OBJECTIVEThe purpose of this study was to develop RT- PCR assay for Rapidly detect and distinguish between Norovirus genogroup I and genogroup II with a pair of primers.

METHODSA pairs of primers specific to capsid prote in ORF2 gene of G I and G II Norovirus were dsigned according to the published complete genome sequence, with which the RNA of Norovirus was extracted and RT-PCR amplification. The sensitivity, specificity of the RT- PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.

RESULTSThe results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses, including rotavirus, enteric adenovirus and hepatitis A virus. The detection limit of RT-PCR assay for Norovirus G I and G II were up to 100 pg/ml and 10 pg/ml respectively.

CONCLUSIONThe RT- PCR assay provide rapid and sensitive detection of Norovirus G I and G II and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.

Caliciviridae Infections ; diagnosis ; virology ; DNA Primers ; genetics ; Gastroenteritis ; diagnosis ; virology ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods

OBJECTIVEThe purpose of this study is to investigate the osteogenic potential and possibility of combination application of mimetic osteoinductive periosteum with tissue-engineered bone.

METHODSThe three-dimensional construction of tissue-engineered bone was made by implantation of adipose derived stromal cells (ADSCs) into rhBMP-2 mediated bio-derived carrier, and mimetic periosteum was constructed by loading ADSCs into Cs-Col-beta3-TCP with rhBMP-2. 10 mm defects of right radiuses were established in adult New Zealand rabbits, group A was transplanted by tissue-engineered bone with mimetic periosteum, group B was implanted by tissue-engineered bone, and group C was implanted by mimetic periosteum, group D was transplanted by bio-derived compound bone as blank scaffold. X-ray, histology, immunohistochemistry stain, dural energy X-ray absorptiometry (DEXA) and transmission electron microscopy (TEM) examinations were performed at different periods.

RESULTSGroup A played a predominant role in process of new tissue regeneration and mature bone reconstitution, defect completely healed at 12 weeks. Group B showed primary repair, group C also existed in modeling stage. While, group D displayed retard regeneration with poor osteogenic capacity. DEXA result showed that group A had statistical significance over control group according to data of BMC and BMD ( P < 0.05).

CONCLUSIONSEnhanced osteogenic potential can be obtained by using tissue-engineered bone with mimetic osteoinductive periosteum. Defect can be healed with concord pattern of osteoinductive and osteopromotive and osteoconductive effects.

Animals ; Biocompatible Materials ; Bone Regeneration ; Bone Substitutes ; Male ; Periosteum ; Rabbits ; Radius ; pathology ; surgery ; Tissue Engineering ; methods

OBJECTIVETo analyze the feasibility of reverse island flaps pedicled with cutaneous nerve nutrient vessels in repairing the defect on distal parts of extremities.

METHODSThirty patients with tissue defect in distal extremities, complicated by exposed vessels, nerve, tendons, and/or bones ,were repaired with island flaps pedicled with neurocutaneous nutrient vessel. Among them, five cases were grafted with flaps with external forearm cutaneous nerve vessels, eleven cases were grafted with flap containing sural neural nutrient vessels in its pedicle,and 14 cases with saphenous nerve nutrient vessels in the pedicles.

RESULTSThe flaps of 28 cases survived with perfect appearance and function. In one case partial necrosis occurred because of compression on the pedicle, but the patient healed after dressing, and another case with necrosis of the edge of the flap due to infection, but also healed after skin grafting.

CONCLUSIONThe reverse island flaps pedicled with cutaneous nerve nutrient vessels which are constant in anatomy, with reliable blood supply, can be recommended because its simple operative technique, non-injurious to main blood vessels and muscles, the repair of distal defects of the extremities.

Adolescent ; Adult ; Child ; Extremities ; injuries ; Humans ; Male ; Middle Aged ; Peripheral Nervous System ; Reconstructive Surgical Procedures ; Skin ; innervation ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; innervation ; Wound Healing ; Young Adult

Postoperative radiotherapy is a major treatment for patients with maxillary sinus carcinoma. However, the irregular resection cavity poses a technical difficulty for this treatment, causing uneven dose distribution to target volumes. In this study, we evaluated the dose distribution to target volumes and normal tissues in postoperative intensity-modulated radiotherapy (IMRT) after placing a water-filled balloon into the resection cavity. Three postoperative patients with advanced maxillary sinus carcinoma were selected in this trial. Water-filled balloons and supporting dental stents were fabricated according to the size of the maxillary resection cavity. Simulation CT scans were performed with or without water-filled balloons, IMRT treatment plans were established, and dose distribution to target volumes and organs at risk were evaluated. Compared to those in the treatment plan without balloons, the dose (D98) delivered to 98% of the gross tumor volume (GTV) increased by 2.1 Gy (P = 0.009), homogeneity index (HI) improved by 2.3% (P = 0.001), and target volume conformity index (TCI) of 68 Gy increased by 18.5% (P = 0.011) in the plan with balloons. Dosimetry endpoints of normal tissues around target regions in both plans were not significantly different (P > 0.05) except for the optic chiasm. In the plan without balloons, 68 Gy high-dose regions did not entirely cover target volumes in the ethmoid sinus, posteromedial wall of the maxillary sinus, or surgical margin of the hard palate. In contrast, 68 Gy high-dose regions entirely covered the GTV in the plan with balloons. These results suggest that placing a water-filled balloon in the resection cavity for postoperative IMRT of maxillary sinus carcinoma can reduce low-dose regions and markedly and simultaneously increase dose homogeneity and conformity of target volumes.
Adult ; Aged ; Carcinoma, Adenoid Cystic ; diagnostic imaging ; radiotherapy ; surgery ; Carcinoma, Squamous Cell ; diagnostic imaging ; radiotherapy ; surgery ; Female ; Humans ; Male ; Maxillary Sinus ; surgery ; Maxillary Sinus Neoplasms ; diagnostic imaging ; radiotherapy ; surgery ; Middle Aged ; Postoperative Period ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Intensity-Modulated ; methods ; Stents ; Tomography, X-Ray Computed

OBJECTIVETo evaluate the surgical therapeutic strategy for non-small cell lung cancer (NSCLC) with (N2) mediastinal lymph node metastasis.

METHODSThe clinical data of 325 patients with N2 NSCLC treated surgically between 1961 and 1995 were analysed.

RESULTSThe over-all 5-year survival rate was 19.6%. Survival was higher in patients with radical resection than with palliative resection, with squamous-cell carcinoma than with adenocarcinoma, with sleeve lobectomy and pneumonectomy than with regular lobectomy, with 1 to 3 mediastinal metastatic lymph nodes than those over 4, and with adjuvant therapy (chiefly postoperative radiotherapy) than without. All these differences were statistically significant (P < 0.05). There was no 5-year survivor in patients with T3 or T4 tumor, nor in those with distant metastasis.

CONCLUSIONIt is suggested that surgery is the best choice for N2 NSCLC patients with T1 or T2 tumor, with non-adenocarcinoma, and with metastatic mediastinal lymph nodes less than 4 in number. Surgery is probably not a good choice in those with T3 tumor varieties. At operation, radical resection of the tumor and systematic removal of all hilar and mediastinal lymph nodes are essential for disease staging and survival improvement. Adjuvant therapy may improve long-term survival and is especially indicated in patients with residual tumor and/or metastatic mediastinal lymph nodes over 3 in number.

Adenocarcinoma ; drug therapy ; radiotherapy ; secondary ; surgery ; Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; radiotherapy ; secondary ; surgery ; Carcinoma, Squamous Cell ; drug therapy ; radiotherapy ; secondary ; surgery ; Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; radiotherapy ; surgery ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Mediastinum ; Middle Aged ; Neoplasm Staging ; Pneumonectomy ; methods ; Radiotherapy, Adjuvant ; Survival Rate

OBJECTIVETo explore the methods of the treatment and the principles of the prevention of bronchus-pleural fistula (BPF) after pneumonectomy.

METHODSThe clinical data of 15 cases of BPF after pneumonectomy in 815 lung cancer cases treated from July 1999 to June 2006 were analyzed retrospectively.

RESULTSThe occurrence rate of BPF after right pneumonectomy was 3.9% (12/310), higher than 0.6% (3/505) of left pneumonectomy (P < 0.01). The occurrence rate of BPF in cases with positive cancer residues in stump of bronchus was 22.7% (5/22), higher than 1.3% (10/793) of the cases with negative stump of bronchus (P < 0.01). The occurrence rate of BPF in the cases received preoperative radio- or chemotherapy was 5.0% (6/119), higher than 1.3% (9/696) of the cases received operation only (P < 0.05). There were no BPF occurred in the 76 cases whose bronchial stump were covered with autogenous tissues. All of the cases diagnosed as BPF were undertaken either closed or open chest drainage. Two cases were cured by thoracentesis aspiration and infusion antibiotics repeatedly. Two cases were cured by blocking the fistula with fibrin glue after sufficient anti-inflammatory treatment and hypertonic saline flushing. Six cases were discharged with a stable condition after closed drainage only. One case was discharged with open drainage for long time and 1 case was cured by hypertonic saline flushing after failure to cover the BPF using muscle flaps. Three cases died of multi-organs functional failure.

CONCLUSIONSBPF are related to the bronchial stump management and positive or negative residue of tumor at the bronchial stump. Autogenous tissues covering of the bronchial stump is a effective method for decrease the rate of BPF and especially for those patients received preoperative radio- or chemotherapy and right pneumonectomy. It should be performed for early mild cases with repeated thoracentesis aspirations or blocking the fistula with fibrin glue together with antibiotics. Chest closed drainage immediately and flushing with hypertonic saline repeatedly are effective methods for BPF.

Adult ; Aged ; Aged, 80 and over ; Bronchial Fistula ; epidemiology ; prevention & control ; therapy ; Female ; Humans ; Lung Neoplasms ; surgery ; Male ; Middle Aged ; Pleural Diseases ; epidemiology ; prevention & control ; therapy ; Pneumonectomy ; adverse effects ; methods ; Postoperative Complications ; etiology ; prevention & control ; therapy ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo assess the association between polymorphisms of protein-activated inwardly rectifying K+ channel (GIRK4) gene and insulin resistance (IR) in Xinjiang Uygur population.

METHODSA cross-sectional epidemiological survey-based case-control study was carried out, for which 1295 subjects (including 324 IR patients and 971 non-IR controls) were randomly selected. Functional region of the GIRK4 gene was sequenced for 48 randomly selected IR patients. Representative variable sites were chosen, with its association with IR assessed in 1295 Uygur subjects.

RESULTSrs11221497 variant was associated with IR in Uygur subjects under 50 years old (P=0.017 in genotype model, P=0.009 in dominant model). Subjects with dominant model of CC genotype have an OR of 1.833 (95%CI: 1.157-2.905) for IR.

CONCLUSIONGIRK4 gene polymorphisms may be associated with IR in Uygur ethnics from Xinjiang. The CC genotype of rs11221497 variant is a risk factor for IR.

Adult ; Alleles ; Case-Control Studies ; Female ; G Protein-Coupled Inwardly-Rectifying Potassium Channels ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Insulin Resistance ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Risk Factors

OBJECTIVETo study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou.

METHODSFrom 2008 to 2010, total 119 fecal specimens collected from outbreaks of acute gastroenteritis were tested for Norovirus. Partial sequence of RNA dependent RNA polymerase (RdRp) of the positive samples were amplified by RT-PCR, then the PCR production were purified, sequenced and put into phylogenetic analysis.

RESULTS50 of 119 specimens were positive for Norovirus by real-time RT-PCR. Out of those 50 Norovirus positive specimens, 9 were Norovirus Genogroup I (GI) positive, 35 were Norovirus Genogroup II (GII) positive, 6 was both Norovirus GI and GII positive. 12 PCR products for RdRp were selected for further studies on sequencing. Phylogenetic analysis revealed that the 5 GI norovirus isolates were belonged to genotype GI/2 and GI/3. Of the 7 GII norovirus isolates, 6 were belonged to genotype GII/4, 1 was belonged to genotype Glib.

CONCLUSIONNorovirus is a major cause of outbreaks of acute gastroenteritis in Huzhou and the epidemic strains of norovirus isolated from Huzhou had a high degree of genetic diversity.

Acute Disease ; China ; epidemiology ; Disease Outbreaks ; Female ; Gastroenteritis ; epidemiology ; Genetic Variation ; Humans ; Male ; Norovirus ; classification ; genetics ; Phylogeny ; RNA Replicase ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

The tumor suppressor gene p53 and p16, both of which play an important role in inhibition of tumorigenesis, are homozygously deleted in human myeloid leukemia cell line K562. To explore the inhibition of K562 cell proliferation by wild type p16 and p53 genes, both p16 and p53 genes were co-transfected into K562 cells mediated by liposome. The expression of the two genes was measured by immunocytochemical method, the cell cycle was analysed by flow cytometry, and the number of recovered viable cells was assessed after transfection. After co-transfection, the p53 and p16 positive cells were 23% and 28%, respectively. The results showed that co-transfection of p16 and p53 genes significantly inhibits cell proliferation comparing with transfection either by p16 gene or by p53 gene (P < 0.05). Expression of p16 and p53 proteins increased the cell number in G(1) phase but decreased the cell number in S phase. It is concluded that co-transfection of p16 and p53 genes has a stronger growth-inhibitory effect on K562 cell growth than that of transfection only by p16 gene or by p53 gene, may be a pathway for gene therapy in leukemia.
Cell Division ; Genes, p16 ; physiology ; Genes, p53 ; physiology ; Humans ; K562 Cells ; Plasmids ; Transfection

OBJECTIVETo investigate the effects of clinical and pathomorphological parameters on the prognosis of colon carcinoma and rectal carcinoma.

METHODSUnivariate and multivariate COX proportional hazard models were used to study the effects of the clinical and pathomorphological factors on the prognosis in 101 cases of colon carcinoma, 219 of rectal carcinoma and 137 of rectal carcinoma under curative resections.

RESULTBy using univariate analysis, we identified that lymph node metastasis and distant metastasis were the common prognostic factors for both colon carcinoma and rectal carcinoma. Smoking, deep infiltration, chemotherapy and serum albumin concentration were the uncertain prognostic factors for colon carcinoma. Signet-ring cell carcinoma, larger tumor size (>6 cm), deep infiltration, lack of radical surgery, and advanced TNM stage were the exclusive adverse prognostic factors for rectal carcinoma. Further studies showed that the adverse prognostic factors for the rectal carcinoma under curative resection included deep infiltration, lymph node metastasis, vessel invasion, less of peritumoral lymphocyte infiltration, lack of Crohn's like reactivity, high level of tumor budding, advanced TNM stage and positive urine glucose. By using multivariate analysis based on a COX proportional hazard model, it was identified that smoking, lymph node metastasis and serum albumin concentration were independent prognostic factors for colon carcinoma; advanced TNM stage, distant metastasis and palliative surgery for rectal carcinoma; and vessel invasion, lymph node metastasis and urine glucose for rectal carcinoma under curative resections.

CONCLUSIONThe various clinical and pathomorphological parameters show different prognostic value for colon carcinoma, rectal carcinoma and rectal carcinoma under curative resections.

Adult ; Aged ; Carcinoma, Signet Ring Cell ; pathology ; surgery ; Colonic Neoplasms ; pathology ; surgery ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Rectal Neoplasms ; pathology ; surgery