Objective: To develop a PCR method for detecting S treptococcus sobrinus (S.s) in conventional culture. Method: A pair of specific primers were designed from the dexA gene of S.s , the genome DNA of 12 strains of Mutans Streptococci (8 serotypes from a ～h) and 23 species of the bacteria which were commonly found in oral cavity wer e tested by PCR amplification, the PCR products were identified by electrophore sis. Result: Only S.s of serotype d and g could produ ce 277 bp DNA fragments, and the PCR method could detect less than 1 000 copies of S.s. Conclusion: PCR method is specific and sen sitive in the detection of S.s.

Objective To explore the fluctuation of cytokine mRNA expression level in a novel T-cell-mediated immune hepatic fibrosis model induced by repeatedly injections of Concanavalin A in BALB/c mice. Methods BALB/c mice were divided into different groups. Model group mice were injected weekly up to 20 weeks with Concanavalin A (15mg/kg), via retro-orbital venous plexus under ether anesthesia. Normal control group mice were treated in the same manner weekly with normal saline. Twenty-four hours after Concanavalin A challenge at 1, 5, 12 and 20 week, 8 mice from each time were killed by cervical dislocation, repectively. The livers of different group were excised and fixed in 10% formalin for HE staining and Gomori Ag staining or frozen in optimal cutting temperature (O.C.T.) media in liquid nitrogen for immunohistochemical staining for CD4 +T or CD8 +T cell. After extracting total RNA from liver tissues, IL-2, IL-4, IL-10 and transforming factor ?1 messenger RNA were amplified by reverse transcription polymerase chain reaction. PCR products were electrophoresed on agrose containing ethidium bromide and visualized under ultraviolet light. Densitometric RT-PCR data were standardized with ?-actin signals. Results The histological change of HE staining and Gomori Ag staining indicated the fibrogenesis in model group mice. Immunohistochemical staining for CD4 + or CD8 + T cell indicated that the infiltrating lymphocytes in liver parenchyma were mainly CD4 +T lymphocytes. IL-2 mRNA expression level only increased after the first injection of Concanavalin A. The expression levels of IL-4, IL-10 and transforming growth factor ?1 mRNA significantly increased over the whole experiment period as compared with control group. Conclusions Repeated administration of Concanavalin A can induce T-cell-mediated immune hepatic fibrosis model in BALB/c mice. The expression levels of IL-4, 10 and TGF-?1 increase over the whole experiment period and may play an important role in creating mouse fibrotic model.

Objective To summarize the therapeutic experience on malignancy patients complicated with hyperthyroidism.Methods Clinical date of 10 cases of surgical malignancy complicated with hyperthyroidism admitted from May 2004 to May 2010 in Tianjin Medical University General Hospital were analyzed retrospectively.Results Four cases were treated by subtotal thyroidectomy before radical operation for cancer.Radical operation for cancer was performed on 6 patients after clinical symptoms of hyperthyroidism were controlled by perioperative antithyroid agents.Postoperatively 3 patients complicated with clinical manifestations similar to thyroid crisis.There was no postoperative mortality.Chemotherapy was given to 10 patients,and 1 patient was discontinued for chemotherapy caused leucopenia.Conclusions Hyperthyroidism should be controlled by surgery or antithyroid agents before patients of malignant diseases could proceed with radical surgery.Proper preoperative medication and effective postoperative management can reduce operation risk and help the patients get through the perioperative period safely.

At present, there are many methods for genotype of hepatitis C virus , but not a gold standard. In order to establish the rationale for genotypic determination of optimal region sequence, fifteen complete genome sequences of hepatitis C which had been given the annotation about every region and derived from different country were downloaded from GenBank. Phylogenetic trees on 5' UTR, core, E1, E2 and NS5B region were established. The results demonstrated that genotyping group was not all correct on 5' UTR region while genotyping groups were wholly correct on core, E1, E2 and NS5B region. Comparing phylogenetic distances on core, E1, E2 and NS5B region with that on complete genome sequence demonstrated that the NS5B area was the best genotyping region instead of the complete genome sequence. In addition, analysis of the molecular evolution on each core region could supply some clues for creating novel genotyping method based on PCR-RFLP.

Objective To investigate the adjunctive therapeutic effects and safety of intravenous immunoglobin (IVIG) for treating pneumonia following kidney transplantation.Methods Sixteen cases of pulmonary infection after kidney transplantation were divided into two groups.Twenty-eight cases were subjected to IVIG therapy (0.2 g·kg-1 ·day-1) for 7-10 days besides the standard specific anti-bacterial,anti-fungal,and anti-virus treatment and regular immunosuppressive regimen with dose adjustment (IVIG group),and the control group was only treated with standard specific anti-pathogen therapy.The incidence and mortality ofsevere pulmonary infection,levels of serum IgG,T lymphocyte subsets,and creatinine in the two groups were observed.Results The effective power of IVIG group and control group was 100 ％ and 93.75 ％ (P＜0.05).The incidence of severe pneumonia in IVIG and control groups was 0 and 12.5％,respectively (P＜0.05),with the mortality being 0 and 6.25％,respectively (P＜ 0.05).The levels of serum IgG were significantly increased in IVIG group as compared with that before treatment and in control group.There were no significant adverse reactions associated with IVIG infusion.Conclusion As an adjunctive therapy,IVIG treatment for pulmonary infection can reduce the incidence of severe pulmonary infection and mortality after kidney transplantation,further increase the survival rate of patients after kidney transplantation.

Objective To investigate the effects of CNTN-1 on the invasion and migration of human esophageal cancer EC9706 cells and the possible mechanism.Methods The expression of CNTN-1 in human esophageal cancer EC9706 cells was measured by qPCR and Western blot.After transfection with CNTN-1 siRNA or CNTN-1, the cells were divided into control group, scrambled siRNA group, CNTN-1 siRNA group, pcDNA3.1-vector group and pcDNA3.1-CNTN-1 group.Cell proliferation, invasion and migration were respectively analyzed by BrdU assay and Transwell test.The expression of MMP-2 and MMP-9 were detected by qPCR and Western blot.Results The mRNA and protein expression of CNTN-1 were significantly upregulated in EC9706 cells.Compared with control, cell proliferation, invasion and migration, as well as the expression of MMP-2 and MMP-9 were significantly decreased by CNTN-1 siRNA, while they were increased by CNTN-1 overexpression (P<0.05).ConclusionsCNTN-1 can influence the invasion and metastasis of esophageal cancer cells through the regulation of the expression of MMP-2 and MMP-9.

Background and purpose: Previous studies have confirmed that the expression of leucine-rich repeat-containing 3B (CLRRC3B) was significantly decreased in different human cancers, which was also associated with the migration and invasion of cancer cells. The aim of this study was to explore the potential mechanism of LRRC3B in the development of esophageal cancer. Methods: The LRRC3B expression was detected in 60 cancer tissues and 60 adjacent non-neoplastic tissues by immunohistochemistry. The mRNA and protein expression of LRRC3B in Eca109 and HEECs were detected using real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot, respectively. Eca109 cells with different treatments were divided into three groups:normal group, negative control group (transfected with pCMV6 plasmid), overexpression LRRC3B group (transfected with pCMV6-LRRC3B plasmid). Transwell assay was used to measure the migration and invasion of Eca109 cells in different groups. The protein levels of E-cadherin, N-cadherin, Vimentin and p-Akt were determined by Western blot. Results: The expression of LRRC3B in esophageal cancer tissues was lower than that of non-cancerous tissues, as well as the expression of LRRC3B in Eca109 was decreased compared with that of normal esophageal epithelial cell line HEEC. Overexpression of LRRC3B significantly inhibited Eca109 cells migration and invasion, upregulated the expression of E-cadherin and decreased the expression of N-cadherin and Vimentin. Moreover, overexpression of LRRC3B significantly inhibited the phosphorylation of Akt in Eca109 cells. Conclusion: The expression of LRRC3B was decreased in esophageal cancer. Overexpression of LRRC3B can efficiently inhibit the EMT progression in esophageal cancer cells by suppressing PI3K/Akt signaling pathway.

Objective To observe the therapeutic efficacy and safety of recombinant tissue-type plasminogen activator (rt-PA) combined with edaravone on acute cerebral infarction (ACI).Methods 60 cases of ACI were divided into rt-PA group (30 cases, treated by rt-PA only) and trial group (30 cases, treated with rt-PA combined with edaravone). The nerve function deficits of patients in both groups were evaluated by European Stroke Scale (ESS) and Barthel Index (BI) before and after treatment respectively.Results The ESS scores at 21st day and 90th day after treatment in the trial group were significantly higher than those of the patients in the rt-PA group. The effective rate of patients in the trial group was significantly higher than those in the rt-PA group (P＜0.05).Conclusion The treatment of rt-PA combined with edaravone is effective and safe in patients with ACI.

In April 10, 2010, an earthquake measuring magnitude 7.1 shocked Yushu County,Qinghai province. For medical rescue, the National Earthquake Disaster Emergency Rescue Team was sent to Yushu right away. Rescue work in Yushu was faced with such difficulties as short preparative time, heavy workload, high exposure to various acute high altitude diseases (AHAD), and a number of other diseases frequently found on the cold plateau. To ensure the rescue work a success, the team took a series of measures including efficient preparative procedure, scientific and logical procedure in the emergency medical aid operations, reliable and effective handling of AHADs, along with sufficient self protection for team members.

Honeybee pupae were collected from Jilin apiaries and RNA was extracted for use as a tefnplate for amplification. Based on the complete genome sequences of black queen cell virus (BQCV) published on GenBank, we designed 10 pairs of primers to amplify genes by reverse transcription-polymerase chain reaction (RT-PCR). Using this approach, we have obtained the first complete genome sequence of a BQCV isolate in China. The genome of the isolated strain, named BQCV-JL1, is composed of 8358 nucleotides and shares between 86% and 93% homology with the complete genome sequences of the other six BQCV strains published on GenBank. ORF 1 of BQCV-JL1 is positioned between nucleot ides (nt) 546 and 4676 (4131 nt), while ORF 2 is located between nt 5750 and 8203. Between the two ORFs of BQCV-JL1 there is a short ORF, called ORF 3, between nt 4891 and 5433 (543 nt). The first functional gene ex- pression domain of the BQCV-JL1 strain is positioned between nt 546 and 5 429, encompassing both ORF 1 and ORF 3. There is an internal ribosome entry site (IRES) located before ORF 2, the last three bases of which are CCU (nt 5642-5644). These bases act as an initiation.codon facilitating the translation of ORF 2. The second functional gene expression domain of the BQCV-JL1 strain is located between nt positions 5642 and 8203. The BQCV-JL1 strain was found to share high sequence identity (93%) with the Hungary 10 genotype at the whole-genome level and analysis of the nucleotide and amino acid sequences revealed that the BQCV-JL1 strain also shows close genetic relationships with the South Korea strain, suggesting that both the BQCV-JL1 and South Korea strains may have migrated from European countries. BQCV-JL1 strain was different from the other 6 strains in dividing the nucleotides positions of QRF, which vqs because of the gene mutation.
Amino Acid Sequence ; Animals ; Bees ; China ; Genome, Viral ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; RNA Viruses ; classification ; genetics ; isolation & purification ; Viral Proteins ; genetics