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The present study aimed to investigate the role and underlying mechanisms of long non-coding RNA (lncRNA) muscleblind-like 1 antisense RNA 1 (MBNL1-AS1) in the progression of Prostate cancer (PCa). MBNL1-AS1 and microRNA (miR)-181a-5p expression in PCa tissues and several human PCa cell lines were analyzed, respectively, using StarBasev3.0 project and RT-qPCR assay. After MBNL1-AS1 overexpression, cell proliferation, invasion and migration were, respectively, evaluated using CCK-8, colony formation, transwell and wound healing assays. Dual luciferase assay were used for analysis of the interactions among MBNL1-AS1, miR-181a-5p, and phosphatase and tensin homolog (PTEN). Subsequently, the expression of PTEN and proteins in PI3K/AKT/mTOR signaling was examined using western blot analysis after transfection with miR-181a-5p mimic. The rescue assays were performed to investigate the effects of MBNL1-AS1 and miR-181a-5p on the functions of PCa cells and the expression of PTEN/PI3K/AKT/mTOR signaling by co-transfection with MBNL1-AS1 plasmid and miR-181a-5p mimic. Results indicated that MBNL1-AS1 was conspicuously downregulated while miR-181a-5p upregulating in PCa tissues and cell lines. MBNL1-AS1 overexpression decreased the abilities of cell proliferation, invasion, and migration. Further study revealed that MBNL1-AS1 acted as a sponge for miR-181a-5p and positively regulated PTEN by a sponge effect. Additionally, rescue assays proved that the effect of MBNL1-AS1-upregulation on the proliferation, invasion, and migration of PCa cells was dependent on miR-181a-5p. Furthermore, miR-181a-5p overexpression counteracted the expression of PTEN and proteins in PI3K/AKT/mTOR signaling exerted by MBNL1-AS1-upregulation in PCa cells. This study suggests that MBNL1-AS1 inhibits the progression of PCa via sponging miR-181a-5p and regulating PTEN/PI3K/AKT/mTOR pathway., GRAPHICAL ABSTRACT

Introduction Adjuvant gefitinib therapy prolonged disease-free survival in patients with resected early-stage EGFR-mutation positive NSCLC in the ADJUVANT study (CTONG 1104). However, treatment failure patterns after gefitinib therapy are less well characterized. Methods Overall, 222 stage N1–N2, EGFR-mutant NSCLC patients received gefitinib or vinorelbine plus cisplatin (VP) treatment. Tumor recurrences or metastases occurring during follow-up were defined as treatment failure; sites and data of first treatment failure were recorded. A post hoc analysis of treatment failure patterns which was estimated by Kaplan-Meier and hazard rate curves in modified intention-to-treat patients was conducted. Results There were 114 recurrences and 10 deaths before recurrence across 124 progression events. Spatial distribution analysis showed that the first metastasis site was most frequently the central nervous system in the gefitinib group (29 of 106 [27.4%]), extracranial metastases were most frequent in the VP group (32 of 87 [36.8%]). Temporal distribution analysis showed lower tumor recurrence with gefitinib than with VP 0 to 21 months post-surgery. However, recurrence with gefitinib showed a constant rate of increase 12 months post-surgery. The first peak of extracranial metastasis appeared during 9 to 15 months with VP and 24 to 30 months with gefitinib. The highest peak for central nervous system metastases post-surgery occurred after 12 to 18 months with VP and 24 to 36 months with gefitinib. Conclusions Adjuvant gefitinib showed advantages over VP chemotherapy in treatment failure patterns especially in extracranial metastasis. Adjuvant tyrosine kinas inhibitors may be considered as a treatment option in resected stage N1–N2 EGFR-mutant NSCLC but longer duration should be explored.

The ADJUVANT study reported the comparative superiority of adjuvant gefitinib over chemotherapy in disease-free survival (DFS) of resected EGFR-mutant stage II-IIIA non-small cell lung cancer (NSCLC). However, not all patients experienced favorable clinical outcomes with TKI, raising the necessity for further biomarker assessment. By comprehensive genomic profiling of 171 tumor tissues from the ADJUVANT trial, five predictive biomarkers were identified (TP53 exon4/5 mutations, RB1 alterations, and copy number gains of NKX2-1, CDK4, and MYC. Then we integrated them into the Multiple-gene INdex to Evaluate the Relative benefit of Various Adjuvant therapies (MINERVA) score, which categorized patients into three subgroups with relative disease-free survival and overall survival benefits from either adjuvant gefitinib or chemotherapy (Highly TKI-Preferable, TKI-Preferable, and Chemotherapy-Preferable groups). This study demonstrates that predictive genomic signatures could potentially stratify resected EGFR-mutant NSCLC patients and provide precise guidance towards future personalized adjuvant therapy.

Summary Background Cisplatin-based adjuvant chemotherapy is the standard of care for patients with resected stage II–IIIA non-small-cell lung cancer (NSCLC). RADIANT and SELECT trial data suggest patients with EGFR-mutant stage IB–IIIA resected NSCLC could benefit from adjuvant EGFR tyrosine kinase inhibitor treatment. We aimed to compare the efficacy of adjuvant gefitinib versus vinorelbine plus cisplatin in patients with completely resected EGFR-mutant stage II–IIIA (N1–N2) NSCLC. Methods We did a randomised, open-label, phase 3 trial at 27 centres in China. We enrolled patients aged 18–75 years with completely resected (R0), stage II–IIIA (N1–N2), EGFR-mutant (exon 19 deletion or exon 21 Leu858Arg) NSCLC. Patients were stratified by N stage and EGFR mutation status and randomised (1:1) by Pocock and Simon minimisation with a random element to either gefitinib (250 mg once daily) for 24 months or intravenous vinorelbine (25 mg/m 2 on days 1 and 8) plus intravenous cisplatin (75 mg/m 2 on day 1) every 3 weeks for four cycles. The primary endpoint was disease-free survival in the intention-to-treat population, which comprised all randomised patients; the safety population included all randomised patients who received at least one dose of study medication. Enrolment to the study is closed but survival follow-up is ongoing. The study is registered with ClinicalTrials.gov, number NCT01405079. Findings Between Sept 19, 2011, and April 24, 2014, 483 patients were screened and 222 patients were randomised, 111 to gefitinib and 111 to vinorelbine plus cisplatin. Median follow-up was 36·5 months (IQR 23·8–44·8). Median disease-free survival was significantly longer with gefitinib (28·7 months [95% CI 24·9–32·5]) than with vinorelbine plus cisplatin (18·0 months [13·6–22·3]; hazard ratio [HR] 0·60, 95% CI 0·42–0·87; p=0·0054). In the safety population, the most commonly reported grade 3 or worse adverse events in the gefitinib group (n=106) were raised alanine aminotransferase and asparate aminotransferase (two [2%] patients with each event vs none with vinorelbine plus cisplatin). In the vinorelbine plus cisplatin group (n=87), the most frequently reported grade 3 or worse adverse events were neutropenia (30 [34%] patients vs none with gefitinib), leucopenia (14 [16%] vs none), and vomiting (eight [9%] vs none). Serious adverse events were reported for seven (7%) patients who received gefitinib and 20 (23%) patients who received vinorelbine plus cisplatin. No interstitial lung disease was noted with gefitinib. No deaths were treatment related. Interpretation Adjuvant gefitinib led to significantly longer disease-free survival compared with that for vinorelbine plus cisplatin in patients with completely resected stage II–IIIA (N1–N2) EGFR-mutant NSCLC. Based on the superior disease-free survival, reduced toxicity, and improved quality of life, adjuvant gefitinib could be a potential treatment option compared with adjuvant chemotherapy in these patients. However, the duration of benefit with gefitinib after 24 months might be limited and overall survival data are not yet mature. Funding Guangdong Provincial Key Laboratory of Lung Cancer Translational Medicine; National Health and Family Planning Commission of People's Republic of China; Guangzhou Science and Technology Bureau; AstraZeneca China.

Background: Sophocarpine (SC) is a major alkaloid extracted from Sophora alopecuroides L. Our previous study showed that SC has analgesic effects on neuropathic pain induced by chronic constriction injury (CCI). However, the exact analgesic mechanism of SC on neuropathic pain has not yet been elucidated. Objectives: The current study aimed to examine the anti-neuropathic pain effects of SC on the HMGB1/TLR4/NF-κB signaling pathway to explore its analgesic mechanism in neuropathic pain. Methods: In this experimental study, the neuropathic pain mouse model was established by CCI of the sciatic nerve in a university-affiliated animal lab, China, 2016. Mechanical withdrawal threshold (MWT), thermal withdrawal latency (TWL), cold withdrawal threshold (CWT), and tail-curling latency (TCL) were used to assess the antinociceptive effect of SC in neuropathic pain mice. The mRNA and protein expression levels of HMGB1, TLR4, NF-κB, p-NF-κB, TNF-α, and IL-6 in the spinal cord of neuropathic pain mice were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Results: Treatment with 40 and 20 mg/kg of SC was effective in increasing MWT, lengthening TWL, reducing CWT, and prolonging TCL in mice with neuropathic pain induced by CCI. Compared to the neuropathic-pain model group, treatment with 40 mg/kg of SC could effectively down-regulate HMGB1, TLR4, NF-κB, p-NF-κB, TNF-α, and IL-6 mRNA and protein expression levels in the spinal cord of mice with neuropathic pain induced by CCI. Conclusions: Our results showed that SC has analgesic effects on neuropathic pain induced by CCI, and its analgesic mechanism may be related to down-regulating the HMGB1/TLR4/NF-κB signaling pathway.

The sensitivity of qRT-PCR assay is not adequate for the detection of the samples with lower viral load, particularly in the cerebrospinal fluid (CSF) of patients. Here, we present the development of a highly sensitive real-time nested RT-PCR (RTN RT-PCR) assay in a single closed tube for detection of human enterovirus (HEV). The clinical performance of both RTN RT-PCR and qRT-PCR was also tested and compared using 140 CSF and fecal specimens. The sensitivities of RTN RT-PCR assay for EV71, Coxsackievirus A (CVA)16, CVA6 and CVA10 achieved 10-8 dilution with a corresponding Ct value of 38.20, 36.45, 36.75, and 36.45, respectively, which is equal to traditional two-step nested RT-PCR assay and approximately 2-10-fold lower than that of qRT-PCR assay. The specificity of RTN RT-PCR assay was extensively analyzed insilico and subsequently verified using the reference isolates and clinical samples. Sixteen qRT-PCR-negative samples were detected by RTN RT-PCR and a variety of enterovirus serotypes was identified by sequencing of inner PCR products. We conclude RTN RT-PCR is more sensitive than qRT-PCR for the detection of HEV in clinical samples.

To explore the effect of Norcantharidin on the biological behavior of SW579 cells. The method of MTT is used to detect the inhibition of Norcantharidin with different concentration on the proliferation of SW579 cells. Transwll assay detects the inhibition of cantharidin on the invasiveness of SW579 cells. And ELISA assay detects the related invasion protein MMP2. ELISA method is used to detect VEGF expression level, and to study the inhibition of cantharidin on the angiogenesis. MTT results indicate that cantharidin has inhibition effects on the proliferation of SW579 cell, and with the increase of concentration, the inhibition effects are enhanced. Transwll assay and ELISA assay indicates that cantharidin has inhibition effects on the invasiveness of SW579 cell. The results of VEGF suggest that cantharidin has inhibition effects on angiogenesis.

Background: Skp2 (S-phase kinase-associated protein 2 is overexpressed in many kinds of cancers, and is related to the occurrence and development of tumors. The molecular mechanism of Skp2 in the regulation of bladder cancer cell biological behavior after Skp2 expression knockdown, however, has remained unknown. Objectives: In our present studies (experimental cytobiological studies, we used an RNAi approach to knock down Skp2 expression, and studied its impact on cell proliferation and invasion of T24 cells. Materials and Methods: The expression of the Skp2 gene was knocked down by RNA interference (RNAi) in T24 cells. The transcription level of Skp2 was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis. The expression of Skp2, caspase-3, caspase-8, caspase-9, and p27 (p27Kip1) were measured by western blot assay. Cell proliferation and apoptosis were detected by MTT and flow cytometry. Cell invasion analysis was performed by a matrigel transwell assay. We also detected the level of MMP2 (metalloproteinase-2) and MMP9 (metalloproteinase-9) in cell culture medium by ELISA. Results: The levels of Skp2 mRNA in the negative control group (0.911 ± 0.073) and the blank control group (0.940 ± 0.046) was significantly higher than Skp2 RNAi group (0.185 ± 0.033) (P < 0.001). The levels of Skp2 protein in the negative control group (0.907 ± 0.049) and the blank control group (0.925 ± 0.042) was significantly higher than Skp2 RNAi group (0.220 ± 0.047) (P < 0.001). The proliferation and invasion of T24 cells were significantly inhibited in vitro upon Skp2 RNAi treatment. Conclusions: The proliferation and invasion of human bladder cancer cells can be inhibited by RNAi-targeting Skp2. As a result, Skp2 may be a potential target for gene therapy in cases of human bladder cancer.

Background: Epidemiological studies evaluating the association of two variants rs9340799 and rs2234693 on estrogen receptor 1 (ESR1) with prostate risk have generated inconsistent results. Methods: A meta-analysis was here conducted to systematically evaluate the relationship of these two variants with prostate cancer susceptibility. Results: For rs9340799, heterozygosity of T/C carriers showed a significant increased prostate cancer risk with a pooled odds ratio (OR) of 1.34 (95% CI = 1.06-1.69) while homozygote C/C carriers showed an increased but not statistically significant association with prostate cancer risk (pooled OR = 1.29, 95% CI = 0.94-1.79). Compared to the homozygous TT carriers, the allele C carriers showed a 31% increased risk for prostate cancer (pooled OR = 1.31, 95% CI = 1.06-1.63). No significant association between the rs2234693 and prostate cancer risk was found with the pooled OR of 1.15 (95% CI = 0.97-1.39, T/C and C/C vs. T/T) under the dominant genetic model. Compared to the homozygote T/T carriers, the heterozygous T/C carriers did not show any significantly different risk of prostate cancer (pooled OR = 1.13, 95% CI = 0.94-1.36) and the homozygous C/C carriers also did not show a significant change for prostate cancer risk compared to the wide-type T/T carriers (pooled OR = 1.26, 95% CI = 0.98-1.62). Conclusions: These data suggested that variant rs9340799, but not rs2234693, on ESR1 confers an elevated risk of prostate cancer.

The purpose of this article is to research on whether MACC1 can serve as a potential target for gene therapy of human bladder urothelial carcinoma (BUC). In this study, the expression of MACC1 gene was knocked down by RNA interference (RNAi) in the T24 cell (human BUC cell). The transcription level of MACC1 was detected by RT-PCR. Activities of MACC1, caspase-3, caspase-8, Bax and Met (mesenchymal-epithelial transition factor) protein were measured by Western blot. The cell proliferation and apoptosis were detected by MTT and flow cytometry. The cell’s invasion ability was performed on Matrigel transwell assay. We also detect MMP2 (metalloproteinase-2) proteins by ELISA. The results showed that the level of MACC1 mRNA and protein was significantly reduced after RNAi. MTT assay showed that the proliferation of T24 cell was decreased due to RNA interference. Apoptosis studies also showed that MACC1 gene interference in T24 loses its anti-apoptotic effects. The expression of apoptosis proteins (Caspase-3, Caspase-8 and Bax) increased significantly due to the MACC1 RNAi. The level of Met protein was down-regulated obviously due to RNAi. Transwell assay showed that invasion abilities of T24 cells were reduced obviously due to MACC1 RNAi. Further studies showed that the secretion of MMP-2 was reduced by RNAi. It can conclude that the ability of proliferation and invasion in T24 cells can be inhibited by RNAi-targeting MACC1. As a result, MACC1 can serve as a potential target for gene therapy of human bladder urothelial carcinoma.

Osteopontin (OPN) involves in tumor formation, and strongly correlated with the tumor progression. It was overexpressed in human esophageal squamous cell carcinoma (ESCC). To study the molecular mechanisms of OPN in ESCC, we examined its roles in inhibiting proliferation and invasion of ECA-109 (esophageal squamous cell carcinoma) cells. The expression of OPN gene was knockdown by RNA interference (RNAi) in the Eca-109 cell. The transcription level of OPN was to detect by reverse transcription-quantitative PCR (RT-qPCR). Western blot assay was performed to detect the expression of OPN, Caspase-3,Caspase-8, Caspase-9, ERK1/2, phospho-ERK1/2 and MMP2 after RNAi. The cell proliferation and apoptosis were detected by MTT and Hoechst33342 assay. Transwell inserts was used for detecting ECA-109 cell’s migration ability. The results shown that the level of OPN mRNA and protein was significantly reduced after RNAi. Proliferation and migration of cell line (ECA-109) was significantly inhibited in vitro. The protein phosphorylation and activation of ERK1/2 in the OPN RNAi group reduced significantly than the negative control groups. In Conclusion, the proliferation and migration of human ESCC can be inhibited by RNAi-targeting OPN. OPN can promote the expression of MMP2 through the ERK signaling pathways. OPN could serve as a potential therapeutic target for human ESCC.

Bone degradation is a serious complication of chronic inflammatory diseases such as septic arthritis, osteomyelitis, and infected orthopedic implant failure. Effective therapeutic treatments for bacteria-caused bone destruction are limited. In a previous study, we found that lipopolysaccharide (LPS) induced osteoblast apoptosis and inhibited early and late-stage differentiation of osteoblasts via activation of the C-Jun N-terminal kinase (JNK) pathway. This study aimed to investigate the effect of JNK inhibition by SP600125 on the apoptosis and differentiation of MC3T3-E1 osteoblasts suppressed by LPS. Following pretreatment with SP600125 for 2 h, MC3T3-E1 cells were treated LPS. Following this treatment, cell viability, activity of alkaline phosphatase (ALP) and caspase-3 were measured. mRNA and protein expression of osteoblast-specific genes, mitogen-activated protein kinases (MAPKs), Bax, Bcl-2 and caspase-3 were determined by quantitative polymerase chain reaction (qPCR) and western blot analysis. The results showed that SP600125 significantly restored LPS-inhibited cell metabolism and ALP activity and reduced the upregulated caspase-3 activity of MC3T3-E1 cells induced by LPS. SP600125 also significantly restored the LPS-suppressed mRNA and protein expression levels of early-stage osteoblast-associated genes in a dose-dependent manner. SP600125 significantly downregulated expression of Bax and caspase-3 but upregulated Bcl-2 expression in MC3T3-E1 cells stimulated by LPS. Furthermore, SP600125 selectively triggered the MAPK pathway by reducing the expression of JNK1, while enhancing the expression of extracellular signal-regulated kinase 1 (ERK1). Our results suggested that SP600125 reduced LPS-induced osteoblast apoptosis and restored early-stage differentiation of osteoblasts inhibited by LPS through MAPK signaling. These findings suggest that the therapeutic agent that inhibited JNK1 is of potential use for the restoration of osteoblast function in bacteria-induced bone diseases.

Osteopontin (OPN), a secreted acid glycoprotein with a variety of functions, promotes tumor proliferation, differentiation, invasion and metastasis. The aim of the current study was to investigate whether OPN may serve as a potential therapeutic target for human bladder cancer. RNA interference (RNAi) was performed to downregulate the expression of the OPN gene in T24 human bladder cancer cells. The mRNA and protein expression levels of OPN following RNAi were determined using reverse transcription‑quantitative polymerase chain reaction and western blot analysis, respectively. In addition, the cell cycle progression, apoptosis and proliferation were investigated using by flow cytometric analysis and MTT assay. The cell invasion ability was measured using a Matrigel transwell assay. The mRNA and protein expression levels of OPN were found to be significantly downregulated following RNAi. The proliferation and invasion of T24 cells were significantly inhibited in vitro. In conclusion, RNAi‑targeting OPN may inhibit the proliferation, invasion and tumorigenicity of human bladder cancer cells. Therefore, OPN may serve as a potential therapeutic target for human bladder cancer.

To investigate the genetic characterization of Human parainfluenza virus-3 (HPIV-3) circulating in Gansu and Shaanxi Provinces of China, 719 throat swabs were collected from pediatric patients with acute respiratory infections from 2009-2011. Multiplex RT-PCR was used to screen common respiratory viral pathogens. For HPIV-3-positive specimens, nested RT-PCR was used to amplify the HN gene of HPIV-3. The nucleotides of Hemagglutinin-neuraminidase(HN)gene of 13 HPIV-3 positive strains identified in Gansu and Shaanxi Provinces were successfully sequenced and compared with those downloaded from GenBank. The phylogenetic analysis based on the nucleotides sequence of HN gene showed that 13 HPIV-3 strains belonged to sub-cluster C3 with little sequence variation (overall nucleotide divergence of 0.2%-2.3% and amino acid divergence at 0-1.1%). Compared with the complete gene of HPIV-3 strains from U.S.A., Canada, and Australia, the biggest divergence of the nucleotide and amino acid lovels was 6.0% and 3.4%, respectively. The nucleotide divergence between shaanxi09-2 and shaanxi10-H0091 was 0.9%, while the nucleotide divergence between shaanxi10-H005 and gansull-62110372 was 0.5%, between shaanxi09-2 and BJ/291/09 was 0.6%. However, there was no amino acid divergence among them. It is likely that HPIV-3 virus had been transmitting in Gansu and Shaanxi Provinces for several years. Human parainfluenza virus-3 (HPIV-3) circulated in Gansu and Shaanxi Provinces from 2009 to 2011 belonged to sub-cluster C3.

To understand the evolutionary relationship between the C4a evolutionary lineage of human enterovirus 71 (HEV71) strains circulating in mainland of China during 2008-2010 and 2008 Fuyang strains and study the prevalence and transmission patterns of 2008 Fuyang strains.Download all the complete VP1 (or = 891 bp) or approximate complete VP1 (or = 876 bp) gene nucleotide sequences from GenBank of HEV71 strains circulating in Mainland of China during 2008-2010. And analyze the phylogenetic relationship between Fuyang strains and other provinces' strains using the MEGA software, version 5.0.All of the HEV71 isolates circulating in Mainland of China during 2008-2010 were clustered into evolutionary lineage C4a except for eight strains grouped in the genotype A and one isolate belongs to evolutionary lineage C4b; the homology analysis showed there were 96.5%-100% identity between C4a viruses circulating in mainland China during 2008-2010 and 2008 Fuyang strains, and they were evolved from C4b viruses of 1998. The transmission chains of Fuyang strains were mainly transmitted in Guangdong, Jiangsu, Shanghai, Hunan, Shandong provinces.The predominant viruses circulating in Mainland of China during 2008-2010 were evolutionary lineage C4a of human Enterovirus 71; Fuyang transmission chains mainly distributed in southern of China and the Central China around Anhui provinces.

To investigate the role of Helicobacter pylori (Hp) and its products cytotoxin-associated protein (Cag A), vacuolating cytotoxin (VacA) in childhood acute idiopathic thrombocytopenic purpura (aITP), to evaluate the effect of Hp on their clinical outcome.Subjects were enrolled according to case-control design, including 184 aITP children and 154 healthy controls. They were inquired for demographic characteristics, the risk factors regarding Hp infection and ITP through a uniformed questionnaire. Patients with Hp infection were diagnosed by combined detection of serum Hp antibodies and stool antigens. CagA and VacA proteins were tested by ELISA method. In addition, clinical data and follow-up data of aITP children were collected. Non-conditional logistic regression and t test were applied for statistical analysis.(1) The prevalence of Hp infection in aITP children and controls were 41.30% and 35.71%, respectively. No association between Hp infection and children aITP was found with OR of 1.170 (95%CI: 0.7163 - 1.673) after adjusting for confounding variables. (2) No statistical differences regarding initial platelet counts, megakaryocytes counts and the constituent ratio were found between the aITP children with and without Hp infection (P0.05). (3) No differences regarding initial platelet counts were found between aITP children with and without the expression of CagA (P0.05). The follow-up data showed that 32.88% of aITP children with Hp infection, as well as 29.70% of aITP children without Hp infection developed into cITP. No association between Hp infection and development to cITP was found with adjusted OR 1.171 (95%CI: 0.555 - 2.11 2).The results didn't suggest that Hp is unlikely to play a role in the onset of childhood aITP, and in the development of aITP to cITP.

Background Trastuzumab is currently approved for the clinical treatment of breast and gastric cancer patients with HER-2 positive tumors, but not yet for the treatment of esophageal carcinoma patients, whose tumors typically show 5 ~ 35% HER-2 gene amplification and 0 ~ 56% HER-2 protein expression. This study aimed to investigate the therapeutic efficacy of Trastuzumab in patient-derived esophageal squamous cell carcinoma xenograft (PDECX) mouse models. Methods PDECX models were established by implanting patient esophageal squamous cell carcinoma (ESCC) tissues into immunodeficient (SCID/nude) mice. HER-2 gene copy number (GCN) and protein expression were determined in xenograft tissues and corresponding patient EC samples by FISH and IHC analysis. Trastuzumab anti-tumor efficacy was evaluated within these PDECX models (n = 8 animals/group). Furthermore, hotspot mutations of EGFR, K-ras, B-raf and PIK3CA genes were screened for in the PDECX models and their corresponding patient’s ESCC tissues. Similarity between the PDECX models and their corresponding patient’s ESCC tissue was confirmed by histology, morphology, HER-2 GCN and mutation. Results None of the PDECX models (or their corresponding patient’s ESCC tissues) harbored HER-2 gene amplification. IHC staining showed HER-2 positivity (IHC 2+) in 2 PDECX models and negativity in 3 PDECX models. Significant tumor regression was observed in the Trastuzumab-treated EC044 HER-2 positive model (IHC 2+). A second HER-2 positive (IHC 2+) model, EC039, harbored a known PIK3CA mutation and showed strong activation of the AKT signaling pathway and was insensitive to Trastuzumab treatment, but could be resensitised using a combination of Trastuzumab and AKT inhibitor AZD5363. In summary, we established 5 PDECX mouse models and demonstrated tumor regression in response to Trastuzumab treatment in a HER-2 IHC 2+ model, but resistance in a HER-2 IHC 2+/PIK3CA mutated model. Conclusions This study demonstrates Trastuzumab-induced tumor regressions in HER-2 positive tumors, and highlights PIK3CA mutation as a potential resistance mechanism to Trastuzumab treatment in pre-clinical patient-derived EC xenograft models.

To study the effects of desmopressin (DDAVP) on coagulation factor Ⅷ (FⅧ) and activated partial thromboplastin time (APTT) in children with mild hemophilia A.Eighteen children with mild hemophilia A were enrolled. DDAVP (0.3 μg/kg•d) was injected intravenously for 5 days. Plasma FⅧ levels and APTT were measured before and after DDAVP treatment.In 16 of 18 children with mild hemophilia A, the bleeding symptoms, including the articular or musclar hematoma, were significantly alleviated as a result of DDAVP treatment. The plasma FⅧ levels increased significantly to (27±4)% and APTT was shortened to (66±10)s 60 minutes after the first dose of DDAVP treatment. The plasma FⅧ remained at the levels of 25%-30% during 3-4 days of DDAVP treatment. Five days after DDAVP treatment, the plasma FⅧ levels decreased [(21±3)%], and APTT was prolonged when compared with 1-4 days of DDAVP treatment.DDAVP treatment can increase plasma FⅧ levels and shorten APTT in children with mild hemophilia A. DDAVP is effective in the treatment of mild hemophilia A. The duration of DDAVP therapy for mild hemophilia A is recommended as 3 to 4 days.

We want to explore the harm degree of human rhinovirus in infants in Beijing area.From May 2008 to September 2009, 240 nasopharyngeal aspirates were collected from the children and infants who were hospitalized and with lower respiratory tract infections. These specimens were screened for HRV by real-time reverse transcription PCR (RT-PCR) and statistically analysised.In all of 240 hospitalized children, 208 cases were admission diagnosis of pneumonia, accounting for 86.67% (208/240), no deaths, the ratio of male and female patients was 1.93 : 1, and the collected samples reached to a maximum number in February 2009. Real-time PCR used to detect human rhinovirus, positive samples number is 71, positive rate is 29.58% (71/240), and the main symptoms and clinical diagnosis was pneumonia. Most cases were less than 2 years old, making up 81.69% (58/71), amony them, 13 months-18 months age andor = 24 months groups have the highest incidence rates, the incidence rate is 33.33%.Human rhinovirus happened in spring and winter seasons, especially the infants who were under 2 years are the main infection groups, the important symptoms are lower respiratory infections such as pneumonia, bronchitis and bronchiolitis et al. Human rhinovirus is seasonal and contagious, spreads fast, so protective measures in hospitals should be prepared to avoid cross-infection.

To analyze the genetic characterizations of wild type measles viruses caused the measles outbreak in Xinjiang.Vero/Slam cell were used for measles viruses isolation from the specimens collected from measles outbreaks patients. Fragment of 676 nucleotide acids of the carboxylend of nucleoprotein gene were amplified using reverse transcription-polymerase chain reaction (RT-PCR) method and then the PCR products were directly sequenced and analyzed. Phylogenetic tree was constructed based on 450 nucleotide acids of the N-terminus of nucleoprotein gene, and homological analysis was performed at nucleotide acid level.11 measles viruses were sequenced and all belonged to H1a subgenotype. The nucleotide difference was 0-0.2% between 11 Xinjiang isolates. And the nucleotide difference was 2.2%-2.4% between Xinjiang isolates and H1 genotype reference strain.The Measles viruses causing the measles outbreak in Xinjiang were H1a subgenotype.

To study genetic characteristic of fusion gene (F gene) of wild type measles virus circulated in 2006, analysis the variation regular of F gene through comparing with measles virus of 1999-2003.9 representative strains, which isolated from 8 provinces in 2006, were selected. The whole F gene were amplified by reverse transcript-polymerase chain reaction and were sequenced, then they were compared with the nucleotide acid sequences of Chinese measles vaccine strains and the representative strains of 1999-2003, finally the phylogenetic analysis were conducted.Among 9 representative strains circulated in China in 2006, the homology of nucleotide acid and amino acid were 98.5%-99.8% and 99%-100% respectively, and they were 95.4%-96.2% and 96.7%-97.2% respectively compared with Chinese Vaccine strains, the homology of nucleotide acid and amino acid were 97.9%-99.7% and 98.1%-100% respectively compared with strains of 1999-2003, In F gene, the 3 glycosylation site (aa32, aa64 and aa70), aa112 and aa195, which played the important role to viral fusion, that have no changes.The F genes of measles virus of china in 2006 had no significant variation, the genetic diversity was not predominance comparing with strains of 1999-2003, and important functional sites in Fusion protein didn't change, the measles epidemic was not correlate with the F gene variation. Because F protein play an important role in Measles viruses, it needs to perform the routine monitoring of F gene, which benefit for understanding of epidemic features and genetic variation pattern of wild type Measles viruses.

To evaluate the running status of Measles laboratory network of China (Hong Kong, Macao and Taiwan were excluded) in 2008.To analyze the database of Measles laboratory network surveillance of the year 2008, and the database of serologic and virologic surveillance of National laboratory for Measles in Chinese Centers for Disease Control and Prevention(CCDC), then the indicators of the running of Measles laboratory network of China were analyzed.1, serologic surveillance: 107,160 Measles sera samples were collected between Feburary and September of 2008, and the collection rate was 77.93%; 53 778 samples were qualified and positive for IgM, the positive percentage was 50.2%. 2, Virologic surveillance: 287 Measles viral isolates were isolated by 18 provincial Measles laboratories in 2008, all were certified as H1a genotype, H1a genotype was still the predominant genotype circulating in China; 29 Rubella viral isolates were isolated by 4 provincial Measles laboratories in 2008, all belonged to 1E genotype. 3, Laboratory quality control: National laboratory for Measles passed the proficiency test and on-site review in 2008; all provincial Measles laboratories passed the sera samples recheck and proficiency test hold by National laboratory for Measles in 2008; Tianjin, Shanxi, Shandong, Zhejiang, Jilin, Hubei, provincial Measles laboratory passed the on-site review by WHO.The running status of Chinese Measles laboratory network was good in 2008, and good laboratory quality control system was also set up, methods such as specimens collection, serologic detection, cell culture and viral isolation, etc, were standardized, and applied to Chinese Measles laboratory network, and it provided important scientific basis for eradication Measles in the year of 2012.

Varicella is an acute, highly contagious respiratory and caused by Varicella-zoster virus (VZV). After the primary infection, VZV induce Varicella, then the virus remains dormant in sensory nerve roots in life. Upon the reactivation, shingles can occur. Most of patients can recovery very soon, but kinds of potentially severe complication can induce death. Nowadays the VZV vaccination isn't included in the expanded immunization program in our country. Varicella outbreaks take place at times in the middle of children of our country. The paper offers concise description about pathogen characteristics, diagnosis methods, clinical character, epidemiology knowledge and strategy of prevention.

To study the genotype and gene characterization of measles wild viruses circulated in Jilin provinces, and to provide scientific evidences for setting down controlling and preventing strategy and measures.38 strains of measles virus isolated in 2001-2006 were genotyped by RT-PCR-RFLP, some strains of measles virus in Jilin province were chosen for the phylogenetic analysis and for the homology analysis of nucleotide and amino acid sequences.All the 38 strains of measles virus were identified as H1 genotype by RT-PCR-RFLP, and 29 strains of them were identified further as H1 a by sequence analysis. The homology of nucleotide was 88.0%-89.4% and the homology of amino acid was 91.8%-92.7% .The average diversity was less than 1.4%.The measles virus of H1a genotype was the circulating virus within recent years in Jilin province. There were the same measles virus strains circulating and transmitting at different years and also the different H1a measles virus strains co-circulating at the same year. There were the same transmission chain caused by the same measles virus with other provinces.

A new simple RT-LAMP method was applied to detect measles virus nucleic acid and compared with nest-RT-PCR.Compare the detection rate of the RT-LAMP method with that of nest-RT-PCR by detecting measles virus nucleic acid from measles virus and clinical samples.The nucleic acid positive rates of all 23 strains of measles virus are all 100% by the two methods. But to the detection of 18 clinical samples which are negative in measles isolation, the nest-RT-LAMP showed 56.52% positive rate of nucleic acid of measles virus and nest-RT-PCR showed 47.83%.RT-LAMP is more sensitive than nest-RT-PCR.

Human bocavirus (HBoV) is a parvovirus recently found to possibly cause respiratory tract disease in children and adults. This study investigated HBoV infection and its clinical characteristics in children younger than five years of age suffering from acute lower respiratory tract infection in Beijing Children's Hospital.Nasopharyngeal aspirates were collected from children suffering from acute lower respiratory tract infection during the winters of 2004 to 2006 (from November through the following February). HBoV was detected by polymerase chain reaction amplification and virus isolation and the amplification products were sequenced for identification.HBoV infection was detected in 16 of 333 study subjects. Coinfections with respiratory syncytial virus were detected in 3 of 16 HBoV positive patients with acute lower respiratory tract infection. The median age for HBoV positive children was 8 months (mean age, 17 months; range, 3 to 57 months). Among the HBoV positive children, 14 were younger than 3 years old, 9 were younger than 1 year old and 7 were younger than 6 months. These 16 positive HBoV children exhibited coughing and abnormal chest radiography findings and more than 60% of these children had wheezing and fever. Ten children were clinically diagnosed with pneumonia, 2 bronchiolitis, 2 acute bronchitis and 2 asthma. One child died.HBoV was detected in about 5% of children with acute lower respiratory infection seen in Beijing Children's Hospital. Further investigations regarding clinical and epidemiologic characteristics of HBoV infection are needed.

57 rubella virus strains were isolated using Vero cell line or Vero/SLAM cell line from patients' throat swabs during rubella outbreaks and sporadics in 10 provinces of China from 2003 to 2007. Fragments of 1107 nucleotides of E1 genes of the isolates were amplified by RT-PCR, the PCR products were directly sequenced and analyzed. The phylogenetic analysis based on 739 nucleotides showed that out of 57 Chinese rubella virus strains, 55 belong to a distinguish branch of 1E genotype when comparing with 1E genotype rubella strains from other countries, and the other 2 Chinese rubella virus strains belong to 2B genotype. Most of the nucleotide mutations of 57 rubella viruses were silent mutations, and the amino acid sequences were highly conserved. Except one amino acid change (Thr212 --Ser212) in two rubella viruses at the hemagglutination inhibition and neutralization epitopes, there had no change found at the important antigenic epitope sites of the other rubella viruses. 1E genotype rubella viruses isolated from 10 provinces of China from 2003 to 2007, and two imported 2B genotype rubella viruses from Vietnam suggested that 1E genotype was the predominant genotype in this period of time. The rubella virus genotypes circulated during 2003 to 2007 were different from that circulating during 1979 to 1984 and 1999 to 2002, the rubella prevailed in recent years was mainly caused by 1E genotype rubella viruses with multi-transmission routes.

To evaluate the efficacy and safety of video-assisted thoracoscopic (VATS) esophagectomy in the treatment of esophageal cancer.From June 2004 to October 2007, video-assisted thoracoscopic esophagectomy was performed in 36 patients, including 29 men and 7 women with median age of 58.9 years old. The cancer located at upper segment in 5 cases, middle 25 cases and lower 6 cases. VATS approach was used to mobilize the intrathoracic esophagus and stomach was mobilized by open approach. Esophagogastric anastomosis was performed in the left neck.The mean operative time was 250 minutes (190-330 min) and average time of VATS was 70 minutes. The mean hospital stay was 8.7 days. Mean lymph node harvest was 14.3 nodes. Post-operative complications occurred in 11 patients(30.6%), but no perioperative death occurred.Video-assisted thoracoscopic esophagectomy is technically feasible and safe with lower morbidity and shorter hospital stay as compared to open procedure, and may replace the open esophagectomy in selected patients.

To investigate the value of multislice spiral computed tomography (MSCT) in demonstrating the relationship between bronchus and peripheral lung cancer.We prospectively performed volumetric targeted scans of 0.5 mm collimation with MSCT and reconstructed images of multiplanar reconstruction (MPR), curved multiplanar reformations (CMPR) and surface shaded display (SSD) in 53 peripheral lung cancers. The results were compared with macroscopic and microscopic specimens.(1) The third- to seventh-order branches of the bronchi were clearly shown in all patients by the designed protocol. CT demonstrated the tumor-bronchus relationship in 29 (96.7%) adenocarcinomas and 13 (76.5%) squamous-cell carcinomas. Statistic analysis showed that there was no significant difference between the two groups (chi(2) = 2.8, P0.05). (2) The tumor-bronchus relationship was identified as four types with MSCT. Type I: bronchus was obstructed abruptly by the tumor, type II: bronchus penetrated into the tumor with tapered narrowing and interruption, type III: bronchus lumen shown within tumor was patent and intact, type IV: bronchus ran at the periphery of the tumor with intact or narrowed lumen. (3) Type I was shown in 31 of 53 (58.5%) tumors with squamous-cell carcinoma slightly more common than adenocarcinoma. Type II and type III were seen equally in 8 of 53 (15.1%) tumors which occurred only in adenocarcinomas. Type IV was seen in 15 of 53 (28.3%) tumors with adenocarcinoma being slightly more frequent than squamous cell carcinoma. (4) The tumor at the fourth-order bronchus was more common in squamous cell carcinoma, whereas that at the fixth-order bronchus was more likely in adenocarcinoma.Volumetric targeted scan of ultra-thin section with MSCT and followed by MPR, CMPR and SSD reconstruction can greatly improve the manifestation of the bronchioles and accurately demonstrate the patterns of tumor-bronchus relationship, thereby reflecting pathologic changes to some extent.

Background Lung ischemia–reperfusion injury remains a significant problem after lung transplantation. Caspase-mediated apoptotic pathways play an important role in lung ischemia–reperfusion injury, and caspase-3 is presumed to be the "effector" protease in the apoptotic cascade. Silencing gene expression of caspase-3 by short hairpin RNA (shRNA) can downregulate the caspase cascade. Therefore, we evaluated the therapeutic efficacy of caspase-3 shRNA in a rat model of lung ischemia–reperfusion injury. Methods Lung ischemia–reperfusion injury was induced in rats by clamping the hilum of the left lung for 1 hour. In vivo delivery of caspase-3 shRNA was performed by intratracheal administration 48 hours before ischemia. As controls, animals received either scrambled shRNA or RNase-free 5% dextrose in water solution. Real-time polymerase chain reaction, Western blotting, and immunohistochemistry were used to assess the gene silencing efficacy. The therapeutic effects of shRNA were evaluated by lung function analysis and the ratio of wet/dry weight. Results In this study, we have shown that ischemia–reperfusion injury is associated with an increased level of lung caspase-3 messenger RNA. Animals treated with caspase-3 shRNA showed a significant downregulation in lung expression of caspase-3 at transcripts and protein levels. Lung function was protected by caspase-3 shRNA therapy, inasmuch as levels of partial pressure of oxygen and carbon dioxide were significantly increased and reduced, respectively. Conclusions In summary, we have demonstrated the therapeutic potential of shRNA to knock down the expression of caspase-3 and prevent lung apoptotic injury. Our findings may have some potential therapeutic relevance for treating lung ischemia–reperfusion injury after transplantation.