Application of targeted therapy combined with chemotherapy in advanced gastric cancer / 医学研究生学报

Gastric cancer is one of the most common malignant tumors in the world. Due to its high recurrence rate, gastric cancer is the second leading cause of cancer death. Up to now, surgical treatment is the only method of cure for patients with early disease, and with the development of endoscopy, neoadjuvant radiotherapy and chemotherapy, 5-year survival rate of patients with early gastric cancer could reach more than 95%. However, no specific symptoms were reported in the early stage of gastric cancer, most patients were diagnosed with middle and late stage when they were symptomatic, and no longer candidates for surgical treatment. In recent years, treatment for gastric cancer has evolved rapidly regarding neoadjuvant chemotherapy, molecular targeted therapy and immunotherapy as well as new chemotherapeutic regimens. The representative in the first-line chemotherapy includes platinum and fluorouracil, or a combination of these two agents. Meanwhile, targeted therapy is also widely used in this setting due to its specificity. Thus, it is necessary to summarize the therapeutic effects of current targeted therapy combined with chemotherapy in treating patients with advanced gastric cancer.

Mispronounced consonants and short-term memory among children with developmental speech sound disorder / 中华物理医学与康复杂志

Objective:To observe the pronunciation of consonants among children with developmental speech sound disorder and explore the correlation between mispronounced consonants and short-term memory so as to determine the pathogenesis of the disorder.Methods:Thirty-six children with developmental speech sound disorder and aged 4 to 13 years were evaluated. Their pronunciation of consonants at the phoneme and lexical levels was tested to record the error types and error rate. Twelve of the children were then randomly chosen to form a voice disorder group. Another 10 healthy counterparts constituted a control group. The short-term memory of both groups was assessed and any correlation between pronunciation and short-term memory was analyzed.Results:The children with a developmental speech sound disorder differed significantly from the controls in terms of the numbers of errors in articulating blade-alveolar, blade-palatal and velar consonants. On the phoneme level, the highest substitution error rate occurred when pronouncing lingua-palatal consonants (42.86%), followed by supradental consonants (32%). The highest distortion and non-acquisition error rates were with blade-palatal consonants (14%) and lingua-palatal consonants (9.5%). On the vocabulary level, the highest substitution, distortion, ellipsis and non-acquisition error rates appeared when pronouncing lingua-palatal and velar consonants, velar and blade-palatal consonants, supradental consonants as well as blade-palatal consonants. Significant differences were found between the phoneme and lexical levels in the substitution of supradental and blade-palatal consonants as well as in the ellipsis of blade-alveolar consonants. They were moderately associated with pronunciation level. There was, however, no significant difference in working memory span between the two groups, and no significant correlation was observed between working memory span and pronunciation level.Conclusion:The mispronunciation of consonants by children with developmental speech disorders is higher at the lexical than at the phoneme level. They mainly substitute lingua-palatal and velar consonants and elide supradental consonants, which may be related to short-term memory span.

Possible effect of N-acetyl-leu-leu-norleucinal, an inhibitor of Calpain, on H2O2-induced damage in 661W cells / 中国药理学通报

Aim To investigate the possible effect of N-acetyl-leu-leu-norleucinal (ALLN),an inhibitor of Calpain,on H2O2-induced damage in 661W cells.Methods The cellular survival of 661W treated with different doses of H2O2 without or with ALLN for 24 h was measured by MTT assay.The protein level of Calpain 1 and Calpain 2 was assessed by Western blot.Results Upon the H2O2 treatment at the concentrations of 50,100,500,1 000 μmol · L-1,the survival rate of 661W significantly decreased in a dose-dependent manner compared to that of the control group.Furthermore,the protein level of Calpain 1 and Calpain 2 showed an obviously time-dependent increase in 661W cells treated with 100 μmol · L-1 H2O2 for 12,18,24 h.Finally,the survival rate of 661W treated with ALLN and H2O2 was higher than that treated only with H2O2,and there was no difference in survival rate between ALLN groups (at the concentrations of 25,50,100,200 μmol · L-1) and control group.Conclusions Calpain is involved in the damage induced by H2O2.ALLN,the inhibitor of Calpain,attenuates the oxidative damage,which plays a promising protective role in photo-receptor cells under oxidative stress.

Effects of modified acidic fibroblast growth factor on the proliferation activity cultured hepatocytes in vitro / 安徽医科大学学报

Objective To explore effects of modified acidic fibroblast growth factor (MaFGF) on the growth of hepatocytes cultured in vitro.Methods Hepatocytes of SD rats were isolated,cultured in vitro and subculture for three generations.After being identified,the cells were used in the experiment:① Hepatocytes was observed by inverted phase contrast microscope;② The experiment were randomly divided into control and experimental group,MaFGF with different concentration were added into the culture medium of hepatocytes,after 24,48,72 h,the mitogenic effects were measured by the MTT test;③ The MTT test was used to detect the effects of MaFGF on the proliferation of the hepatocytes in 24 h.The growth curve was plotted.Results ① MaFGF potentiated the proliferation of hepatocytes,which was no significant effect with dose increased.The proliferative rate of 24 h were higher than that of the other time points (P ＜ 0.05).there was statistical difference between the MaFGF group (4.68 × 10-3 ～ 7.80 × 10-3 mg/L) and the control group (P ＜0.05).However,the mitogeoic activities of each MaFGF group were insignificant;② Four days after MaFGF(6.24 × 10-3 mg/L) added into the medium,the population of the hepatocytes was larger than that of the control (P ＜ 0.05).The number of the hepatocytes incubated with MaFGF (12.4 × 104) was 2.1 times as large as the control(6 × 104) in the 10th day(P ＜ 0.05).Conclusion MaFGF has a certain role in promoting the proliferation of the hepatocytes at the appropriate concentration and time.However,the effects don't have concentration deoendent manner.

Expression of ERCC1, MSH2 and PARP1 in non-small cell lung cancer and their prognostic value for patients treated with platinum-based chemotherapy / 肿瘤研究与临床

Objective To analyze the expression of excision repair cross-complementation group 1 (ERCC1),MutS protein homolog 2 (MSH2) and poly ADP-ribose polymerase 1 (PARP1) in non-small cell lung cancer (NSCLC) and their prognostic value for patients receiving platinum-based postoperative adjuvant chemotherapy.Methods Immunohistochemistry was applied to detect the expression of ERCC1,MSH2 and PARP1 in 111 cases of NSCLC paraffin embedded surgical specimens.Through OG-Rank survival analysis,the prognostic value of the ERCC1,MSH2,PARP1 and the related clinic pathological factors were evaluated.Cox regression analysis was used to determine whether ERCC1,MSH2 and PARP1 were independent prognostic factors or not.Results Among the 111 NSCLC patients,the positive expression rates of ERCC1,MSH2 and RARP1 were 33.3 ％(37/111),36.9 ％(41/111) and 55.9 ％(62/111),respectively.Besides,a total of 72 patients who received platinum-based chemotherapy had complete follow-up data.ERCC1 (P＜ 0.001) and PARP1 (P=0.033) were found to be correlated with the survival time while there was no correlation for MSH2 (P =0.298).Patients with both ERCC1 and PARP1 negative had significant longer survival time than those with ERCC1 (P=0.042) or PARP1 (P=0.027) positive alone.Similarly,the survival time of patients with both ERCC1 and PARP1 positive was shorter than those with ERCC1 (P=0.048) or PARP1 (P=0.010) positive alone.Conclusions The NSCLC patients with ERCC1 and (or) PARP1 negative may benefit from platinumbased postoperative adjuvant chemotherapy.The detection of ERCC1 and PARP1 can be used as an important method to assess the prognosis of patients with NSCLC.

Prognostic value of circulating tumor cells and disseminated tumor cells in patients with esophageal cancer: a meta-analysis / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the correlations of circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) with the clinicopathological characteristics, prognostic events, and survival outcomes in esophageal cancer (EC) patients.</p><p><b>METHODS</b>The PubMed, Web of Science, Embase database and Cochrane database were searched for studies reporting the outcomes of interest. The studies were selected according to established inclusion/exclusion criteria. Meta-analysis of the studies was performed using Review Manager 5.3 and Stata12.0 software with the odds ratio (OR), risk ratio (RR) , hazard ratio (HR) , and 95% confidence interval (95% CI) as the effect indexes.</p><p><b>RESULTS</b>Nineteen studies involving a total of 1766 patients were included in the analysis. Significant correlations of CTCs and DTCs were found with the clinicopathological parameters including the tumor stage (OR=1.95), depth of invasion (OR=1.99), lymph node metastasis (OR=2.44), distal metastasis (OR=5.98), histological differentiation (OR=1.67) and lymphovascular invasion (OR=4.48). CTCs and DTCs were also correlated with the prognostic events including relapse (RR=6.86) and metastasis (RR=3.22) and with the survival outcomes including the overall survival (OS) overall analysis (HR=3.46) and disease-free survival/progression-free survival (DFS/PFS) overall analysis (HR=3.00).</p><p><b>CONCLUSION</b>CTCs and DTCs are significantly associated with an advanced tumor stage, depth of tumor invasion, lymph node metastasis, distant metastasis before therapy, differentiation, lymphovascular invasion, relapse and metastasis in patients with EC. They are also significantly correlated with a poorer survival for OS and DFS/PFS to serve as clinical and prognostic predictors in patients with EC.</p>

Expression of YY 1 protein in human insulinoma and its clinical implication / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of Yin Yang 1 (YY1) protein in human insulinoma and explore its clinical significance.</p><p><b>METHODS</b>Nineteen pancreatic neuroendocrine tumor tissue were collected from patients treated in Nanfang Hospital between 2000 and 2014. The protein expression of YY1 in benign and malignant insulinoma tissues were detected by immunohistochemistry.</p><p><b>RESULTS</b>Positive expression for YY1 protein was detected in both benign and malignant tumor tissues, but the malignant tissues had a significantly greater intensity of YY1 expression than the benign tissues (P=0.042). The intensity of YY1 expression was positively correlated with the nature of the tumor, and the insulinomas with high expressions of YY1 had significantly greater malignant potentials (P=0.037).</p><p><b>CONCLUSION</b>The high expression of YY1 protein is associated with the development of insulinima. YY1 may serve as a new tumor marker for detecting the malignant transformation of insulinoma.</p>

Effect of Paidu Baoshen Pill on renal fibrosis in 5/6 nephrectomized rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the anti-renal fibrosis effect of Paidu Baoshen Pill (PBP) on 5/6 nephrectomized rats and to explore its mechanism.</p><p><b>METHODS</b>Totally 50 SD male healthy rats were randomly divided into the normal control group (n = 10), the sham-operation group (n = 10), and the nephrectomy model group (n = 30) according to the proportion of 1:1:3. Rats in the sham-operation group had their renal capsule isolated without nephrectomy. Rats in the nephrectomy model group had their kidneys 5/6 nephrectomized. Then 24 h urine was collected and 24 h urinary protein (24 h UP) detected. Serum blood urea nitrogen (BUN) and serum creatitine (SCr) were also tested. According to the SCr level 30 rats of the model group were further randomly divided into the model group, the PBP group, and the Niaoduqing Granule (NG) group, 10 in each group. Rats in the PBP group and the NG group were respectively administered with PBP (at the daily dose of 1.0 g/kg) and NG (at the daily dose of 3.33 g/kg) by gastrogavage (they were dissolved in distilled water). At the same time, 2 mL distilled water was administered by gastrogavage to rats in the normal control group, the sham-operation group, and the nephrectomy model group, once daily for 4 successive weeks. Mental conditions, activities, hair color, shape of stool, and the body weight were observed during administration. After 4 weeks, urine was collected to detect 24 h UP. Blood was sampled to detect SCr, BUN, transforming growth factor β1 (TGF-β1), type III procollagen (PC III), collagen type IV (Col IV), laminin (LN), and fibronectin (FN). After rats were killed, their left remnant renal tissues were collected for pathological examinations. The protein expression quantity of TGF-β1 and FN was detected by immunohistochemical method. mRNA expression levels of TGF-β1 and FN were detected using real time fluorescent quantitative PCR.</p><p><b>RESULTS</b>There was no statistical difference in the above indices between the normal control group and the sham-operation group (P > 0.05). Compared with the sham-operation group, rats' general condition was poorer in the model group, their body weight grew slower, and 24 h UP increased; serum levels of BUN, SCr, TGF-β1, PC III, Col IV, LN, and FN increased; the residual renal pathological lesion was serious; expression levels of TGF-β1, TGF-β1, mRNA, FN, and FN mRNA increased in the renal tissue (all P < 0.01). Compared with the model group, rats' general condition was better, their body weight grew faster, 24 h UP reduced (P < 0.05), blood levels of BUN and SCr decreased significantly (P < 0.01), serum levels of TGF-β1, PC III, CoL IV, LN, and FN decreased (P < 0.05, P < 0.01); the residual renal pathological lesion was attenuated in the PBP group and the NG group; expression levels of TGF-β1, TGF-β1, mRNA, FN, and FN mRNA decreased (P < 0.01). Compared with the NG group, blood levels of SCr and FN, and expression levels of FN and FN mRNA decreased more in the PBP group (P < 0.05).</p><p><b>CONCLUSIONS</b>PBP had the effect of anti-renal fibro- sis in 5/6 nephrectomized rats. Down-regulating expression levels of TGF-β1, and FN from gene transcription and protein translation levels might be one of its mechanisms.</p>

Structural insights into the enzyme catalysis from comparison of three forms of dissimilatory sulphite reductase from Desulfovibrio gigas.

The crystal structures of two active forms of dissimilatory sulphite reductase (Dsr) from Desulfovibrio gigas, Dsr-I and Dsr-II, are compared at 1.76 and 2.05 Å resolution respectively. The dimeric α2ß2γ2 structure of Dsr-I contains eight [4Fe-4S] clusters, two saddle-shaped sirohaems and two flat sirohydrochlorins. In Dsr-II, the [4Fe-4S] cluster associated with the sirohaem in Dsr-I is replaced by a [3Fe-4S] cluster. Electron paramagnetic resonance (EPR) of the active Dsr-I and Dsr-II confirm the co-factor structures, whereas EPR of a third but inactive form, Dsr-III, suggests that the sirohaem has been demetallated in addition to its associated [4Fe-4S] cluster replaced by a [3Fe-4S] centre. In Dsr-I and Dsr-II, the sirohydrochlorin is located in a putative substrate channel connected to the sirohaem. The γ-subunit C-terminus is inserted into a positively charged channel formed between the α- and ß-subunits, with its conserved terminal Cys104 side-chain covalently linked to the CHA atom of the sirohaem in Dsr-I. In Dsr-II, the thioether bond is broken, and the Cys104 side-chain moves closer to the bound sulphite at the sirohaem pocket. These different forms of Dsr offer structural insights into a mechanism of sulphite reduction that can lead to S3O6(2-), S2O3(2-) and S2-.

Quantification of surface antigens and quantitative flow cytometry.

Measuring expression levels of cell surface antigens is important for the diagnosis of diseases such as B-cell chronic lymphocytic leukemia and the monitoring of targeted therapy, particularly antibody-based therapy. In some cases, the number of antigens that the therapeutic antibodies bind on the cell surface may reflect the efficacy of therapy. Thus, quantitating the number of molecules on the surface of cells before, during, and after therapy would provide important information for monitoring antibody-based therapy and potentially can be used to adjust dosing. We describe a quantitative flow cytometry approach to measuring levels of the CD20 surface antigen, the molecular target of rituximab.

Monitoring cell signaling pathways by quantitative flow cytometry.

As the signaling pathways involved in leukemogenesis are being elucidated, several proteins have emerged as potential targets for therapy. Downstream from those targets are numerous intracellular factors that are constantly modulated. Monitoring those factors could provide insight into the potential efficacy of therapies by predicting which patients will respond to them and by determining the optimal dosage that will inhibit the target protein. We describe a flow cytometry method for quantitation of total and phosphorylated intracellular proteins. Compared with Western blot analysis, this technique dramatically decreases time and labor while providing multiparameter information on specific cell populations. As an example, total and phosphorylated CRKL is quantitated. The methodology has the potential for widespread application in the monitoring of targeted therapy.

Cell-free bead-based detection of total and phosphorylated proteins in plasma and cell lysates: detection of FLT3.

Frequently direct measurement of proteins or their phosphorylation in intact cells is not possible, for instance, when cells are too few, frozen, or subject to degradation. We have demonstrated that tumor cells pour their DNA, RNA, and protein content into circulation because of turnover and breakdown of cell structures. Proteins in solution most likely circulate as complexes, which protects them from degradation. We describe a cell-free, bead-based method that takes advantage of this phenomenon. Our approach is based on immunoprecipitation of the protein of interest on the surface of beads, followed by detection of the protein or its modification (phosphorylation) using a secondary antibody labeled with phycoerythrin at a 1:1 ratio. Fms-like tyrosine kinase-3, which is mutated in majority of cases of acute myeloid leukemia, is used as an example. This method could be applied to the quantitation of several other proteins without the need for intact cells.

Measuring humanized antibodies in plasma of patients treated with antibody-based therapy using bead-based flow cytometry: the story of alemtuzumab.

Alemtuzumab (Campath), the humanized rat monoclonal antibody that targets the CD52 surface antigen, is currently used for treatment of patients with resistant chronic lymphocytic leukemia. Monitoring levels of the antibody in plasma/serum could provide insight into the optimal dosing and scheduling of therapy. Current methods of detecting alemtuzumab in serum or plasma are complicated and difficult to adapt to high-throughput testing. We describe a novel bead-based assay that measures circulating alemtuzumab by taking advantage of remnant rat sequence in the antibody. Levels of total alemtuzumab complexed with CD52, and free alemtuzumab are quantitated in the serum or plasma by flow cytometry. This approach is applicable to the measurement of other humanized antibodies that contain an appropriate remnant animal sequence.

Detection of chromosome translocations by bead-based flow cytometry.

Chromosome translocations resulting in fusion genes have been implicated in leukemogenesis. The paradigm involves the fusion of the genes encoding BCR and ABL, leading to a constitutively active tyrosine kinase. The detection of BCR-ABL has been limited to fluorescence in situ hybridization analysis, reverse transcription-polymerase chain reaction, of mRNA, and Western blot of analysis downstream effectors in the BCR-ABL activated pathway. Here, we describe a novel immunoassay that directly measures levels of BCR-ABL fusion protein and its phosphorylation in peripheral blood plasma and cell lysates. This approach has the potential for widespread application in the detection and quantitation of other fusion genes involved in hematological malignancies.

Clinical observation on effect and safety of combined use of wenxin granule and amiodarone for conversion of auricular fibrillation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the clinical effect of the combined use of amiodarone and Wenxin Granule (WXG) in auricular fibrillation (AF) conversion and its safety.</p><p><b>METHODS</b>Two hundred and twenty-four patients, in whom AF lasted for less than 1 year, were enrolled and randomly assigned into two groups, 112 in each group. Patients in the treated group were treated with WXG and amiodorane and the others in the control group were orally administered with amiodarone alone. The accumulative conversion rate of AF and adverse reaction were monitored during the 6-month observation period.</p><p><b>RESULTS</b>Six-month observation was completed in 109 cases in the treated group and 107 in the control group, while 3 cases and 5 cases in the two groups were dropped out respectively. The difference of accumulative AF conversion rates between the two groups become significant early after one month medication (P < 0.05), and was 65.1% and 47.7% respectively after 6-month of treatment (P < 0.05). Inter-group significant difference was also shown in the aspects of average conversion time, dosage of amiodarone required and the occurrence of adverse reaction (P < 0.05).</p><p><b>CONCLUSION</b>Combined use of WXG and amiodarone has a better effect in improving conversion rate of AF, shortening conversion time and decreasing the required dosage of amiodarone in treating AF as compared with the treatment with amiodarone alone, and by which the adverse reaction of long-term using amiodarone could be avoided.</p>