OBJECTIVETo explore the effect of MSH3 knock-down on sensitivity of tongue cancer cells to cisplatin.

METHODSThree small interfering RNA (siRNA) fragments targeting MSH3 CDS region were synthesized and transfected into CAL27 cells via Lipofectamine. Real-time PCR and Western blotting were used to assess the efficiency of MSH3 silencing. MTS, apoptosis staining and cell immunofluorescence assay were used to examine the cisplatin sensitivity, apoptosis and DNA repair of transfected CAL27 cells.

RESULTSs One of the 3 siRNAs was found to significantly reduce the expression of MSH3 protein in CAL27 cells (P<0.05). MTS assay showed that MSH3 silencing resulted in an significant reduction of IC50 of cisplatin from 21.32 to 13.95 µmol/L (P<0.05) and increased the apoptotic index of the exposed cells from 4.23∓1.27 to 11.32∓1.82 (P<0.05). Immunofluorescence assay demonstrated that silencing MSH3 markedly reduced the number of γ-H2AX foci.

CONCLUSIONSilencing MSH3 can significantly increase cisplatin sensitivity of tongue cancer cells, the mechanism of which involves mainly attenuation of repair of DNA double-strand damage in the cells.

Apoptosis ; Cell Line, Tumor ; Cisplatin ; pharmacology ; DNA-Binding Proteins ; genetics ; metabolism ; Gene Silencing ; Humans ; MutS Homolog 3 Protein ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Tongue Neoplasms ; drug therapy ; genetics ; Transfection

OBJECTIVEElectron microscopical study of infected cells to identify the pathogenic agent of SARS.

METHODSVero E6 cells infected with lung autopsy samples or nasopharyngeal swabs from SARS patients of Beijing and Guangzhou were inoculated. The supernatant and cultured cells exhibiting identifiable cytopathic effect (CPE) were prepared for electron microscopic study.

RESULTSExamination of CPE cells on thin-section revealed characteristic coronavirus particles within the cisternae of endoplasmic reticulum, Golgi apparatus, vesicles and extracellular space. They were mainly spherical or oval in shape, annular or dense, about 80 nm in diameter. Negative-stain electron microscopy identified coronavirus particles in culture supernatant, 80 - 120 nm in diameter, with club-shaped surface projections. Elongated, rod-, kidney- or other irregular shaped virons with the size of 100 - 200 nm by 60 - 90 nm were also found in the cultured cells infected with the lung samples from the Guangdong patients. Infectious virons entered cells by endocytosis or membrane fusion and released through a budding process.

CONCLUSIONThese data indicate a novel coronavirus as the causative agent of SARS. Most viral particles showed typical characteristics of coronavirus. The potential role of special shape viruses is expected to be further investigated.

Animals ; Cercopithecus aethiops ; Humans ; Microscopy, Electron ; SARS Virus ; ultrastructure ; Severe Acute Respiratory Syndrome ; virology ; Vero Cells

This study was intended to evaluate the effects of hypoxic exposure on gene expression and coordination of cytochrome oxidase (COX) subunits I (COX I) and IV (COX IV) encoded by mtDNA and nDNA respectively in rat cerebral cortex. Male Wistar rats were exposed to hypoxia in a hypobaric chamber simulating high altitude at 5000 m for 2, 5, 15 and 30 d. Control rats were fed outside the hypobaric chamber (the height was 300 m above sea level). Rats were sacrificed and mitochondria from cerebral cortex were isolated by differential centrifugation at each time point. COX I and COX IV proteins in isolated rat cerebral cortex mitochondria were detected by Western blot analysis and mRNA in the cerebral cortex by RT-PCR. The ratios of protein and mRNA were used to estimate the coordinative expression of two subunits. The results showed that COX I mRNA increased significantly at 2 and 5 d, and decreased to the control level at 15 and 30 d; COX IV mRNA remarkably increased at 2, 5 and 15 d, and dropped below the control level at 30 d. The mRNA ratio of COX IV to COX I reached a peak at 15 d, but showed no differences between other time points. The Western blot analysis of COX I and COX IV in isolated rat cerebral cortex mitochondria showed no obvious changes during hypoxic exposure. Our findings demonstrate that hypoxia can affect mRNA expression of COX I and COX IV and their coordination, while protein expression of both subunits are stable and coordinative. This study suggests that the expression of COX I and COX IV proteins during hypoxic exposure is coordinately regulated by post-transcriptional mechanisms.
Animals ; Cerebral Cortex ; metabolism ; Electron Transport Complex IV ; metabolism ; Gene Expression Regulation, Enzymologic ; Hypoxia ; metabolism ; Male ; Mitochondria ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the expression of mir-106b in esophageal squamous cell carcinoma (ESCC) tissue and analyze its correlation with the clinicopathologic features of ESCC.

METHODSA total of 200 fresh surgical specimens of ESCC and adjacent tissues collected between 2001 and 2007 were examined for expressions of mir-106b using real-time PCR (RT-PCR). Northern blot analysis for mir-106b was performed in 4 pairs of samples to confirm the RT-PCR results. The relationship between mir-106b expression and clinicopathological features and prognosis of the patients were analyzed.

RESULTSMir-106b was expressed at significantly higher levels in ESCC tissues than in the paired adjacent tissues. Overexpression of mir-106b was associated with lymph node metastasis, stage of TNM classification and smoking (P<0.05). The survival rate of patients with low mir-106b expression was higher than that of patients with high mir-106b expression (60 vs 37 months, P=0.024). Cox regression analysis indicated that the expression of mir-106b, lymph node metastasis and smoking were independent prognostic factors for ESCC (P<0.05).

CONCLUSIONMir-106b is overexpressed in ESCC tumors, and its overexpression is strongly associated with lymph node metastasis and a poor prognosis. Mir-106b expression is an independent prognostic factor for ESCC and can serve as a biomarker for diagnosis and prognostic evaluation of ESCC.

BACKGROUNDComputed tomography (CT)-guided transthoracic lung biopsy is a well-established technique for the diagnosis of pulmonary lesions. The objective of this study was to evaluate the diagnostic efficiency and complication rate of CT-guided lung biopsy in a Chinese population.

METHODSCT-guided cutting needle lung biopsies were performed in our institution on 1014 patients between January 2000 and October 2010. A chest radiograph was taken after the biopsy. Data about basic patient information, final diagnosis, and complications secondary to biopsy procedure (pneumothorax and bleeding) were extracted.

RESULTSThe diagnostic efficiency of CT-guided lung biopsy was 94.8%; only 53 patients did not get a final diagnosis from lung biopsy. Final diagnoses found 639 malignant lesions (63.0%) and 322 benign lesions (31.8%). Pneumothorax occurred in 131 patients and 15 required insertion of an intercostal drain. Small hemoptysis occurred in 41 patients and mild parenchymal hemorrhage occurred in 16 patients. The overall complication rate was 18.5%.

CONCLUSIONSCT-guided cutting needle biopsy of pulmonary lesions is a relatively safe technique with a high diagnostic accuracy. It can be safely performed in clinical trials.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy, Needle ; methods ; Female ; Humans ; Lung ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; methods ; Young Adult

OBJECTIVETo explore whether vimentin (VIM) mediates the activation of inflammasome in mice with EV71 infection in the central nervous system.

METHODSForty VIM knockout mice (VIM, 3 to 5 days old) were randomly divided into control group and infection group. The infection group was intraperitoneally injected with EV71 (10 TCID), while the control group was injected with PBS (10 µL); another 40 wild-type mice (WT, 3 to 5 days old) were grouped in the same manner. The general conditions of mice were observed each day. Western blotting, ELISA, and RT-PCR were used to measure the levels of IL-1β and casepase-1 in the brain or cerebrospinal fluid. The pathological changes in the cerebella and brain were observed using immunohistochemistry.

RESULTSCompared with the control group, the VIM mice infected with EV71 showed no significant changes in NLRP3, IL-1β or caspase-1 expression. The WT mice infected with EV71 showed obviously increased NLRP3, IL-1β, and caspase-1 expressions in the central nervous system. The neurons of infected VIM mice exhibited milder cell damage than the those in WT mice.

CONCLUSIONVIM mediates the activation of inflammasome and promotes brain inflammation and neuronal damage in mice with EV71 infection in the central nervous system.

OBJECTIVETo investigate the relationship between colorectal adenocarcinoma with invasive micropapillary carcinoma (IMPC) component, lymphovascular invasion, and lymph node metastasis.

METHODSOne hundred and thirty one cases of colorectal adenocarcinoma with invasive micropapillary carcinoma component were evaluated by H&E and immunohistochemical staining. The main pathological features, percentage of IMPC component, lymphovascular invasion, and lymph node metastasis were assessed and compared to 296 cases of conventional colorectal adenocarcinoma.

RESULTSThe maximum diameter of the tumors of the IMPC group was significantly lower than the conventional group. The degree of differentiation, the lymph node metastatic rate, the average number of metastatic lymph nodes, the number of cases with lymphovascular tumor emboli were significantly higher in the IMPC group (P = 0.000). The rate of distant metastasis at operation was higher in the IMPC group (14.5%) compared to the conventional group (10.8%) even though statistical significance was not achieved (P = 0.278) . In IMPC group, as the IMPC component increased, the rate of lymph node metastasis also showed an increasing trend. The rate of lymph node metastasis were 53.0% (157/296) , 67.7% (42/62) and 85.5% (59/69) " respectively" for conventional adenocarcinoma, adenocarcinoma with ≤ 10% IMPC component and adenocarcinoma with > 10% IMPC component; the difference was statistically significant (P = 0.000).

CONCLUSIONSCompared to conventional colorectal adenocarcinomas, the colorectal carcinomas with IMPC component show a significantly higher rate of lymphovascular invasion, lymph node metastasis and distant metastasis. The percentage of IMPC component in colorectal adenocarcinoma is significantly correlated to the rate of lymph node metastasis. Therefore, presence of IMPC component and degree of tumor differentiation are predicting factors of lymph node metastasis.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Cadherins ; metabolism ; Carcinoma, Papillary ; metabolism ; pathology ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Neoplasm Metastasis ; Tumor Burden ; Young Adult

OBJECTIVETo study the immunophenotype and gene rearrangement pattern of pulmonary lymphomatoid granulomatosis.

METHODSNine cases of pulmonary lymphomatoid granulomatosis, included 5 cases of open lung biopsy, 3 cases of lobectomy specimen and 1 case of autopsy, were retrospectively analyzed by immunohistochemistry, in-situ hybridization for Epstein-Barr virus-encoded RNA, immunoglobulin and T-cell receptor gene rearrangement studies.

RESULTSThe age of patients ranged from 3 to 59 years. The male-to-female ratio was 3: 6. Histologically, all cases showed lymphocytic infiltration surrounding the blood vessels and in the perivascular areas. Most of these lymphoid cells expressed T-cell marker CD3. There were also variable numbers of CD20-positive B cells. The staining for CD56 was negative. According to the WHO classification, there were 4 cases of grade I , 1 case of grade II and 4 cases of grade III lesions. Six cases had gene rearrangement studies performed and 3 of them demonstrated clonal immunoglobulin gene rearrangement (including 1 of the grade II and 2 of the grade III lesions). No T-cell receptor gene rearrangement was detected.

CONCLUSIONSPulmonary lymphomatoid granulomatosis may represent a heterogeneous group of lymphoproliferative disorders. Some of the cases show B-cell immunophenotype and clonal immunoglobulin gene rearrangement, especially the grade II and grade lesions. They are likely of lymphomatous nature.

Adult ; Antigens, CD20 ; metabolism ; CD3 Complex ; metabolism ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Humans ; Immunohistochemistry ; Lung Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Lymphomatoid Granulomatosis ; genetics ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Grading ; Pneumonectomy ; methods ; Retrospective Studies ; Young Adult

OBJECTIVETo explore the methods for constructing the digital three-dimensional model of fetal heart.

METHODSOriginal two-dimensional CT image data sets were collected from 4 abortion fetuses with fetal malformations but not heart malformation or chromosomal abnormalities. The three-dimensional fetal heart model was reconstructed using Mimics14.0 software.

RESULTSIn the reconstructed three-dimensional fetal heart, the left atrium, left ventricle, right atrium, right ventricle, the ascending aorta, the main pulmonary and their branches, the superior cava and inferior vena cava were marked with different colors, and these structures could be displayed individually or with other structures. This model also allowed three-dimensional arbitrary scaling, shifting or rotation at any angle, and the diameter of the each vessel could be measured with the software.

CONCLUSIONThe fetal heart model can be successfully reconstructed from the CT datasets using three-dimensional reconstruction software to facilitate clinical and anatomical teaching.

Female ; Fetal Heart ; anatomy & histology ; Heart Atria ; anatomy & histology ; Heart Defects, Congenital ; Heart Ventricles ; anatomy & histology ; Humans ; Imaging, Three-Dimensional ; Models, Anatomic ; Pregnancy ; Software ; Tomography, X-Ray Computed ; Vena Cava, Inferior ; anatomy & histology

OBJECTIVETo compare the medium- and long-term effect of pneumatic ballistic extracorporeal shock wave versus ultrasound-guided hormone injection in the treatment of plantar fasciitis.

METHODSThe clinical data were collected from patients with plantar fasciitis admitted to PLA General Hospital pain department from September, 2015 to February, 2017. The patients were randomly divided into ultrasound-guided drug injection group and shock wave group. The therapeutic parameters including the numerical rating scale (NRS) scores in the first step pain in the morning, American Orthopedic Foot and Ankle Society (AOFAS) Ankle Hindfoot Scale, and thickness of the plantar fascia were monitored before and at 1 week, 1 month, 3 months, and 6 months after the treatment. The recurrence rate, effectiveness, and patient satisfaction were compared between the two groups at 6 months after the treatment.

RESULTSThirty-nine patients were enrolled in shock wave group and 38 patients in ultrasound group. The NRS scores in the first step pain in the morning were lowered after treatment in both groups (P<0.05), and the scores were significantly lower in ultrasound group than in shock wave group at 1 week and 1 month (P<0.01), but significantly higher in ultrasound group than in shock wave group at 3 and 6 months after treatment (P<0.05). The AOFAS functional scores were increased in both groups (P<0.05) at 6 months after treatment, was significantly lower in ultrasound group than in shock wave group than group B (90.44∓13.27 vs 75.76∓21.40; P<0.05). The effective rates in shock wave group and ultrasound group were 92.31% and 76.32%, respectively (P<0.05). Recurrence was found in 1 patient (2.56%) in shock wave group and in 8 (21.05%) in ultrasound group (P<0.05). The patient satisfaction scores were significantly higher in shock wave group than in ultrasound group (8.13∓2.67 vs 6.63∓3.75, P=0.048).

CONCLUSIONPneumatic ballistic extracorporeal shock achieves better medium- and long-term outcomes than ultrasound-guided hormone injection in the treatment of plantar fasciitis.