Objective To construct a RhoA-siRNA expression vector and determine its role on the malig-nant behavior of HepG2 cells.Methods A RhoA-siRNA DNA fragment was synthesized and cloned into the expression vector of pGenesil-1.The constructed Rhon-siRNA DNA plasmid was stably transfected into HerG2 cells by lipofectamine,and then HepG2 cells were divided into the HepG2/RhoA-siRNA group (HepG2 cells were transfected with pGenesil-1-RhoA-siRNA),HepG2/control group(HepG2 cells were transfected with control plasmid) and HepG2 group (without plasmid transfection).The inbibitory effect of RhoA-siRNA on RhoA protein expression was shown by Western blot.The proliferation,migration,growth potentiality and cell cycle of transfected HepG2 cells were evaluated by MTT assay,wounded healing,the plate cloning formation test and flow cytometry,respectively.All data were analyzed by one-way analysis of variance (ANOVA) and chi-square test.Results The expression of RhoA protein in the HepG2/RhoA-siRNA group was,significantly decreased compared with that in the other two groups (F=178.19,P<0.05).Scratched cells were healed within 48 hours in the HepG2/control group and HepG2 group,but not in the HepG2/RhoA-siRNA group.The clone formation rates in the HepG2/RhoA-siRNA group,HepG2 group and HepG2/control group were 39%±3%,67%±5%and 70%±6%,respectively,with a significant difference among the three groups(χ2=33.34,38.69,P<0.05).Flow cytometry showed that the number of cells transfected with RhoA-siRNA was highest in the G0/G1 phase and lowest in the S phase(F=70.46,76.57.P<0.05).Conclusion The RhoA-siRNA expression vector can effectively suppress the proliferation and migration of HepG2 cells,which may provide a novel gene therapy for hepatocellular carcinoma.

OBJECTIVETo study on HPLC fingerprint of Propolis and to control it's quality.

METHODThe chromatographic fingerprints of seven samples from different producing areas were determined by RP-HPLC.

RESULTThe chromatograms of Propolis from different producing areas were very similar.

CONCLUSIONThe quality of Propolis can be controlled by determination the HPLC fingerprint.

Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Flavonoids ; analysis ; Materia Medica ; chemistry ; Propolis ; chemistry ; Quality Control ; Reproducibility of Results

BACKGROUNDIntraperitoneal lymphangioma (IL) used to be thought of as a benign lymphatic malformation with a low rate of preoperative diagnosis. This retrospective study aimed to explore the connection between the cysts and clinical manifestation and imaging characteristics, and to study diagnostic confusion, therapeutic principles and potential recurrent reasons, to further enhance the comprehension of this rare disease.

METHODSHere, we retrospectively reviewed 21 patients diagnosed with IL. Age, sex, complaints, physical findings, and imaging features of each patient were documented. The therapies, postoperative complications and treatments were discussed.

RESULTSSymptomatology included eight patients (38%) with intermittent dull pain in the abdomen, and three patients (14%) complained of abdominal persistent pain. The physical examination revealed an abdominal mass in 16 patients (76%), and eight (38%) were reported no discomfort. IL was correctly established preoperatively in 19 patients (90%). Patients were treated using laparotomy, except one who was treated with laparoscopy. Two recurrences were noted during follow-up.

CONCLUSIONSIL should be suspected in any patient with a mobile abdominal mass and surgery is required immediately after discovery of the tumor.

Abdomen ; pathology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Laparoscopy ; Lymphangioma ; diagnosis ; surgery ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies ; Young Adult

OBJECTIVETo study CXCR3 and CCR5 chemokine receptor expression in spleens of patients with primary immune thrombocytopenia (ITP) and its clinical significance.

METHODSThe splenectomy specimens from 10 ITP patients (ITP group) and 8 patients with traumatic splenic rupture (normal control group) were studied. Immunohistochemistry (IHC) was used to study the positive rate of CXCR3 and CCR5. Western blot was performed to detect CXCR3 and CCR5 protein expression, while real-time polymerase chain reaction (RT-PCR) was conducted to analyze their mRNA expression.

RESULTSThe positive rate of CXCR3 and CCR5 were both higher in ITP group (90% and 100%, respectively) than those in control group (75% and 87.5%, respectively)(P < 0.05). The differences were statistically significant (P < 0.05). Protein and mRNA level of CXCR3 in ITP group were 3.0 and 3.5 times as high as those in control group, respectively. Those of CCR5 in ITP group were 1.2 and 1.7 times as high as those in control group, respectively.

CONCLUSIONHigh expression of CXCR3 and CCR5 may play a part in the splenic immune disorders in patients with ITP.

Adolescent ; Adult ; Case-Control Studies ; Child ; Female ; Humans ; Male ; Middle Aged ; Receptors, CCR5 ; metabolism ; Receptors, CXCR3 ; metabolism ; Spleen ; metabolism ; Thrombocytopenia ; immunology ; metabolism ; Young Adult

Objective To identify the clinical characteristics and prognostic factors of young patients with sporadic rectal cancer liver metastasis(RCLM).Methods The clinical data of young RCLM patients at 45 years or under(n=40,as younger patient group)in Peking University First Hospital from January 2016 to January 2021 were reviewed,meanwhile,elder RCLM patient group were comprised of 82 patients older than 45-year-old in a 1:2 ratio.Proportions of categorical variables were compared between young patients and old patients.The clinicopathologic parameters were analyzed with univariate and multivariate Cox regression models and Kaplan-Meier method for demonstrating survival differences between the maximum diameter of liver metastasis and local therapy.Results One hundred and twenty-two RCLM patients were identified,the 1-,3-and 5-year survival rates of young patient group were 97.5%,47.5%,15.0%,those of elder patient group were 84.1%,26.8%,9.8%,respectively.The differences in BMI(P=0.008),primary tumor with obstruction and bleeding(P=0.006),synchronous rectal cancer liver metastases(P=0.005),the maximum diameter of liver metastasis>3 cm(P=0.019)were statistically significant between the two groups.And univariate and multivariate analyses showed that age(P=0.003),N stage(P=0.007),local therapy for liver metastases(P=0.047)and the maximum diameter of liver metastasis(P=0.030)were independent risk factors for influencing the prognosis of RCLM patients;curative resection or not of primary tumor(P=0.035)and the maximum diameter of liver metastasis(P=0.041)were independent risk factors for influencing the prognosis of young RCLM patients.Kaplan-Maier curve demonstrated survival differences between the maximum diameter of liver metastasis and local therapy for liver metastasis in RCLM patients(log-rank P=0.000).Conclusions Although with later staging of initial tumor station,young RCLM patients may obtain better survival benefit compared with old patients.Higher degree of lymph node metastasis,local therapy for liver metastases and the maximum diameter of liver metastasis>3 cm indicates poor prognosis in RCLM patients,and without curative resection of primary tumor and maximum diameter of liver metastasis are also considered as the independent poor prognostic factors of young RCLM patients.Local therapy for liver metastases appears to play an important role in the treatment strategy of RCLM patients.

Objective: Residual SYNTAX score (rSS) can be used as the independent predictor for clinical prognosis and the tool for quantifying incomplete revascularization (IR) in coronary artery disease (CAD) patients with percutaneous coronary intervention (PCI). Our work assessed the prognostic value of rSS on large-scale PCI patients in China. Methods: A total of 10 724 CAD patients undergoing PCI in our hospital in 2013 were studied; 381 patients with previous CABG and hybrid procedure were excluded, 10 343 patients were finally enrolled. Baseline SYNTAX score (bSS) and rSS were calculated before and after PCI. Complete revascularization (CR) was defined by rSS=0 and IR was defined by rSS≥1. The patients were followed-up for 30 months. Clinical endpoint events included MACE, a composite event of all-cause death, myocardial infarction (MI) and revascularization; all-cause death, cardiac death, MI, all-cause death/MI and revascularization.Results: There were 5 050/10 343 (48.8%) patients having CR and 5 293 having IR including 1 908 (18.4%) patients with 1≤rSS≤4, 1 777 (17.2%) with 49. Patients with the higher rSS had more clinical comorbidity and more complicated coronary lesions. Compared with CR patients, IR patients had the higher incidences of 30-month clinical endpoint events. As rSS increasing, the incidence of MACE was elevating accordingly. Multivariate regression analysis indicated that rSS was the independent predictor for MACE and all other endpoints occurrence. Conclusion: IR patients especially those with rSS>9 had the higher incidence of adverse clinical outcomes. rSS has been a good tool for quantifying revascularization and assessing prognosis in PCI patients in China.

OBJECTIVETo investigate the efficacy and advantages of laparoscopy-assisted total gastrectomy (LATG) with D2 dissection of lymph nodes versus conventional open D2 total gastrectomy (OTG) in advanced gastric cancer.

METHODSOne hundred and twenty-five patients with advanced gastric cancer in the middle or upper third of the stomach were operated on from July 2005 to March 2007. Of the patients, 59 cases received LATG and 66 OTG with D2 lymph nodes dissection. Clinical data were recorded and compared between the two groups.

RESULTSNo patient in the LATG group converted to conventional operation with laparotomy. No operation mortality and no severe morbidity occurred in LATG group. As compared with OTG group, in LATG group operation time was longer [(330 +/- 71) min vs. (261 +/- 54) min, P =0.005] in LATG group, but with similar number of lymph node retrieval (36 +/- 13 vs. 34 +/- 16, P =0.450), less operation blood loss [(175 +/- 101) ml vs. (359 +/- 210) ml, P =0.003], earlier recovery of bowel activity (P = 0.015), and a shorter duration of fever after operation (P = 0.024).

CONCLUSIONSLATG with D2 lymph node dissection in advanced gastric cancer is safe and technically feasible with better operative access and visual field, less operation blood loss and earlier recovery.

Adult ; Aged ; Feasibility Studies ; Female ; Gastrectomy ; methods ; Humans ; Laparoscopy ; Lymph Node Excision ; Lymphatic Metastasis ; Male ; Middle Aged ; Stomach Neoplasms ; pathology ; surgery ; Treatment Outcome

OBJECTIVEDiscussing the natural history and the influencing factors of HIV infection among former commercial blood and plasma donors engaged in unsafe blood donation practices in China.

METHODSUsing ambispective cohort study, with data obtained from ten counties (districts) from six provinces in the National AIDS Control Demonstration Area. HIV/AIDS cases were found and confirmed prior to July 24, 2006 being former commercial blood. Plasma donors were selected and data regarding infection, incidence, death, and influencing factors was collected. Analysis was performed using SPSS 12.0 statistical analysis software.

RESULTS(1) In 7551 cases of HIV infection, there were 6533 typical progressors (86.52%, 4757 cases of AIDS), 108 rapid progressors (1.43%), 910 long-term non-progressors (12.05%) with 4865 cases progressed to AIDS (64.43%). The median incubation period for HIV progression to AIDS was nine years (95% CI:8.96-9.04). (2) According to data, from a total of 1157 AIDS cases without ARV therapy (23.78% of total AIDS cases), there were 283 confirmed AIDS-related deaths, of which the median survival time was 6 months (95% CI:4-7) and the two and three year fatality rates were 95% and 99%, respectively. (3) The duration of HIV incubation period was irrespective to gender and age at the time of HIV infection (P > 0.05). Length of survival for untreated AIDS showed correlation to gender (P < 0.05) but no correlation with culture, marital status or age at the time of diagnosis of AIDS (P > 0.05).

CONCLUSIONCompared with the UNAIDS theory regarding slow disease progressors among adults, our study showed a longer AIDS incubation period and shorter outlook for untreated survival, but a similar incubation period for other routes of HIV infection.

Acquired Immunodeficiency Syndrome ; epidemiology ; mortality ; Adolescent ; Adult ; Aged ; Blood Donors ; statistics & numerical data ; China ; Female ; HIV Infections ; epidemiology ; mortality ; Humans ; Infectious Disease Incubation Period ; Male ; Middle Aged ; Young Adult

Objective:To explore the relationship between single nucleotide polymorphism (SNP) rs4977574 in CDKN2B-AS1 gene and female premature coronary artery disease (pCAD) occurrence. Methods: Our research included 2 groups: pCAD group, n=226 consecutive patients≤65 years of age and Control group, n=79 subjects with matched age,without CAD. The genotype of CDKN2B-AS1 SNP rs4977574 was detected by SNaPshot. Blood levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides (TG), uric acid (UA), fasting plasma glucose (FPG) and glycosylated hemoglobin (HbA1c) were examined; relationships between rs4977574 polymorphism and the above parameters were assessed. Results: Compared to Control group, pCAD group had increased blood levels of TG, UA, FPG and HbA1c, P<0.05. With adjusted age, body mass index (BMI), relevant disease history and risk factors, elevated HbA1c (HbA1c>6.2%) obviously increased the risk of female pCAD occurrence (OR=3.35, 95%CI 1.41-8.00, P=0.006). The genotype and allele frequency of rs4977574 were different between pCAD group and Control group, P<0.05. Compared to Control group, pCAD group had the higher frequency of G allele(OR=1.24, 95%CI 1.05-1.48, P=0.019); further analysis found that rs4977574 polymorphism was related to high HbA1c. Compared to AA genotype, GG+GA genotype had the increased incidence of high HbA1c(OR=2.08, 95%CI 1.11-3.89, P=0.022). Conclusion: CDKN2B-AS1 SNP rs4977574 was related to female pCAD occurrence and it was also related to high HbA1c.

Objective To investigate the application prospect of the most extensive genome engineering pig internationally in preclinical xenotransplantation. Methods Porcine endogenous retrovirus (PERV) knockout combined with 3 major heterologous antigen gene knockouts and 9 humanized genes for inhibition of complement activation, regulation of coagulation disorders, anti-inflammatory and anti-phagocytosis were transferred into a pig (PERV-KO/3-KO/9-TG) as a donor, and the heart, liver and kidney were obtained and transplanted to 3 Rhesus macaque recipients respectively to establish a preclinical research model of pig-to-Rhesus macaque xenotransplantation. The functional status of xenografts after blood flow reconstruction was observed and the survival of recipients was summarized. The hemodynamics of xenografts were monitored. The change of hematological indexes of each recipient was compared. The histopathological manifestation of xenografts was observed. Results After the blood flow was reconstructed, all xenografts showed ruddy color, soft texture and good perfusion. The transplant heart, liver and kidney showed full arterial and venous blood flow and good perfusion at 1 d after operation. The postoperative survival time of heart, liver, and kidney transplant recipients was 7, 26, and 1 d, respectively. The levels of creatine kinase, creatine kinase isoenzyme, and lactate dehydrogenase increased in heart transplant recipient at 1 d after operation, and gradually recovered to near normal levels at 6 d after operation. All indexes increased sharply at 7 d after operation. The level of aspartate aminotransferase increased in liver transplant recipients at 2 d after operation, and the alanine aminotransferase basically returned to normal at 10 d after operation, but the total bilirubin continued to increase. Both aspartate aminotransferase and alanine aminotransferase increased at 12 d after operation, and reached a peak at 15 d after operation. The kidney transplant recipient developed mild proteinuria at 1 d after operation, and died of sudden severe arrhythmia. Histopathology showed that the tissue structure of cardiac and renal xenografts was close to normal, and liver xenografts presented with patchy necrosis, the liver tissue structure was disordered, accompanied by inflammatory damage, interstitial hemorrhage and thrombotic microangiopathy. Conclusions PERV-KO/3-KO/9-TG pig shows advantages in overcoming hyperacute rejection, mitigating humoral rejection and coagulation dysregulation. However, whether it can be used as potential donor for clinical xenotransplantation needs further evaluation.