OBJECTIVETo explore the relationship between epidermal growth factor receptor (EGFR) gene expression and radiosensitivity of non-small-cell lung cancer (NSCLC) cells.

METHODSEGFR sequence-specific double-stranded RNA (dsRNA-EGFR) was chemically synthesized. NSCLC cell line SPC-A1 was transfected with dsRNA-EGFR formulated with Lipofectamine 2000. Western blot and real-time PCR were used to determine the EGFR mRNA and protein expression, respectively. Colony inhibition test was adopted to observe the radiosensitizing effect. To establish the nude mouse tumor models, calculate the tumor growth inhibition rate and make the tumor growth curve by measuring its size and weight.

RESULTSEGFR mRNA levels were 1.51 ± 0.22, 1.38 ± 0.15 and 0.45 ± 0.11 in the control group, dsRNA-unrelated group and dsRNA-EGFR group, respectively (F = 482.7, P < 0.01). The contents of EGFR protein were 2340.87 ± 10.99, 2231.85 ± 35.66 and 832.03 ± 39.13 in the control group, dsRNA-unrelated group and dsRNA-EGFR group, respectively (F = 263.3, P < 0.05). Compared with the control group, dsRNA-EGFR sequence specifically decreased the expressions of EGFR mRNA by 70.2% and EGFR protein by 64.5%. The colony inhibition rates of the control group, dsRNA-unrelated combined with radiotherapy group and dsRNA-EGFR combined with radiotherapy group were 9.3%, 12.5% and 65.5%, and the tumor growth inhibition rates were 21.3%, 24.4% and 64.2%, respectively. The combination of dsRNA-EGFR and radiotherapy significantly inhibited the tumor growth in vitro and in vivo.

CONCLUSIONSDsRNA-EGFR shows an apparent inhibitory effect on the expression of EGFR mRNA and protein of NSCLC cells, effectively inhibit the tumor growth in vivo, and enhance the radiosensitivity of NSCLC.

Adenocarcinoma ; metabolism ; pathology ; radiotherapy ; Animals ; Cell Line, Tumor ; Cell Proliferation ; radiation effects ; Humans ; Lung Neoplasms ; metabolism ; pathology ; radiotherapy ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Double-Stranded ; genetics ; RNA, Messenger ; metabolism ; Radiation Tolerance ; Random Allocation ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Transfection ; Tumor Burden ; radiation effects

Spillage of cyst contents during surgical operation is the major cause of recurrence after hydatid cyst surgery. Instillation of a scolicidal agent into a hepatic hydatid cyst is the most commonly employed measure to prevent this complication. SB202190 is a pyridinyl imidazole derivative and is known to be a specific inhibitor of p38 MAPK. In the present study, the scolicidal effect of SB202190 was investigated. Freshly isolated Echinococcus granulosus protoscolices were subjected to SB202190 treatment (10, 20, 40, and 80 microM), and the effects on parasite viability were monitored by trypan blue staining. Corresponding effects were visualized by scanning and transmission electron microscopy. Dose-dependent protoscolex death within a few days of SB202190 treatment was observed. Although the in vitro scolicidal effect of SB202190 was satisfactory, the in vivo efficacy of this drug and also possible side effects remain to be further investigated.
Animals ; Anthelmintics/*pharmacology ; Dose-Response Relationship, Drug ; Echinococcus granulosus/*drug effects/ultrastructure ; Imidazoles/*pharmacology ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Parasitic Sensitivity Tests ; Pyridines/*pharmacology ; Survival Analysis

Objective To observe the blood biochemical and histological changes before and after pancreas freezing, to provide evidence for cryosurgery for pancreatic cancer. Methods Fifteen healthy pigs were divided into deep frozen group (n = 5), shallow frozen group (n = 5), non-frozen group (n = 3) and normal group (n = 2). After anesthesia and Iaparotomy, a probe of the Argon-Helium Surgical System was inserted into the pancreas, 100% and 10% argon output power were used in deep and shallow frozen group, respectively;and the temperature were - 130 ～ - 140℃ and - 110 ～ - 120℃, respectively;which results in an ice-ball with 15 ～ 20 mm in diameter. Then helium gas was inputted to increase the temperature to 10 ～ 20℃ for three minutes;then the whole process was repeated. A probe was inserted into the pancreas in the non-frozen group only and only laparotomy was performed in non-grozen group normal group and normal group. Serum amylase, IL-6, CRP levels before and after the experiment was determined;the pigs were sacrificed at day 7 and the pancreas was harvested for light microscope and electron microscope examination. Results The frozen pancreatic tissue became pitchy necrosis zone, and it could be distinguished from non-frozen tissue;there were obvious tissue necrosis in the center and para-center of frozen area, and the ultra-structure were destroyed and disappeared, mitochondria degranulation and rough endoplasmic reticulum degrannlation were observed. Serum amylase was elevated in 13 (86.7%) pigs and most returned to normal at 6th day. Serum IL-6 was slightly elevated in 5 (33.3%) pigs. There was no significant difference among all the groups in term of serum CRP. All the pigs were alive until the time of sacrifice. Conclusions Cryosurgery has affirmative fatal ablative effects on pancreatic tissue, and it is safe with no serious complications.

OBJECTIVE: To investigate the method and clinical effect of the expandable intramedullary nails on fractures of extremities. METHODS: Nineteen cases of extremities long tubular bone fractures were treated with Fixion expandable intramedullary nails. RESULTS: Nineteen cases were followed up for 4-18 months,all cases healed without any complications. CONCLUSION The application of expandable intramedullary nails in the treatment of extremity fractures has the advantages of little trauma, simple operation, rigid fixation and high healing rate. It is a good treatment for fractures of extremities.
Adolescent ; Adult ; Bone Nails ; Extremities ; injuries ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Intramedullary ; instrumentation ; methods ; Fractures, Bone ; surgery ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult

Objective To explore the effect of gene mutations of arsenic transport proteins-muhidrug resistance-associated proteins(MRP1 and MRP2)on phenotype of endemic arsenic poisoning.Methods Two hundreds and thirty-nine rural residents in 3 villages of Shuocheng Region,Shanxi Province were interviewed and examined by simple random sampling who had been lived there for 20 yearn at least.All the objects were divided into two groups on the basis of clinical examination with"The Standard Diagnosis of Endemic Arsenic Poisoning" (WT/S 211-2001):subjectives with skin lesion as a arsenic poisoning group and without skin lesion as a control group. One hundred and ninety-three blood samples were collected from each participanL Seventy-five arsenic poisoning cases and 118 controls were detected the gene mutations in the 2,17,23 exons of M RPI and the 10,18,31 exons of MRP2 by PCR-single strand conformation polymorphism (PCR-SSCP) and compared by multivariate Logistic regression model. Results Seventy-five cases and 164 controls underwent questionnaires. Age[ (58.85±11.26) vs (45.73±11.92),OR = 3.378,P ＜ 0.05],gender[male,57.3%(43/75)vs 27.4%(45/164),OR = 3.553,P＜ 0.01 ],smoking[46.7%(35/75) vs 21.3%(35/164),OR = 3.225,P ＜ 0.01 ],drinking[ 17.3%(13/75) vs 8.5% (14/164),OR = 1.836,P ＞ 0.05],vegetable and fruit intake[5.3%(4/75) vs 9.1%(15/164),OR = 0.560,P ＞ 0.05],egg and meat intake[34.7%(26/75) vs 30.5%(50/164),OR = 1.210,P ＞ 0.05],exposure of pesticide [41.3%(31/75) vs 29.3%(48/164),OR = 1.864,P ＜ 0.05] were tested by Logistic regression model. There was no gene mutation detected in the 23 exon of MRP1 and the 18 exon of MRP2. The gene mutations frequencies of the 2 exons of MRP1 in arsenic poisoning and control groups were 8.00% (6/75) and 5.93% (7/118),respectively;they were 13.33%(10/75) and 8.47%(10/118) of the 17 exons of MRP1,respectively;they were 22.67%(17/75) and 18.64%(22/118) of the 10 exons of MRP2,respectively;they were 5.33%(4/75) and 2.54%(3/118) of the 31 exons of MRP2,respectively. There was no significant difference between two groups(x2 = 0.312,1.165,0.460, 2.794,respectively,all P ＞ 0.05). After age,gender,smoking,drinking,nutritional level and exposure of pesticide being adjusted by multivariate Logistic regression model,there was no significant difference between two groups (OR = 0.803,1.892,2.388,1.098,respectively,all P ＞ 0.05). Conclusions The gene mutations of 2,17,23 exons of MRPI and the 10,18,31 exons of MRP2 may have no effect on the phenotype of endemic arsenic poisoning.

OBJECTIVETo investigate the effect of the protocol recommended by NCCN-2007 on the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC) in China.

METHODSNCCN protocol consists of identifying HNPCC characteristics according to the revised Bethesda Guidelines,genetic counseling with immunohistochemistry and finally genetic testing. Four hundred and nineteen patients diagnosed as colorectal cancer from January 2002 to February 2006 were selected. The hMLH1 and hMSH2 immunostaining were implemented for 90 patients who fulfilled the revised Bethesda Guidelines, in whom 8 patients fulfilling the Amsterdam II (Criteria were classified as group A and the other 82 patients as group B. The frozen tissues were collected from patients who showed loss of hMLH1 or hMSH2 protein expression, then RNA was extracted, and RT-PCR and cDNA sequencing were adopted to detect the germline mutations of hMLH1 and hMSH2.

RESULTSTumor tissues from 18 patients showed loss of hMLH1 or hMSH2 protein expression (5 patients in group A and 13 in group B). Finally, 21 patients(8 in group A and 13 in group B showed loss expression of MMR protein) were diagnosed as HNPCC, including 2 cases of hMLH1 and 1 case of hMSH2 mutations. These 3 cases with cDNA mutations did not fulfill the Amsterdam II( Criteria, and were finally diagnosed as HNPCC.

CONCLUSIONThe protocol recommended by NCCN-2007 offers a useful approach to identify HNPCC patients,and reduces the possibility of missed diagnosis of HNPCC.

Adult ; Aged ; Aged, 80 and over ; Base Pair Mismatch ; China ; Colorectal Neoplasms, Hereditary Nonpolyposis ; diagnosis ; genetics ; Female ; Gene Deletion ; Genetic Testing ; methods ; Guidelines as Topic ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo compare the short-term efficacy and safety between complete mesocolic excision (CME) and traditional radical resection in colon cancer.

METHODSBetween January 2008 and August 2011, 92 patients undergoing elective open surgery for colon were included in the study. CME was performed in 54 patients in the period from November 2009 to August 2011. The other 38 patients underwent traditional radical resection from January 2008 to October 2009. Short-term outcomes were compared between the patients of two different time periods.

RESULTSLymph nodes retrieved in the CME group (22.2 ± 8.0) were significant more than that in the control group (18.6 ± 4.7)(P<0.05). In patients with stage III cancer, CME group was associated with higher lymph node counts (23.8 ± 7.6 vs. 16.7 ± 3.6, P<0.01), however, there were no significant differences for those with stage I and stage II cancer (P>0.05). The number of positive lymph nodes and metastatic lymph node ratio (LNR) for stage III patients in two groups were not significantly different (P>0.05). There were no differences in operation time, time to first bowel movement, hospital stay, and postoperative complications between the two groups (P>0.05). However, intraoperative blood loss in the CME group was significantly reduced (median, 100 vs. 115 ml, P<0.05).

CONCLUSIONSCME can achieve en-bloc resection of the tumor and mesocolon, and have optimal lymph nodes harvest. Despite wider resection extent with CME technique, the surgical risk and postoperative complications are not increased and the short-term efficacy is good.

Colectomy ; methods ; Colonic Neoplasms ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Mesocolon ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo explore the clinical and pathological features and the diagnosis of childhood Alport syndrome (AS).

METHODSA retrospective analysis was performed on clinical data of 91 children with AS.

RESULTSHematuria was observed in all 91 patients, of whom 86 were accompanied with proteinuria. Sixty-one children with X-Linked AS (XL-AS) had positive family history. Renal biopsy was performed on 82 children. Mild to moderate mesangial proliferation was observed in 74 cases. Small amounts of immune complexes deposits in the glomerular mesangial area were observed in 48 cases. Glomerular basement membrane (GBM) attenuation, thickening and layering were observed in 53 cases by electron microscopy (EM). In 63 cases receiving renal tissue type IV collagen α3 and α5 chain immunofluorescence detection, 58 were diagnosed with AS, including 53 cases of XL-AS and 5 cases of autosomal recessive AS. In 91 cases of AS, 58 were diagnosed as AS by renal tissue type IV collagen α3 and α5 chain immunofluorescence, 21 were diagnosed by EM, one was diagnosed by skin biopsy, and 12 were diagnosed by gene detection. Six novel mutations of COL4A5 gene were found. Forty-five cases were misdiagnosed before the diagnosis of AS. Forty-one of the 45 cases received steroids and/or immunosuppressant therapy.

CONCLUSIONSThe clinical manifestations and pathological changes are not specific in children with AS, resulting in a higher rate of misdiagnosis. Typical lesions of GBM under EM are only observed in a part of patients. There is a high novel mutation rate of COL4A5 in the detected AS children.

Child ; Child, Preschool ; Collagen Type IV ; genetics ; Diagnostic Errors ; Female ; Glomerular Basement Membrane ; pathology ; Humans ; Male ; Nephritis, Hereditary ; diagnosis ; genetics ; pathology ; Retrospective Studies

OBJECTIVETo investigate the pathologic bacterial distribution and their antibiotic resistance in infants aged from 1 to 3 months with lower respiratory tract infection, so as to provide instructions for clinical application of antibiotics.

METHODSInduced sputum was extracted from 622 cases of hospitalized infants aged from 1 to 3 months with lower respiratory tract infection between January 2013 and December 2013, and microbial sensitivity test was performed with agar diffusion sensitivity test.

RESULTSA total of 379 (60.9%) strains of bacteria were isolated from induced sputum in the 622 infants. The Gram-negative strains were detected in 325 strains (85.8%), and the Gram-positive strains were found in 50 strains (13.2%) in the 379 strains. The others were Fungal strains (4 strains, 1.1%). The Gram-negative bacteria included Escherichia coli (31.1%) and Klebsiella pneumoniae (18.2%), with extended-spectrum β-lactamases (ESBLs) production of 48.3% and 52.2% respectively. The average rate of antibiotic resistance for ESBLs-producing bacteria was 53%. ESBLs-producing bacteria were highly resistant (100%) to ampicillin and cefotaxime, but sensitive to carbapenems. Staphylococcus aureus (10.0%) was the dominant bacteria in Gram-positive bacteria. A lower proportion of methicillin-resistant Staphylococcus aureus (1.8%) was observed, however the resistance rate of methicillin-resistant Staphylococcus aureus to β-lactam antibiotics were 100%.

CONCLUSIONSEscherichia coli and Klebsiella pneumoniae are the main pathogenic bacteria causing lower respiratory tract infection in infants aged from 1 to 3 months. ESBLs-producing bacteria accounted for over 48%, and the antibiotic resistance rate were more than 53% in these infants. These results provide a basis for the first empirical clinical use of antimicrobial in infants with lower respiratory tract infection.

Drug Resistance, Bacterial ; Escherichia coli ; isolation & purification ; Female ; Humans ; Infant ; Klebsiella pneumoniae ; isolation & purification ; Male ; Respiratory Tract Infections ; drug therapy ; microbiology ; Sputum ; microbiology

OBJECTIVETo develop an alternative method for assessment of gene delivery systems in vivo.

METHODSMouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase (Gluc) expression cassette. After implantation of these cells into recipient mice, the expression of Gluc was detected in whole blood or plasma collected.

RESULTSAs little as 10 muL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer. And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice.

CONCLUSIONSGluc may be useful as an in vivo reporter for gene therapy researches, and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.

Animals ; Arecaceae ; enzymology ; Cell Line ; Gene Transfer Techniques ; Genes, Reporter ; Humans ; Luciferases ; genetics ; Mice