between clinicopathological features and vascular invasion was analyzed by using the Chi-square test or Fisher exact test, and the independent risk factors influencing vascular invasion were identified with logistic regression. The influence of vascular invasion on overall survival was investigated with Kaplan-Meier curve. This study was approved by Ethics Committee of The First Affiliated Hospital of Anhui Medical University (No. 2018-03-12).

RESULTSOf 449 EGC patients, 325 were males and 124 were females (ratio 2.6:1.0) with the mean age of (60.8±10.5) (27 to 87) years; 228 were diagnosed as T1a stage and 221 were diagnosed as T1b. Univariate analysis showed that incidence of vascular invasion in EGC patients with ulceration or scar was 8.4%(18/225), which was higher than 3.8%(9/234) in those without ulceration, and the difference was statistically significant (χ²=4.061, P=0.044). The incidence of vascular invasion in patients with low differentiated tumor was 8.8% (20/226), which was significantly higher than 3.1%(7/223) in those with middle-high differentiated tumor(χ²= 8.363, P=0.012). The incidence of vascular invasion in patients staging T1b was 10.9% (24/221), which was significantly higher than 1.3% (3/228) in those staging T1a (P=0.000); The incidence of vascular invasion in patients with lymph node metastasis was 27.3% (15/55), which was significantly higher than 3.0%(12/394) in those without lymph node metastasis (χ²=50.122, P=0.000). However, there were no significant associations of vascular invasion with gender, age, surgical type, multiple tumor, tumor deposit, tumor location and tumor size (all P > 0.05). Multivariate analysis showed that T1b stage (RR=4.653, 95%CI:1.293-16.747, P=0.019) and lymph node metastasis(RR=7.302, 95%CI: 3.063-17.408, P=0.000) were independent risk factors for vascular invasion in EGC patients. Among 449 EGC patients, 444 received complete follow-up(98.9%), including 26 cases with vascular invasion and 418 cases without vascular invasion. The overall survival in vascular invasion group was significantly lower than that in non-vascular invasion group (χ²=60.463, P=0.000). Besides, 198 EGC patients gained follow-up for 3 years, and the 3-year survival rates of 11 vascular invasion cases and 187 non-vascular invasion cases were 54.5% and 96.8% respectively.

CONCLUSIONSThe risk of vascular invasion is higher in EGC patients with lymph node metastasis and tumor infiltrating the submucosa. The prognosis of EGC patients with vascular invasion is poor.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastrectomy ; Humans ; Lymph Node Excision ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; pathology ; surgery ; Vascular Neoplasms

Objective : To explore the independent risk factors affecting the prognosis，and to construct a nomogram model predicting overall of patients with rectal cancer at T1 and T2 stage． Methods : Retrospective analysis was made on the data of 353 patients diagnosed as rectal cancer，who received the radical rectal resection．The collect- ed data were as follows : age，body mass index (BMI) ，carcinoembryonic antigen ( CEA) ，tumor size，histological type，T stage，N stage，tumor location and number of lymph nodes detected，which were used to perform Kaplan- Meier curve and Log-rank test for univariate analysis and Cox regression for multivariate analysis．The nomogram model was established to predict the overall survival of patients. Results : Age≥60 years，Mucinous adenocarcino- ma，poorly differentiation ，T2 stage ，lymph node metastasis ，BMI ≥25 kg / m2 ，CEA ≥5 μg / L and number of lymph nodes detected ＜12 were associated with overall survival of patients with rectal cancer at T1 and T2 stage (all P＜0. 05) ．Cox regression showed that age≥60 years，T2 stage，mucinous adenocarcinoma，lymph node me- tastasis，CEA≥5 μg / L，BMI ≥25 kg / m2 and lymph node detection number ＜12 were independent risk factors. Based on the above independent risk factors，the nomogram model was constructed，and the predicted curve was in good agreement with the actual survival curve ( C-index = 0. 779) ． Conclusion Age≥60 years，T2 stage，mucin- ous adenocarcinoma，lymph node metastasis，CEA≥5 μg / L，BMI≥25 kg / m2 and the number of lymph nodes de- tected ＜12 are independent risk factors ，and the nomogram established in this study can effectively predict the prognosis of patients with rectal cancer at T1 and T2 stage.

Objective : To elucidate the effect of phosphorylation modification at the threonine 592 (Thr592) site on the inhibition of gastric cancer proliferation by sterile alpha motifs and HD structural domain－containing protein 1 (SAMHD1) and the potential mechanism of action． Methods: Post-translational modifications (PTMs) of SAMHD1 protein in gastric cancer tissues and cell lines in the database were analyzed，and immunohistochemical stai- ning was performed to detect SAMHD1 Thr592 phosphorylation in paired tissues of gastric cancer patients．In gastric cancer cells，SAMHD1 Thr592 variants were constructed and transiently transfected，and cell proliferation was detected using the cell counting kit 8 ( CCK-8 ) method． The phosphorylation of the cyclin-dependent kinases ( CDK) 2 protein threonine 160 (Thr160) site was inhibited by the addition of different concentrations of the CDK6 inhibitor，Palbociclib，which reduced the level of SAMHD1 protein Thr592 phosphorylation．Three online databases were used to analyze the SAMHD1 reciprocal proteins and take the intersection to derive the Nik-related kinase (NRK) protein．Immunoprecipitation ( Co-IP) ，mass spectrometry and Western blot were used to verify the interactions between SAMHD1 and NRK proteins and detect the effect of NRK on the phosphorylation of the SAMHD1 Thr592 site. Results : Compared with PTMs such as ubiquitination，the highest level of phosphorylation modification of SAMHD1 was observed in tumors，and the difference was statistically significant (P＜0. 01) ．Immunohistochemical experiments showed that phosphorylated SAMHD1 (Thr592) was expressed higher in gastric adenocarcinoma than that in normal mucosal tissue adjacent to the cancer，and the difference was statistically significant (P < 0. 01) ．Western blot assay showed that SAMHD1 protein expression was elevated in MKN-45 cells in the overexpression wild type and mutant groups ，and phosphorylated SAMHD1 levels were also elevated in the wild type， T592E and HD / AA groups． CCK-8 assay showed that both SAMHD1 wild type and T592A could inhibit gastric cancer cell proliferation，while T592E and HD / AA had no effect on gastric cancer proliferation． On the basis of overexpression of SAMHD1，CCK-8 suggested that cell proliferation was inhibited after adding different concentrations of Palbociclib treatment，and Western blot assay suggested that the phosphorylation level was also reduced. NRK protein was obtained by Co-IP and mass spectrometry identification to screen the SAMHD1 reciprocal protein profile and database intersection，and NRK was found to interact with SAMHD1 protein and promote phosphorylation at SAMHD1 Thr592 site by Co-IP and Western blot assay． Conclusion Phosphorylation of the Thr592 site contributes to the loss of SAMHD1 's ability to inhibit gastric cancer cell proliferation，which is reversed by Palbociclib．NRK interacts with SAMHD1 protein，promoting phosphorylation of the SAMHD1 Thr592 site.