Construction of a nomogram prediction model for survival after radical surgery for intrahepatic cholangiocarcinoma / 中华肝胆外科杂志

Objective:To study the factors influencing survival after radical resection in patients with intrahepatic cholangiocarcinoma (ICC), and to construct a nomogram on survival prediction.Methods:The clinical data of 139 patients with ICC who underwent radical resection at the People's Hospital of Zhengzhou University from June 2018 to December 2021 were retrospectively analyzed. There are 69 males and 70 females, aged (59.5±10.2) years old. These patients were divided into two groups based on a 3: 1 ratio by using the random number method: the test group ( n=104) and the validation group ( n=35). Data from the test group was used to construct a nomagram and data from the validation group was used to validate the predictive power of the nomagram. Univariate and multivariate Cox regression analyses were used to analyse factors influencing survival on the test group patients and to construct a nomogram. The predictive accuracy of the nomogram was determined by receiver operating characteristic (ROC) curves, concordance index (C-index) and calibration curves. Results:The results of the multivariate regression analysis showed that a combined hemoglobin, albumin, lymphocyte and platelet immunoinflammation (HALP) score <37.1 ( HR=1.784, 95% CI: 1.047-3.040), CA19-9 > 35U/ml ( HR=2.352, 95% CI: 1.139-4.857), poorly differentiated tumor ( HR=2.475, 95% CI: 1.237-4.953) and vascular invasion ( HR=1.897, 95% CI: 1.110-3.244) were independent risk factors that affected prognosis of patients with ICC after radical resection (all P<0.05). The AUCs of the nomogram in the test group in predicting the overall survival at 1, 2 and 3 years of patients with ICC after radical resection were 0.808, 0.853 and 0.859, respectively. There was good consistency between the prediction of the nomogram and actual observation. The predicted C-index of the total survival period of the test group was 0.765 (95% CI: 0.704-0.826), and the C-index of the validation group was 0.759 (95% CI: 0.673-0.845). Conclusion:A HALP score <37.1, CA19-9>35 U/ml, poorly differentiated tumour and vascular invasion were independent risk factors for prognosis of ICC patients after radical resection. The nomogram was established based on the above factors and showed good performance in predicting overall survival after radical resection in patients with ICC.

Genome sequencing and analysis of BCG strain for vaccine production in China / 上海预防医学

ObjectiveTo conduct the sequencing and preliminarily analysis of the whole genome of BCG Shanghai D2PB302 strain （hereinafter referred to as BCG Shanghai D2 strain）， which has been used exclusively for the vaccine production in China. MethodsThe DNA of of BCG Shanghai D2 strain （D2-JIA12-1） was extracted， and the whole genome was sequenced by Pacbio-RS Ⅱ. The sequence data was assembled by Smrtlink and polished with the illumina data. Genes， tRNA and rRNA were predicted based on the sequence data. The functional annotation of predicted genes was performed through BLASTP. The IVE-TB antigen gene and MTBVAC were selected as the target sequences to be compared with Mycobacterium tuberculosis H37Rv （NC_000962.3）. ResultsThe sequence length of BCG Shanghai D2 strain was 4 045 232 bp， and the GC content was 65.66%. A total of 4 259 protein-encoding genes were predicted， with an average gene size of 933 bp. 2 476 genes had biological functions and others were hypothetical proteins.144 virulence genes were obtained by comparing with the VFDB. There were 29 type Ⅶ secretion system genes and 10 PE/PPE protein family genes. ConclusionThe whole genome sequence of BCG Shanghai D2 strain is clarified. It lays a broad foundation for subsequent detection of the stability of major antigen genes.

Investigation on the response to Ursodeoxycholic acid in primary biliary cholangitis patients with positive anti-hexokinase 1 antibodies and anti-kelch-like 12 antibodies / 中华风湿病学杂志

Objective:To investigate the value of anti-hexokinase1 antibodies (anti-HK-1) and anti-kelch-like 1 antibodies (anti-KLHL12) antibody in evaluating ursodeoxycholic acid (UDCA) response in patients with primary biliary cholangitis (PBC).Methods:112 PBC patients who had been treated with UDCA for more than 12 months with relatively complete clinical data were analyzed. Serum was collected and the expression of anti-mitochondrial antibody (AMA), anti-HK-1 and anti-KLHL12 antibodies were detected by ELISA. The response to UDCA was based on Paris standard. According to the expression of new antibodies, the patients were divided into the new antibody positive group and negative group. In addition, PBC related baseline indicators were collected, and Spearman correlation analysis was used to study the correlation between antibody expression and baseline indicators in PBC patients.Results:Positivity of anti-HK1 and anti-KLHL12 antibody in AMA-positive PBC patients were 44.7% and 41.2% respectively. Positivity of anti-HK1 and anti-KLHL12 antibodies in AMA negative PBC patients were 33.3% and 22.2%. Anti-HK1 positive patients had higher serum levels of Alaninetransaminase (ALP), aspartate aminotransferase, (AST), γ-glutamyl transpeptidase (γ-GT) and total bilirubin (TBIL) compared with anti-HK1 negative patients, with statistical significant differences ( P<0.05). Notably, correlation analysis showed significantly positive correlation between anti-HK1 antibody expression and ALP, γ-GT and TBIL serum levels ( r=0.735, P<0.05; r=0.332, P<0.05; r=0.491, ( r=0.466, P<0.05). The UDCA response rate in anti-HK-1 antibody positive group was lower than that of the negative group (36.2% vs 60%; P<0.05). Conclusion:Anti-HK-1 and anti-KLHL12 antibody can help to diagnose PBC, and the expression of anti-HK-1 antibody is correlated with the severity of PBC, which could help to predict the reaction of PBC patients to UDCA.

Immune Mechanism of Tumor Cell Lysate Combined with IL-2 Preventing Melanoma and Inhibiting Tumor Growth / 肿瘤防治研究

Objective To investigate the preventive and inhibitory effects of tumor cell lysate(TCL) combined with IL-2 on melanoma and the potential immune mechanism. Methods The B16F10 melanoma TCL cell were prepared using an ultrasonic disruptor. Twenty-four C57BL/6 mice were randomly divided into four groups which were immunized with PBS, IL-2, TCL and TCL+IL-2 for three weeks, and contra lateral tumors were implanted in the fourth week. We observed onset time of tumor and tumor size, collected peripheral blood continuously and monitored the expression of CD4+T and CD8+T cell subsets dynamically by flow cytometry. Spleen and tumor tissues of mice were also tested for CD4+T and CD8+T cell subsets by flow cytometry and immunohistochemistry, respectively. Results The preventive immunization of the TCL+IL-2 group significantly delayed the onset time of tumor (P=0.034); moreover, the tumor volume (P=0.023) and tumor weight (P=0.0015) were also significantly smaller than those in the control group. The expression of CD8+T cell subsets in the TCL+IL-2 group and the TCL-only group were significantly higher than that in the control group (P=0.0016, P=0.012). However, the CD4+T cell subsets of the TCL+IL-2 group decreased after tumor implantation, compared with the control group (P=0.0089). The expression of CD4+T and CD8+T cell subsets in spleen and tumor tissues were as same as those in peripheral blood. Conclusion The tumor vaccine of TCL combined with IL-2 could prevent the occurrence of melanoma in mouse and effectively inhibit tumor growth by activating CD8+T cells.

Expressions of cAMP response element binding protein and phosphorylated cAMP response element binding protein in the hippocampus of drug resistant temporal lobe epileptic rats / 中华神经科杂志

Objective To establish drug resistant models of temporal lobe epilepsy induced by amygdala kindling,and to investigate the changes of cAMP response element binding protein (CREB) and phosphorylated cAMP response element binding protein (p-CREB) expression in the hippocampus tissues in order to explore their roles in drug resistant epileptogenesis.Methods Eighty adult male SD rats were randomly divided into control group (n =10) and model group (n =70).The 70 rats were used to prepare the amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.The successful kindled models were randomly selected as drug resistant epileptic group (n =10) and drug sensitive epileptic group (n =10) according to their response to the phenytoin and phenobarbital.On the basis of behavioral observation,electrophysiology,pathological HE staining,CREB and p-CREB expression changes,we verified the reliability of the models and explored the differences among the three groups above.The changes of CREB and p-CREB expression were detected by immunohistochemical method and Western blotting assay.Results In control group,the electroencephalogram (EEG) frequency was (8.700 ±1.494) Hz;in drug sensitive epileptic group,the EEG frequency was (14.700 ± 1.159) Hz;in drug resistant epileptic group,the EEG frequency was (19.800 ± 1.686) Hz.The frequency differences among the three groups were statistically significant (F =144.202,P =0.000).By immunohistochemical staining,a large number of CREB and p-CREB positive cells were observed in drug resistant epileptic group.As compared with the control group (CREB 0.197 ±0.058,p-CREB 0.260 ±0.176),the expression levels of CREB and p-CREB were increased in drug sensitive epileptic group (CREB 0.361 ±0.151,p-CREB 0.656 ±0.234) and in drug resistant epileptic group (CREB 0.591 ± 0.150,p-CREB 1.077 ± 0.400).The difference among the three groups had statistical significance (F =24.206,20.376,both P ＜ 0.01).Conclusions The expressions of CREB and p-CREB were significantly increased in drug resistant epileptic rats.These findings indicate that the expressions of CREB and p-CREB may play certain roles in the drugresistant epileptogenesis.

Inhibitory effect of juglone on migration and invasion of glioma cell line U-87 MG / 中国神经精神疾病杂志

Objective To investigate the effects and mechanisms of pin1(peptidyl-prolyl isomerase 1) inhibitor ju?glone on migration and invasion in glioma cell line U-87 MG. Methods Glioma cells were treated with juglone at 0, 0.8, 1.6 and 3.2μmol·L-1. Wound-healing assay and invasion assay were performed to examine the inhibitory activity of ju?glone on glioma cell line U-87 MG. Western blot was used to analyze the protein expression of β-catenin, VEGF, MMP-2 and MMP-9. Results The wound-healing assay showed that the wound-healing rate in juglone-treated groups was 46.04%±6.25%and 30.05%±13.35%at concentrations of 1.6μmol·L-1 and 3.2μmol·L-1 , repectively. Juglone treat?ment significantly reduced the wound-healing rate compared with controls (P<0.05). Transwell invasion assay showed that the number of invaded cells in juglone–treated groups was 103.67 ± 5.69 and 77.33 ± 7.77 at the concentrations of 1.6μmol·L-1 and 3.2μmol·L-1 , respectively. Juglone treatment significantly reduced cell invasion compared with con?trols (P<0.05). Treatment with juglone significantly down-regulated the expression levels ofβ-catenin, VEGF, MMP-2 and MMP-9 in U-87 MG cells in a dose-dependent manner. Conclusion The present data suggests that juglone has a significant inhibitory action on cell migration and invasion through down-regulation of theβ-catenin and its downstream VEGF, MMP-2 and MMP-9 protein expressions in glioma cell line U-87 MG.

Inhibitory effect of gossypol on migration of gastric carcinoma cell lines through Akt/β-catenin passway / 中国药理学通报

Aim To investigate the inhibitory effects of gossypol on migration in gastric carcinoma cell lines and its mechanisms. Methods Gastric carcinoma cells were treated with gossypol at different concentra-tions. The effects of gossypol on cells proliferation were measured using the MTT assay. The migration of gas-tric carcinoma cells was detected by transwell assay. The activation of Akt/β-catenin pathway and the ex-pressions of pathway related proteins ( p-Akt,β-cate-nin, cyclin D1, MMP-2, E-cadherin and vimentin ) were detected by Western blot. Results Gossypol treatment could significantly inhibit the proliferation of gastric carcinoma cells in a dose-dependent manner. Transwell assay showed that the migration ability of gastric carcinoma cells was significantly decreased. The inhibitory effect of gossypol on cells migration was more significant than the effect of gossypol on cells prolifera-tion. Compared with the control group, treatment with gossypol significantly suppressed the expressions of p-Akt,β-catenin, cyclin D1, MMP-2 and vimentin pro-tein, whereas the expression of E-cadherin was signifi-cantly up-regulated in gastric carcinoma cells in a dose-dependent manner. Conclusion These results demonstrate that gossypol represses cell migration of gastric carcinoma cells through the down-regulation of the activity of Akt/β-catenin pathway.

Inhibitory effect of tumor growth of recombinant protein fused with cardiac troponin I and artificial peptide / 中国药理学通报

Aim To examine the inhibitory effect of re-combinant cardiac troponin fusion protein composed of subunit I and artificial peptide which was called CIS on tumor growth. Methods The CIS ’ s effect on the growth of human umbilical vein endothelial cells ( HU-VEC) was examined using MTT assay in vitro. Chick chorioallantoic membrane model was applied to study the alteration of angiogenesis treated with purified re-combinant CIS protein. The effect of tumor growth trea-ted with CIS was observed using several in vivo mice xenograft models. Results There was a statistically significant reduction in HUVEC cell proliferative rate when the cells were treated with purified CIS fusion protein, which was also shown in a dose-dependent manner. A decreased amount of new blood vessel for-mation ( angiogenesis) on chick embryo chorioallantoic membranes was observed in recombinant CIS protein treated group compared to the untreated control group. A significant inhibition of tumor growth rate was a-chieved in CIS treated mice compared to CIS untreated control mice in 6 different mouse xenograft models. Conclusions The fusion protein CIS shows the inhibi-tory effect on the tumor growth in our in vivo mouse models, and such inhibition could be mediated by the mechanism of CIS’ s effect on the decrease of HUVEC cell proliferation and further the reduction of angiogen-esis in tumor tissues. This work could provide the foundation for the in-depth investigations on the phar-maceutical application of CIS targeting anti-tumor ther-apy.

Effective of anticancer mouse monoclonal antibody against hMIC-1 targets pancreatic tumor for nude mice in vivo / 中国比较医学杂志

Objective To investigate a antitumor effects of mouse original monoclonal antibody against hMIC -1 as intravenous administration with human pancreatic tumor in vivo and providing experimental data .Methods The fourty-eight mice were randomized into eight groups for loaded with two pancreatic tumor cell lines panc -1 or sw1990 respectively , and individual tumor growth was observed , antitumor efficacy was evaluated after using mouse original monoclonal antibody against hMIC-1 by intravenous administration .The pathological change with formalin fixed , paraffin embedded tissues section was viewed .Results There was a significant difference in tumor volume and weigt in intravenous injection of mouse original monoclonal antibody against hMIC-1 on load pancreatic tumor with nude mice group compared with that in the control group after four week treatment , and the mouse original monoclonal antibody against hMIC-1 demonstrated a close association between inhibition of tumor volume growth and dose-effective in the two xenograft models examined .Under examined microscope , the pancreatic tumor tissue was destroyed evidently in mouse original monoclonal antibody against hMIC-1 group.Conclusions The antitumor effect of intravenous injection for mouse original monoclonal antibody against hMIC-1 is better than that of systemic using gemcitabine .

Research on the impact of compound E-Jiao slurry (CEJS) plus cyclophosphamide in the inhibition rate of lewis pulmonary carcinoma in mice / 国际中医中药杂志

Objective To investigate the effects of Compound E-Jiao Slurry (CEJS) and Cyclophosphamide (CTX) on the inhibition of Lewis pulmonary carcinoma xenografts. Methods Tumor xenografts models were prepared and randomly divided into six groups by weight, including the control group,CTX, CEJS with the dosages of high, middle and low, with a total of ten mice in each group. The control mice were given normal saline 0.4ml by intragastric administration once daily, with a total of twelve times; The mice of CTX group were given CTX 0.2 ml by intraperitoneal injection every other day, with a total of six times; The mice of CEJS group were given CEJS 0.2 ml and saline 0.2 ml by intragastric administration; The mice of combination therapy group 1 were given CEJS 0.4 ml by intragastric administration once daily, and CTX 0.2 ml by intraperitoneal injection every other day; The mice of combination therapy group 2 were given CEJS 0.2 ml by intragastric administration once daily, and CTX 0.2 ml by intraperitoneal injection every other day; The mice of combination therapy group 3 were given CEJS 0.1 ml by intragastric administration once daily, and CTX 0.2 ml by intraperitoneal injection every other day. The weight of tumor xenografts were measured in the experiment,the inhibition rate of tumor xenografts were calculated according to the data after the dissection. Results The combination groups and CTX alone have an inhibition rate of over 50%, and the CEJS group 25.88%. In every experimental group, the weight of tumor showed statistical significance compared with the control (P0.05). Conclusion CEJS can't obviously enhance the sensitivity of tumor xenografts to CTX, but there is a trend for tumor inhibition.

The Construction, Expression and Purification of Therapeutic Adjuvant-free Protein Vaccine——HPV16z-Hsp65-E6/E7 / 中国生物工程杂志

Objective: To develop a therapeutic adjuvantfree protein vaccine against HPV16 which is closely related to cervical cancer of China. Method: First the E6/E7 gene by PCR technology from HPV16z virus strain was isolated in the highrisk cervical cancer area of Shanxi province of China in1990s, and again got the gene segment of Hsp65 from BCG by the same method, mutated the transforming codes in sequences of HPV 16 E6/E7 genes and thus constructed the expression vector pET28aHsp65E6/E7, expressed the Hsp65E6/E7 fusion protein in E.coli BL21(DE3) strain and researched optimal protein purification procedures. Results: The expression vector pET28aHsp65E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification. conclusions: This research laid a foundation for further functional study of the therapeutic adjuvantfree protein vaccine——Hsp65E6/E7.

Evaluation of chemosensitivity of human colorectal cancer determined by APT-TCA assay in vitro / 中华检验医学杂志

Objective To investigate the heterogeneity of chemosensitivity in colorectal cancer using an ATP-tumor chemosensitivity assay (ATP-TCA) and the feasibility of individual chemotherapy.Methods An ATP-TCA were used to determine the effect of 16 single or combined cytotoxic drugs in surgical specimens from 50 patients with colorectal cancer.Results There were considerable differences in chemosensitivity between individuals.The most active single drugs in the assay was identified as Navelbine, Hydroxycamptothecin, 5-Fluorouracil and Paclitaxel; 34.1%, 31.6%, 27.6% and 24.3% of specimens showed sensitivity to them, respectively.5-Fluorouracil+Mitomycin+Aytarabine was found to be the most effective combination, for 100% (11/11)specimens were sensitive to this regimen.5-Fluorouracil+Cisplatin+Adriamycin and Gemcitabine+Cisplatin were moderately active regimens.Conclusions There was the heterogeneity of the in vitro chemosensitivity in colorectal cancer.The use of the ATP-TCA provides a method of selecting appropriate anti-cancer drugs in colorectal cancer.

Effect of Ascitic Fluid Ultrafiltration Refusion and Perfusion of ?-elemene Milk into Abdominal Cavity on Carcinomatous Ascites / 广州中医药大学学报

To investigate the therapeutic effect of ascitic fluid ultrafiltration refusion and perfusion of ?-elemene milk into abdominal cavity on carcinomatous ascites. Seventy-two cases of malignant tumor complicated with carcinomatous ascites were randomized to two groups: group A treated with perfusion of ?-elemene milk into abdominal cavity after ascitic fluid ultrafiltration refusion and group B with perfusion of ?-elemene milk into abdominal cavity after letting-out of ascitic fluid by regular and repeated abdominal paracentesis. Therapeutic effect was compared and ascitic protein (AP) , plasma albumin(PA) content, urine volume and Karnofsky scores were observed before and after treatment as well as the toxic and side effects.In group A, 9 cases were completely relieved (CR) , 22 partially relieved (PR) , 12 not changed (NC) and the effective rate was 86.1%;in group B,4 were CR, 20 PR, 12 NC and the effective rate was 66.7% respectively, the differences being significant (P

Study of therapeutic adjuvant-free protein vaccine-HPV16Z-Hsp65-E6/E7 / 中国肿瘤生物治疗杂志

Objective: To study the biological effects of the HPV16Z-Hsp65-E6/E7 fusion protein vaccine on the tumor associated with HPV16 infection. Methods: We tested the cellular immune responsive intensity to the vaccine by the lymphocyte proliferation and CTL response, and studied the therapeutic effect of the vaccine on mouse TC-1 cell transplanted cancer in vivo and the influence on mouse lifetime. Results: The spleen lymphocytes from the C57BL/6 mouse immunized by the Hsp65-E6/E7 vaccine could proliferate obviously in the presence of the protein and TC-1 tumor cell could be lysed specifically by immune activated lymphocytes in vitro. This animal therapeutic experiment in vivo showed that the vaccine suppressed the growth of TC-1 cell transplanted tumor remarkably. Conclusion: The recombined vaccine can induce specific cellular immune response in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo.