Objective To investigate the value of combined detection of viral capsid antibody(VCA -IgA), early antigen antibody(EA -IgG),and Rta protein antibody IgG(Rta -IgG)in the diagnosis of EB virus associated nasopharyngeal carcinoma(NPC).Methods A total of 47 nasopharynx cancer patients,45 patients with benign rhinitis and 45 healthy controls were recruited.The serum levels of VCA -IgA,EA -IgA and Rta -IgG were tested by enzyme -linked immunosorbent assay (ELISA).Specificity and sensitivity of the indicators alone and combined detection were compared using the clinical diagnosis as the gold standard.Results The expression levels of serum VCA -IgA,EA -IgA and Rta -IgG in the rhinitis group were (0.82 ±0.25),(0.74 ±0.13),(0.89 ±0.27),the levels in the NPC group were (2.16 ±0.39),(1.26 ±0.24),(3.95 ±0.76),and the levels in the healthy control group were (0.65 ±0.14),(0.51 ±0.11),(0.41 ±0.16)respectively.The levels of VCA -IgA,EA -IgA and Rta-IgG in the rhinitis group,NPC group and healthy group were significantly different (F =400.065,232.803, 740.215,P =0.000,0.000,0.000).The levels of VCA -IgA,EA -IgA and Rta -IgG in the NPC patients with different TNMstages were significantly different(F =195.679,30.878,38.561,P =0.000,0.000,0.000),and the trend of each antibody was increased with the severity of the disease.The sensitivity of VCA -IgA was the highest (85.11%),and the specificity of EA -IgA was the highest(95.56%).The sensitivity of the combined assay was 95.74%,which was higher than that of the other three combinations.Conclusion The combination of VCA -IgA, EA -IgA and Rta -IgG can reflect the expression of EBV -associated antigen to a greater extent,and it is superior to single or two combined detection in the diagnosis of EB -associated NPC,with a high clinical value.

Objective To establish an HPLC method for the determination of ginsenosides Rg1 and Re in Shenqi Granula.Methods Chromasil C18 column(250 mm?4.6 mm)was used with acetonitrile-0.05% phosphoric acid solution(21∶79)as mobile phase.The flow rate was 1 mL/min and the detected wavelength was 203 nm.Results Ginsenosides Rg1 and Re could be baseline separated with in 30 min.The average recovery rates were 99.60% and 98.5%,corresponding RSD were 1.93% and 2.31% for ginsenoside Rg1 and Re,respectively(n=5).Conclusion This method is fast and accurate and can be used for quality control of Shenqi Granula.

Objective To construct a lentiviral vector carrying rat sirt1 gene and observe the expression of sirt1 in retinal ganglion cell (RGC) of rat.Methods Rat sirt1 cDNA was inserted into pLV5 vector.After identification by sequencing analysis and PCR,the recombinant sirt1expressinglentivirus vector was packaged by cotransfecting 293T cells with packaged plasmid.Then pLV5-sirt1 was used to infect the cultured Sprague-Dawley rat RGC cell in vitro.The expressions of sirt1 protein and mRNA in infected rat RGC were detected by quantitative real-time PCR and Western blot.Results The sirt1 expression vector pLV5 was successful constructed and sequence was proved to be correct.The expression of sirt1 protein and mRNA in RGC was significantly increased than that in cells infected with control lentiviruses (P ＜ 0.05).Conclusion We have successful constructed a sirt1 expression lentivirus vector pLV5-sirt1 and it can increase the expression of sirt1 protein and mRNA in the rat retinal ganglion cells.

Objective:To investigate the risk factors for lymph node metastasis in patients with early gastric cancer and establish a model for prediction of risk.Methods:The cohort of this retrospective observational study comprised 1096 patients who had undergone radical gastric cancer surgery combined with standard D1 lymphadenectomy and been diagnosed with early gastric cancer by postoperative pathology in Zhongshan Hospital affiliated with Fudan University from January 2016 to July 2022. The patients were allocated to groups with and without lymph node metastases. Clinicopathological characteristics were compared between the two groups and multi-factor logistic regression analysis used to identify independent risk factors for lymph node metastasis in patients with early gastric cancer. Indications for endoscopic resection in the Japanese Gastric Cancer Association (JGCA) guideline were also incorporated into construction of the model. The patient cohort was divided into training and validation sets in a 6:4 ratio. The identified independent risk factors were used to construct a predictive nomogram. Receiver operating characteristic curves were plotted separately and the difference between them in predictive efficacy was compared using the area under the curve (AUC).Results:A total of 1,096 patients with early gastric cancer were included, with 750 males and 346 females. Their average age was (61.4±10.9) years old, and the mean tumor diameter was (23.8±11.4) mm. Among them, 188 patients (17.2%) had positive lymph node metastasis, with 109 cases in N1 stage, 42 cases in N2 stage, and 37 cases in N3 stage. Additionally, 462 patients were in T1a stage, while 634 patients were in T1b stage. Univariate analysis showed that tumor diameter, location, Lauren classification, gross morphology, histological type, intravascular invasion, ulceration, differentiation type and tumor T stage were associated with lymph node metastasis after radical gastrectomy for early gastric cancer (all P<0.05). Multifactorial analysis showed that the presence of intravascular invasion (OR=14.822, 95%CI: 9.323–23.572, P<0.001), undifferentiated type (OR=3.095, 95%CI: 1.649–5.811, P<0.001), tumor T1b (OR=1.798, 95%CI: 1.053–3.079, P=0.032), and tumor diameter ≥2 cm (OR=1.229, 95%CI: 1.031–1.469, P=0.022) were independent risk factors for lymph node metastasis. The baseline data of the training set and validation set were consistent in terms of balance (all P>0.05). We used the above variables to establish a predictive nomogram for lymph node metastasis in patients with early gastric cancer. The AUC values obtained from the validation of the model in the training and validation sets were 0.880 (95%CI: 0.849–0.911) and 0.881 (95%CI: 0.841–0.921), respectively, and were significantly better than the predictive efficacy based on the JGCA guideline (AUC=0.777, 95%CI: 0.746–0.809, P<0.001). Conclusions:Patients with early gastric cancer and intravascular invasion, undifferentiated tumors, tumor T1b, and diameter ≥2 cm are at higher risk of postoperative lymph node metastasis than other patients. The predictive model developed in this study more accurately predicts lymph node metastasis in patients with early gastric cancer than previously proposed methods.

Objective:To investigate the risk factors for lymph node metastasis in patients with early gastric cancer and establish a model for prediction of risk.Methods:The cohort of this retrospective observational study comprised 1096 patients who had undergone radical gastric cancer surgery combined with standard D1 lymphadenectomy and been diagnosed with early gastric cancer by postoperative pathology in Zhongshan Hospital affiliated with Fudan University from January 2016 to July 2022. The patients were allocated to groups with and without lymph node metastases. Clinicopathological characteristics were compared between the two groups and multi-factor logistic regression analysis used to identify independent risk factors for lymph node metastasis in patients with early gastric cancer. Indications for endoscopic resection in the Japanese Gastric Cancer Association (JGCA) guideline were also incorporated into construction of the model. The patient cohort was divided into training and validation sets in a 6:4 ratio. The identified independent risk factors were used to construct a predictive nomogram. Receiver operating characteristic curves were plotted separately and the difference between them in predictive efficacy was compared using the area under the curve (AUC).Results:A total of 1,096 patients with early gastric cancer were included, with 750 males and 346 females. Their average age was (61.4±10.9) years old, and the mean tumor diameter was (23.8±11.4) mm. Among them, 188 patients (17.2%) had positive lymph node metastasis, with 109 cases in N1 stage, 42 cases in N2 stage, and 37 cases in N3 stage. Additionally, 462 patients were in T1a stage, while 634 patients were in T1b stage. Univariate analysis showed that tumor diameter, location, Lauren classification, gross morphology, histological type, intravascular invasion, ulceration, differentiation type and tumor T stage were associated with lymph node metastasis after radical gastrectomy for early gastric cancer (all P<0.05). Multifactorial analysis showed that the presence of intravascular invasion (OR=14.822, 95%CI: 9.323–23.572, P<0.001), undifferentiated type (OR=3.095, 95%CI: 1.649–5.811, P<0.001), tumor T1b (OR=1.798, 95%CI: 1.053–3.079, P=0.032), and tumor diameter ≥2 cm (OR=1.229, 95%CI: 1.031–1.469, P=0.022) were independent risk factors for lymph node metastasis. The baseline data of the training set and validation set were consistent in terms of balance (all P>0.05). We used the above variables to establish a predictive nomogram for lymph node metastasis in patients with early gastric cancer. The AUC values obtained from the validation of the model in the training and validation sets were 0.880 (95%CI: 0.849–0.911) and 0.881 (95%CI: 0.841–0.921), respectively, and were significantly better than the predictive efficacy based on the JGCA guideline (AUC=0.777, 95%CI: 0.746–0.809, P<0.001). Conclusions:Patients with early gastric cancer and intravascular invasion, undifferentiated tumors, tumor T1b, and diameter ≥2 cm are at higher risk of postoperative lymph node metastasis than other patients. The predictive model developed in this study more accurately predicts lymph node metastasis in patients with early gastric cancer than previously proposed methods.

OBJECTIVETo establish a canine cell line with p53 gene knockdown by lentivirus- mediated RNA interference (RNAi).

METHODSFour pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53⁻ silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53⁻) screened using puromycin.

RESULTSThe lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 10⁹ TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53⁻ was established successfully using pGMLV-p53A1 plasmid.

CONCLUSIONThe canine cell line WRD/p53⁻ with stable lentivirus-mediated p53 silencing has been established successfully.

Animals ; Cell Line ; Dogs ; Gene Knockdown Techniques ; Genes, p53 ; Genetic Vectors ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

Objective To explore the value of acoustic radiation force impulse imaging (ARFI) technique in the evaluation of the change of kidney in rabbits with hypothermia and rewarming. Methods Sixteen New Zealand rabbits were placed in -25 ℃ environment for 8 hours, then rewarmed in 23℃ environment. Ultrasonic examinations were performed at room temperature in the time of T0and the time of T1-T8. The right kidney were evaluated by two-dimensional ultrasound and ARFI technique. Results No significant changes were found in the echo and volume of kidney from T1to T8. SWV of kidney descended greatly from T0to T1and raised from T2to T3gradually, then it descended from T3to T8gradually. Significant difference of SWV of kidney were found from T1to T8compared with that at T0(P < 0.05). Conclusions ARFI Technique can estimate the change of kidney in rabbits with hypothermia and rewarming. It can provide more useful information for clinical diagnosis.

Background and objective Increased macrophage inhibitory cytokine-1 (MIC-1), member of trans-forming growth factor-β(TGF-β) superfamily, was found in patients serum with epithelial tumors. hTerefore, our aim was to delineate the diagnostic and prognostic value of serum MIC-1 in patients with stage I-II non-small cell lung cancer (NSCLC). Methods A total of 152 consecutive patients with stage I–II NSCLC were prospectively enrolled and underwent follow up atfer total resection of tumor. Serum MIC-1 level was detected in lung cancer patients by ELISA, 48 benign pulmonary disease patients and 105 healthy controls, and was correlated with clinical features and prognosis of patients. Results hTe level of MIC-1 of NSCLC patients was signiifcantly higher than that of controls (P<0.001) and benign pulmonary disease patients (P<0.001). A threshold of 1,000 pg/mL could be used to diagnose early-stage NSCLC with 70.4%sensitivity and 99.0%speci-ifcity. hTe level of MIC-1 was associated with elder age (P=0.001), female (P=0.03) and T2 (P=0.022). A threshold of 1,465 pg/mL could identify patients with early poor outcome with 72.2%sensitivity and 66.1%speciifcity. hTe overall 3-year survival rate in patients with high level of MIC-1 (≥1,465 pg/mL) was signiifcantly lower than that of patients with low MIC-1 level (77.6%vs 94.8%). Multivariable Cox regression revealed that a high level of MIC-1 was an independent risk factor for compro-mised overall survival (HR=3.37, 95%CI:1.09-10.42, P=0.035). Conclusion High level of serum MIC-1 could be served as a potential biomarker for diagnosis and poorer outcome in patients with early-stage NSCLC.

Objective To establish a canine cell line with p53 gene knockdown by lentivirus-mediated RNA interference (RNAi). Methods Four pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53-silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53-) screened using puromycin. Results The lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 109 TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53- was established successfully using pGMLV- p53A1 plasmid. Conclusion The canine cell line WRD/p53-with stable lentivirus-mediated p53 silencing has been established successfully.

Randomized controlled trials (RCT) have long been considered the gold standard for assessing clinical efficacy. However, RCT are inappropriate for some diseases due to related ethical issues and costs, such as rare diseases that are seriously life-threatening but without adequate treatment. Using real world data (RWD) as external control for RCT could make recruitment less complicated and reduce time and cost. This paper introduces common application scenarios, data sources, study design, basic principles, and statistical methods of RWD as an external control based on the latest guidelines related to RWD and combined with our team's previous research experience. This study could provide references for scholars and sponsors who want to conduct RWD research.