Objective:To introduce a programmed death-1 (PD-1) inhibitor nivolumab used as a new antitumor agent. Methods:According to the literatures, the action mechanism of nivolumab and the clinical trial results on the main indications approved or being investigated in phase III trials were reviewed and evaluated. Results:Nivolumab could restore the antitumor activity of T cells through binding with PD-1 and consequently blocking its interaction with the key ligands of PD-L1 and PD-L2. Several completed and ongoing clinical trials showed that nivolumab used alone or combined with chemotherapy or CTLA-4 inhibitor ipilimumab exhibited better effica-cy when compared with current clinical used chemotherapy drugs in the treatment of melanoma, non-small cell lung cancer and renal cell carcinoma. Nivolumab was well tolerated during the treatment with such main grade 3-4 adverse events as immune-mediated pneu-monia, abnormal liver functions and fatigue. Conclusion:Through its anti-tumor immune response, nivolumab can improve the clinical efficacy in the treatment of various tumors including melanoma, non-small cell lung cancer and renal cell carcinoma.

Objective To investigate the function and mechanism of embryonic stem cells against Lewis non-small cell lung cancer in vivo. Methods Based on the mouse Lewis non-small cell lung cancer model, we have tested some tumor growth indexes and investigated the immune response of embryonic stem cells against cancer cells. Results Compared with the mice in control group, mice in experimental group received obvious antitumor immunity, which means more activated lymphocytes and antitumor cytokines, resulted in the effective control and inhibition of tumor development. Conclusion Besides the antitumor effect in vitro, embryonic stem cells can also generates immune response in vivo, which could effectively inhibit and/or delay the development of cancer.

Objective Background:MicroRNAs (miRNAs) are naturally occurring small non-coding RNAs,play important roles in cancer initiation and progression.Decreases in miRNAs levels are observed in human cancers,indicating that miRNAs may function intrinsically in tumor suppression.However,the underline mechanisms of miRNA function are little known.Methods MiR-34b in non-small cell lung cancer (NSCLC) tissues was detected using quantitative Real-Time PCR.The relations between miR-34b expression level and clinical pathological parameters were assessed.For in vitro studies,lung cancer cells were transfected with double stranded synthetic miRNA mimics and scrambled controls.Immunohistochemistry technology was explored to validate the related downstream proteins of miR-34b.Results Expression of miR-34b was lower in NSCLC tissues than that in pericarcinous tissues of lung cancer.Additionally,the Spearman correlation test showed lower miR-34b expression was correlated with higher lymph node metastasis (P =0.031).In vitro gain-of-function experiments indicated that miR-34b suppressed cell proliferation by inducing cell apoptosis.IHC results showed relations between lower miR-34b and over-expression of phospho-Met (P =0.012).Conclusion MiR-34b down-regulates Met,following with subsequent changes of downstream p53 and Mdm2,and inversely p53 up-regulates miR-34b in a feedback loop.MiR-34b plays profound roles in progression of NSCLC by inducing apoptosis and decreasing lymph node metastasis.

Objective To explore influence of JAK/STAT3 signaling pathway on the apoptosis of HCC cells.Methods DNA-vector-based RNAi approach silence was used to down-regulate STAT3 expression in Bel-7402 cells.According to the STAT3 cDNA sequence in the GeneBank database,the plasmid pGCsi.U6/neoRFP-STAT3 that was designed for expression of STAT3 siRNA was constructed and synthesized,and then transfected into the Bel-7402 cells with iipofectamine 2000.The apoptotic rate was measured with flow cytometry (FCM) and annexinV/PI apoptosis detection kit staining.The mitochondrial membrane potential (BΨm) was visualized by the JC-1 fluorescence staining and the inverted fluorescence microscope.Moreover,the expression of caspase-3 protein was analyzed by Western blotting.Results The apoptotic ratio of STAT3-siRNA group was (38.82 ± 0.88) ％,which was significantly higher than that in other group [control group (9.22 ± 0.38) ％,scramble-siRNA group (16.47 ± 1.04) ％,P ＜ 0.05].The mitochondrial membrane potential of STAT3-siRNA group observed by the JC-1 fluorescence staining was decreased significantly[(91.33 ± 1.78) ％] and [(89.90 ± 1.92) ％ vs (59.06 ± 1.89) ％,P ＜ 0.05].The Western blot results showed that the protein expression of active caspase-3 in STAT3-siRNA group was significantly higher than other groups (0.48 ± 0.05 vs 0.22 ± 0.04 and 0.26 ± 0.06,P ＜ 0.05).Conclusions STAT3 gene silencing significantly improves the apoptotic effect in the Bel-7402 ceils.

BACKGROUNDLung cancer invading left atrium or base of pulmonary vein belongs to locally advanced lung cancer (T4). The prognosis of treatment without surgery is poor. The aim of this study is to explore the feasibility and the value of surgical method in the treatment of this disease.

METHODSFrom April, 1993 to April, 2005, lobectomy or pneumonectomy combined with extended resection of left atrium were carried out in 46 patients with locally advanced lung cancer. The operations included left low lobectomy in 20 cases, left pneumonectomy in 6 cases, right middle and low lobectomy in 12 cases, right low lobectomy in 3 cases and right pueumonectomy in 5 cases respectively. The base of the pulmonary vein was invaded by the tumor in 34 patients, while left atriums were invaded obviously in 12 patients. Two patients were operated using extracoporeal circulation because of main pulmonary artery and left atrium being invaded. The Kaplan-Meier method (Log rank test) and a COX model were used to analyse the survival and the prognosis.

RESULTSThere was no operative mortality in this series, 15 patients had operative complication, including arrhythmia in 13 cases, pneumonia in 8 cases and heart failure in 1 case. The median survival was 35 months. The 1-, 3-, 5-year survival rates were 84.2%, 43.7%, 30.5% respectively. The survival of patients with N0/1 was better than that of patients with N2 disease, the median survival of them were 38 months and 19 months respectively (P=0.002). Using a Cox model analysis, lymph node stage (N0/1 or N2) was independent prognostic factor, while preoperative chemotherapy, sex, age and the pathologic type were not independent prognostic factors.

CONCLUSIONSSurgical treatment for lung cancer invading the left atrium or the base of pulmonary vein is feasible, especially for N0 patients.

Objective To investigate the correlation between perioperative risk factors including pulmonary fuction indexs and the occurance of postoperative pneumonia in esophageal carcinoma patients,and the prediction efficiency of Peak Expiratory Flow (PEF).Methods Two groups of consecutive esophageal carcinoma patients were included,321 patients in group 1 were devided into postoperative pneumonia group (n =30) and control group (n =291) to screen any relavent risk factom on postoperative pneumonia;group 2 (n =50) was to verify the accurancy and sensitivity of the predictive index.Results he results from group 1 showed that preoperative history of diabetes,previous surgery history,lung function index FEV1 and PEF in the presence of significant differences between the postoperative pneumonia group and the control group,after FDR correction FEV1 and PEF still have statistical significance.Multivariate logistic analysis showed that PEF was an independent prognostic factor of lung infection after esophageal cancer surgery.We build a predictive model with PEF as a variable index of lung infection after esophageal cancer surgery in group 2,the results showed that PEF as a predictor of pulmonary infection has good specificity and sensitivity.Conclusion PEF has a significant correlation with postoperative pulmonary infection in patients with esophageal cancer,and PEF can be used as an effective predictor of postoperative pulmonary infection.

Chaigui granules(CG)are a compound composed of six herbal medicines with significant antidepressant effects.However,the antidepressant mechanism of CG remains unclear.In the present study,we attempted to elucidate the antidepressant mechanism of CG by regulating purine metabolism and purinergic signaling.First,the regulatory effect of CG on purine metabolites in the prefrontal cortex(PFC)of chronic unpredictable mild stress(CUMS)rats was analyzed by ultra high-performance liquid chro-matography tandem mass spectrometry(UHPLC-MS/MS)targeted quantitative analysis.Meanwhile,purinergic receptors(P2X7 receptor(P2X7R),A1 receptor(A1R)and A2A receptor(A2AR))and signaling pathways(nod-like receptor protein 3(NLRP3)inflammasome pathway and cyclic adenosine mono-phosphate(cAMP)-protein kinase A(PKA)pathway)associated with purine metabolism were analyzed by western blotting and enzyme-linked immunosorbent assay(ELISA).Besides,antidepressant mecha-nism of CG by modulating purine metabolites to activate purinergic receptors and related signaling pathways was dissected by exogenous supplementation of purine metabolites and antagonism of puri-nergic receptors in vitro.An in vivo study showed that the decrease in xanthine and the increase in four purine nucleosides were closely related to the antidepressant effects of CG.Additionally,purinergic re-ceptors(P2X7R,A1R and A2AR)and related signaling pathways(NLRP3 inflammasome pathway and cAMP-PKA pathway)were also significantly regulated by CG.The results of exogenous supplementation of purine metabolites and antagonism of purinergic receptors showed that excessive accumulation of xanthine led to activation of the P2X7R-NLRP3 inflammasome pathway,and the reduction of adenosine and inosine inhibited the A1R-cAMP-PKA pathway,which was significantly ameliorated by CG.Overall,CG could promote neuroprotection and ultimately play an antidepressant role by inhibiting the xanthine-P2X7R-NLRP3 inflammasome pathway and activating the adenosine/inosine-A1R-cAMP-PKA pathway.

【Objective】 To investigate the matrix effect on the determination of potency in Recombinant Human Coagulation Factor Ⅷ for Injection (rFⅧ). 【Methods】 Two different detection matrices were used to establish two methods for detecting the potency in Recombinant Human Coagulation Factor Ⅷ for Injection. And the matrix effect on the determination of potency was determined, including specificity, linearity, repeatability, accuracy and intermediate precision. 【Results】 As to the specificity, the recoveries of the two substrates at high vs low concentration level were 112% and 110% vs 104% and 109%, respectively. As to the linearity, in the range of (0.125-1.000) IU/mL, the correlation coefficient between concentration and coagulation time of standard/ sample was higher than 0.99. As to the accuracy/repeatability, the recoveries of two matrices was 104% and 102%, and RSD was 2.4% and 1.9%. As to the intermediate precision, personnel factor of two matrices was 0.72 and 0.23, date factor was 0.79 and 0.85, and RSD(for 12 times) was 4.2% and 3.0%. Comparison of two matrices was as follows: Deviation in test results of 6 batches of rFⅧ was all lower than 5%. There was no significant difference between two matrices. 【Conclusion】 The two matrices for potency detection show good performance including specificity, linearity, repeatability, accuracy, and intermediate precision. They are suitable for the determination of potency in rFⅧ products.

Breast cancer is the most common cancer in women. At present, the in vivo model and traditional cell culture are mainly used in breast cancer researches. However, as high as 90% clinical trials are failed for drugs explored by the above two methods, due to the inherent species differences between humans and animals, as well as the differences in the tissue structure between organs and cells. Therefore, organoid three-dimensional culture is emerging. As a new tumor research model, organoid, a three-dimensional cell complex with spatial structure, has broad application prospects, such as precision medicine, organ transplantation, establishment of refractory disease model, gene therapy and drug research and development. Therefore, organoid is considered as one of the ideal carriers for life science research in the future. Breast cancer, a heterogeneous disease with complex phenotypes, has a low survival rate. Breast cancer organoid can reproduce many key features of human breast cancer, thus, the construction of organoid biological library of breast cancer will provide a new platform for studying the occurrence, development, metastasis and drug resistance mechanism of breast cancer. In this review, we systematically introduce the culture conditions of organoids and their application in breast cancer related research, and the application prospect of organoids.
Animals ; Breast Neoplasms ; Cell Culture Techniques ; Female ; Humans ; Organoids ; Precision Medicine ; Research

OBJECTIVE To study the protective effect of saikosaponin b2(SSb2)on corticosterone(CORT)induced PC12 cell injury and its mechanism.METHODS ① PC12 cells were divided into the cell control group(24 h of culture with RPMI-1640 medium),CORT group(24 h of culture with CORT 100-800 μmol·L-1)and SSb2 group(24 h of culture with SSb2 1.5625,3.125,6.25,12.5,25,50 and 100 μmol·L-1).MTT assay was used to detect the cell survival rate.②PC12 cells were divided into the cell control group(24 h of culture with RPMI 1640 medium),model group(24 h of culture with CORT 400 μmol·L-1),and model+SSb2 group(3 h pretreatment with SSb2 1.5625,3.125,6.25,12.5 and 25 μmol·L-1,removal of the supernatant before cells were co-incubated with CORT 400 μmol·L-1 and corresponding concentrations of SSb2 for 24 h).MTT assay was used to detect the cell survival rate while micro-plate assay was used to detect the lactate dehydrogenase(LDH)leakage rate of PC12 cells.③PC12 cells were divided into the cell control group,model group and model+SSb2 12.5 μmol·L-1 group.AnnexinV-FITC/PI flow cytometry assay was used to detect PC12 cell apoptosis,ultra-perfor-mance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)cell metabonomics was used to detect metabolic profile changes and colorimetric assay was employed to detect the glutamic acid content and glutaminase activity in PC12 cells.RESULTS Compared with the cell control group,the cell viability decreased to(55±6)%(P<0.01)when the concentration of CORT was 400 μmol·L-1.When the concentration of SSb2 was higher than 50 μmol·L-1,there was significant toxicity to PC12 cells(P<0.01).②Compared with the cell control group,the cell survival rate was signif-icantly decreased(P<0.01),while the release rate of LDH was significantly increased(P<0.01)in the model group.Compared with the model group,the cell survival rate significantly increased(P<0.05,P<0.01),while the LDH release rate significantly decreased(P<0.01)in the model+SSb2 group.③ Com-pared with the cell control group,cell apoptosis was significantly increased in the model group(P<0.05).Compared with the model group,cell apoptosis was significantly decreased(P<0.05)in the model+ SSb2 group.Metabolomics results show that SSb2 significantly back-regulated nine differential metabo-lites of glutamate,creatine,N-acetylaspartate,L-tyrosine,citric acid,L-isoleucine,lactic acid,glutamine and choline.Further network analysis of the key metabolites regulated by SSb2 yielded five major metabolic pathways:D-glutamine and D-glutamate metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,alanine,aspartate and glutamate metabolism,tyrosine metabolism and arginine biosynthesis.Compared with the cell control group,the content of glutamate and activity of glutaminase were significantly decreased in the model group(P<0.01).Compared with the model group,the content of glutamate(P<0.01)and activity of glutaminase(P<0.05)were significantly increased in the model+SSb2 group.CONCLUSION SSb2 has a neuroprotective effect on CORT-injured PC12 cells,and the mechanism of which is related to inhibition of apoptosis and regulation of metabolic disorders.