OBJECTIVE:To discuss the effect of trastuzumab combined with chemotherapy on efficacy and serum tumor mark-ers in patients with human epidermal growth factor receptor 2(HER2)positive advanced gastric cancer. METHODS:74 advanced gastric cancer patients in HER2 positive were randomly divided into observation group and control group,37 cases in each group. Control group received cisplatin + capecitabine chemotherapy;based on it,observation group additionally received trastuzumab by intravenous infusion in the first day,8 mg/kg. 21 d was a course,and it lasted for 6 months. The short-term efficacy,long-term effi-cacy,serum tumor marker contents and toxic and side effects were compared between 2 groups. RESULTS:The effective rate (56.76% vs. 32.44%)and control rate(89.19% vs. 65.86%)in observation group were significantly higher than control group,the differences were statistically significant(P<0.05);progression-free survival time and median survival time in observation group sig-nificantly longer than control group,the differences were statistically significant(P<0.05);contents of serum carcinoembryonic anti-gen,carbohydrate antigen (CA)199,CA724 in observation group significantly lower than control group,the differences were statis-tically significant(P<0.05). And there were no significant differences in the incidences of nausea and vomiting,liver damage,bone marrow suppression and hand-foot syndrome in 2 groups (P>0.05). CONCLUSIONS:Trastuzumab combined with chemotherapy helps to reduce serum tumor markers content,and improve short-term efficacy,long-term efficacy of advanced gastric cancer patients with positive HER2.

Objective To detect the expression of KiSS-1, KiSS-1R and MMP-9 in hepatocellular carcinoma (HCC). To study the correlation of KiSS-1, KiSS-1R and MMP-9 expression with invasion and metastasis of HCC, and to explore the underlying mechanisms. Methods The expression of KiSS-1 , KiSS-1R mRNA in 33 HCC samples, 26 non-neoplastic adjacent liver tissue samples and 13 non-neoplastic distant liver tissue samples were detected by RT-PCR. Tissue chips were constructed by modified manual tools, which contained HCC, non-neoplastic adjacent liver tissues, non-neoplastic distant liver tissues, normal liver tissues and intrahepatic metastasis lesions. The expression of KiSS-1 and MMP-9 protein was determined by tissue chips, immunohistochemistry and semi-quantitative image analysis in 150 HCC, 137 non-neoplastic adjacent liver tissue, 98 non-neoplastic distant liver tissues, 16 normal liver tissues and 37 intrahepatic metastasis lesion samples. Results The results of RT-PCR showed that compared with the non-neoplastic adjacent liver tissues and the non-neoplastic distant liver tissues, the expression of KiSS-1 mRNA in HCC was significantly lower (P＜0.01). The expression of KiSS-1R mRNA did not changed in HCC and non-neoplastic liver tissues (P＞0.05). The expression of KiSS-1 protein was lower in HCC with metastasis and in clinical stage Ⅲ than that in those with non-metastasis, and in clinical stages Ⅰ and Ⅱ . It was also higher in the primary than in the metastasis lesions (P＜0.01, respectively). The expression of MMP-9 was higher in tumors having peplos invasion and metastasis than in those with negative peplos invasion and non-metastasis. It was lower in the primary than the metastasis lesions (P＜0. 01, respectively).Negative correlation between KiSS-1 and MMP-9 expression was found in HCC(r=- 0.340,P＜0.01). Conclusions The imbalance between KiSS-1 and MMP-9 expression might play an important role in enhancing the invasive and metastatic capacity of HCC. Loss of KiSS-1 expression might predict an aggressive clinical behavior and was associated with metastatic potential in HCC.

BACKGROUND:Studies have shown that microRNAs (miRNAs) have moderating effect on the renewal and differentiation of cancer stem cels. However, there is no complete understanding on the effect of microRNA-17-92 gene on gastric cancer stem cel renewal and proliferation.
OBJECTIVE:To explore the effect of miRNA-17-92 in promoting self-renewal and proliferation of gastric cancer stem cels.
METHODS:(1) The gradualy reduced miRNAs during gastric cancer stem cel self-renewal were investigated using miRNA array based on RNAs from differentiated and adherent cels. (2) The miRNA-17-92 was constructed and transfected to gastric cancer stem cels. (3) The effects of miRNA-17-92 on the self-renewal of gastric cancer stem cels were studied by tumor sphere assayin vitro. (4) The effects of miRNA-17-92 on the proliferation of gastric cancer stem cels were investigated by MTT assay and colony formation assay.
RESULTS AND CONCLUSION:(1) miR-19b/20a/92a expression gradualy reduced in the adhesion and differentiation of gastric cancer stem cels. (2) The expression of lentivirus carrying miRNA-17-19 gene in MKN28 cels and CD44-/EpCAM- cels were significantly increased; transient transfection of pre-miR-19b/20a/92a increased the expression of CD44-/EpCAM- and MKN28 miRNA, transient transfection of pre-miR-19b/20a/92a antagonists reduced the expression of SGC7901 and CD44+/EpCAM+ miRNA; overexpression of lenti-miR-19b/20a/92a significantly increased the ability of gastric cels to form tumor spheres; chemotherapy drugs prolonged the survival time of lenti-miR-19b/20a/92a-infected cels; transient transfection of pre-miR-19b/20a/92a significantly increased the number of CD44+/EpCAM+ cels, but transfection of pre-miR-19b/20a/92a antagonist reduced the number of CD44+/EpCAM+ cels. (3) MTT proliferation assay showed that gastric cancer cel proliferation rate in miR-19b/20a/92a stably expressing group was faster than that in the control group. Transient transfection of miR-19b/20a/92a precursor accelerated the growth rate of gastric cancer cels, and transient transfection of its antagonist slowed down the growth rate of gastric cancer cels. Colony formation assay showed that transient transfection of miR-19b/20a/92a precursor significantly increased the colony formation number as compared with the control group; transient transfection of miR-19b/20a/92a antagonist reduced the colony formation as compared with the control group. These findings indicate that miR-19b/20a/92a gene presents with continuous deletion in gastric cancer stem cel differentiation process, and miRNA-17-92 gene can promote the renewal and proliferation of gastric cancer stem cels.

Carcinoma, Hepatocellular ; metabolism ; pathology ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; RNA, Messenger ; genetics

OBJECTIVETo detect the expression of survivin protein, survivin mRNA, p27 protein, p27 mRNA and PTEN protein in hepatocellular carcinomas (HCC) and their clinical significances.

METHODSTissue microarrays were constructed. The expression of survivin protein, p27 protein and PTEN protein were evaluated by immunohistochemical methods and in expression of survivin mRNA and p27 mRNA were evaluated by in stiu hybridization respectively in tumor tissues from 141 HCC patients, 128 samples of para-carcinoma liver tissues, 97 liver tissues far from the carcinomas and normal liver tissues from non HCC patients. The relationship of survivin, p27 and PTEN were investigated and a prediction model of HCC was constructed.

RESULTSThe expressions of survivin protein (Ridit 95% CI = 0.689+/-0.048, P < 0.01), survivin mRNA (Ridit 95% CI = 0.690+/-0.049, P < 0.01) and p27 protein (Ridit 95% CI = 0.556+/-0.053, P < 0.05) in HCC tissues were significantly increased, while the expression of PTEN protein (Ridit 95% CI = 0.282+/-0.048) in HCC tissues was significantly reduced (P < 0.01).

CONCLUSIONOverexpressions of survivin mRNA and p27 protein and reduced expression of PTEN protein might be a valuable marker to predict the presence of HCC.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Microtubule-Associated Proteins ; metabolism ; Middle Aged ; PTEN Phosphohydrolase ; metabolism ; RNA, Messenger ; genetics

Objective: To discuss the clinical application and effects of domestic external fixator in the treatment of patients with malformations of limbs. Methods: A total of 7 289 patients with malformation of limbs who had been operated in Qin Sihe orthopedic surgery team from January 1989 to June 2016 were retrospective analyzed. The patients were treated with domestic external fixator, including 4 033 males and 3 256 females, aging from 2 to 82 years with a mean age of 23.4 years. There were 2 732 patients using Ilizarov external fixator, 4 713 patients using hybrid external fixator, 57 patients using monobrachial external fixator, 232 patients using Ilizarov external fixator and hybrid external fixator. The Ilizarov, hybrid and monobrachial external fixator were used in 67, 65 and 0 patients on the upper limbs and in 2 665, 4 616 and 57 patients on the lower limbs. There were 3 028 patients operated on the left limbs, 3 260 patients operated on the right limbs and 1 001 patients operated on the bilateral limbs. The top three types of diseases were sequelae of poliomyelitis, cerebral palsy and post-traumatic stress disorder peromely. Deformity types inclued talipes equinovarus, knee flexion deformity, cavus foot and so on. Results: All the patients were followed up for a period of 2.5 months to 22.4 years, with an average follow-up time of 5.4 years. All of the external fixators were used for single once, and there was no substitute for external fixator quality problem. All the patients were completed surgery goal until removing external fixation except 1 patient gave up treatment and 1 removed the fixator because of metal allergy. The common complications included wire or pin infection and joint movement limitation and so on. Conclusions The domestic external fixator developed and produced based on the characteristics of Chinese limb deformity disability. The domestic external fixator can be used to treat kinds of limb deformities with the advantages of practical, economical, adjustable, universal and portable. The domestic external fixator could meet the clinical demand for fixation of the osteotomy end of the limbs, the correction of the deformity, the repair of the defects and the limb lengthening.

Objective To study the cytotoxicity of bi-chimeric antigen receptors modified T lymphocytes (BiCAR-T) on the human acute myeloid leukemia (AML) cell line HL60 in vitro and the anti-tumor effects of BiCAR-T on the NOD SCID mouse model of AML in vivo.Methods The BiCAR-T were prepared and the expression of chimeric antigen receptor (CAR) of prepared BiCAR-T was analyzed by flow cytometry.In vitro study was divided into two groups:the experiment group (BiCAR-T) and the control group (T lymphocyte).The killing rate of BiCAR-T in vitro on HL60 cells was determined by CCK8 assay and the level of interferon-γ (IFN-γ) secreted from BiCAR-T co-culturing with HL60 cells for 48 hours was detected by enzyme linked immunosorbent assay (ELISA) at different effect/target ratios (5∶1,10 ∶ 1,20 ∶ 1).The NOD SCID mice AML model was established by the injection of HL60 cells through tail vein and used to assess the antitumor effects in vivo.The mice were randomly divided into three groups according to the random number table:the blank control group receiving 0.9％ NaCl 0.2 ml through tail vein,the model group and the treatment group receiving 1 × 107 HL60 cells in 0.2 ml phosphate buffer saline (PBS).After 20 days,the treatment group was injected with 2 × 107BiCAR-T in 0.2 ml PBS 3 times a week for 2 weeks,while the other two groups received 0.9％ NaCl 0.2 ml.The pathological changes in the mice livers and spleens were observed after 2 weeks of treatment.Results The CAR expression rates of BiCAR-T were more than 50.00％.In vitro experiments proved that the killing rates of BiCAR-T in the experimental group and T lymphocytes in the control group on HL60 cells were (25.43 ±1.32)％ vs.(16.18 ±0.75)％,(50.33±3.11)％ vs.(25.47±1.27)％,and (85.89 ± 3.96) ％ vs.(49.45 ± 2.77) ％ at different effect/target ratios (5 ∶ 1,10 ∶ 1,20 ∶ 1).The killing efficiency of BiCAR-T and T lymphocytes on HL60 cells was significantly different (F =404.17,P ＜ 0.001);the killing efficiency of BiCAR-T and T lymphocytes on HL60 cells was significantly different at different effect/ target ratios (F =548.09,P ＜ 0.001);and the killing efficiency on HL60 cells in the experimental group (BiCAR-T) was significantly higher than that in the control group (T lymphocytes) at different effect/target ratios (F =45.36,P ＜ 0.001).The IFN-γlevels secreted from BiCAR-T in the experiment group and T lymphocytes in the control group co-culturing with HL60 ceils after 48 h were (435.65 ± 20.44) pg/ml vs.(356.75 ± 19.87) pg/ml,(1 639.98 ± 95.75) pg/ml vs.(1 109.37 ± 80.98) pg/ml,and (3 467.43 ± 187.54)pg/ml vs.(2 245.52 ± 112.66)pg/ml.The IFN-γlevel in the experiment group (BiCAR-T) and the control group (T lymphocytes) was significantly different (F =156.24,P ＜ 0.001);the IFN-γ level was significantly different at different effect/target ratios (F =857.67,P ＜ 0.001);the IFN-γlevel in the experimental group (BiCAR-T) was significantly higher than that in the control group (T lymphocytes) at different effect/ target ratios of 5 ∶ 1,10 ∶ 1,20 ∶ 1,respectively (F =46.31,P ＜ 0.001).The result of hematoxylineosin staining (HE) staining showed that leukocyte infiltration in the treatment group was significantly decreased compared with the model group.Conclusion The experimental results showed that BiCAR-T is a kind of efficient targeted immunocyte modified by gene engineering,and it can significantly inhibit leukocyte infiltration of AML in vivo and in vitro.