OBJECTIVES: To compare the application effect of microwave digestion - vacuum filtration - automated scanning electron microscopy (MD-VF-Auto SEM) method and plankton gene multiplex PCR system in the diagnosis of drowning. METHODS: Lung, liver and kidney tissue of 10 non-drowning cases and 50 drowning cases were prepared for further MD-VF-Auto SEM method analysis and plankton gene multiplex PCR system analysis. The positive detection rate of the two methods in each tissue was calculated. RESULTS: The positive rate of the MD-VF-Auto SEM method detecting diatoms in drowning cases was 100%, and few diatoms were detected in the liver and kidney tissues of 6 non-drowning cases. By using the plankton gene multiplex PCR system, the diatom positive rate of drowning cases was 84%, and all the non-drowning cases were negative. There were significant differences in the positive rate of the liver, kidney tissues between MD-VF-Auto SEM method and plankton gene multiplex PCR system (P<0.05), as well as the total positive rate of cases. However, no significant differences were found in the positive rates of lung tissues (P>0.05). CONCLUSIONS MD-VF-Auto SEM method is more sensitive than plankton gene multiplex PCR system in diatom test. But the plankton gene multiplex PCR system can also detect plankton other than diatoms. Combination of the two methods can provide a more reliable basis for the diagnosis of drowning.
Diatoms/genetics* ; Drowning/diagnosis* ; Humans ; Liver ; Lung ; Microscopy, Electron, Scanning ; Multiplex Polymerase Chain Reaction ; Plankton/genetics*

OBJECTIVE: To investigate the effect of silencing CD46 and desmoglein 2 (DSG2) in host A549 cells on the entry of human adenovirus type 3 (HAdV-3) and type 7 (HAdV-7) and host cell secretion of inflammatory cytokines. METHODS: RNA interference technique was use to silence the expression of CD46 or DSG2 in human epithelial alveolar A549 cells as the host cells of HAdV-3 or HAdV-7. The binding of the viruses with CD46 and DSG2 were observed with immunofluorescence staining at 0.5 and 1 h after viral infection. The viral load in the host cells was determined with qRT-PCR, and IL-8 secretion level was measured using ELISA. RESULTS: In infected A549 cells, immunofluorescent staining revealed colocalization of HAdV-3 and HAdV-37 with their receptors CD46 and DSG2 at 0.5 h and 2 h after infection, and the copy number of the viruses increased progressively after the infection in a time-dependent manner. In A549 cells with CD46 silencing, the virus titers were significantly lower at 2, 6, 12 and 24 h postinfection in comparison with the cells without gene silencing; the virus titers were also significantly decreased in the cells with DSG2 silencing. The secretion level of IL-8 increased significantly in A549 cells without siRNA transfection following infection with HAdV-3 and HAdV-7 (P < 0.0001), but decreased significantly in cells with CD46 and DSG2 silencing (P < 0.0001). CONCLUSION HAdV-3 and HAdV-7 enter host cells by binding to their receptors CD46 and DSG2, and virus titer and cytokines release increase with infection time. Silencing CD46 and DSG2 can inhibit virus entry and cytokine IL-8 production in host cells.
A549 Cells ; Adenoviruses, Human/metabolism* ; Desmoglein 2/metabolism* ; Humans ; Interleukin-8 ; Membrane Cofactor Protein/genetics* ; RNA, Small Interfering

Background/Aims: Esophagogastric junction adenocarci-noma (EJA) is a malignant tumor associated with high mor-bidity and has attracted increasing attention due to a rising incidence and low survival rate. Pathological biopsy is the gold standard for diagnosis, but noninvasive and effective tests are lacking, resulting in diagnoses at advanced stages.This study explored the diagnostic value of insulin-like growth factor binding protein 7 (IGFBP7) in EJA. Methods: A total of 120 EJA patients and 88 normal controls were recruited, and their serum levels of IGFBP7 were measured by enzymelinked immunosorbent assay. Receiver operating character-istic (ROC) curve analysis was used to assess the diagnostic value, and Pearson chi-square analysis was used to evaluate the correlation between IGFBP7 and clinical parameters. Ka-plan-Meier survival analysis was carried out to assess the ef-fect of IGFBP7 on overall survival (OS). Results: The levels of IGFBP7 were higher in both early- and late-stage EJA patients than in normal controls (p<0.001). The area under the ROC curve for EJA patients was 0.794 (95% confidence interval [CI], 0.733 to 0.854), with a cutoff value of 2.716 ng/mL, a sensitivity of 63.3% (95% CI, 54.0% to 71.8%) and a specific-ity of 90.9% (95% CI, 82.4% to 95.7%). For the diagnosis of early-stage EJA, the same cutoff value and specificity were obtained, but the sensitivity of IGFBP7 was 54.3% (95% CI, 36.9% to 70.8%). Patients with low IGFBP7 protein expres-sion had lower OS than those with high expression (p=0.034).The multivariate analysis showed that IGFBP7 is an inde-pendent prognostic factor for EJA (p=0.011). Conclusions Serum IGFBP7 acts as a potential diagnostic and prognostic marker for EJA.

A typical clinical case of taking Dictamni Cortex(Baixianpi) powder was analyzed to study liver damage caused by Dictamni Cortex. Liver damage was diagnosed according to the integrated evidence chain method recommended by the Guideline for Diagnosis and Treatment of Herb-Induced Liver Injury. By analyzing clinical history and biochemistry and imaging examinations, underlying diseases, such as viral hepatitis, autoimmune liver disease and alcoholic liver disease, were excluded. Through the investigation of medication history, we made it clear that the patient only took Dictamni Cortex powder during the period, and thus suspected that the liver injury was induced by Dictamni Cortex. Furthermore, the quality of the drug was tested, and the results showed it was consistent with the quality standard of Chinese Pharmacopoeia. DNA barcoding showed that the drug was 100% similar with Dictamnus dasycarpus. Moreover, exogenous harmful substances and chemical drug additions were tested, and the results showed that the content of heavy metal, pesticide residues and microbial toxin were consistent with the required standards, and no chemical drug additions were found in Agilent Fake TCM-Drugs database. In summary, we confirmed that the clinical case of drug-induced liver injury was induced by D. dasycarpus with the dose of 15 g•d⁻¹, which exceeded the prescribed amount of Chinese Pharmacopoeia. According to the Guideline for Diagnosis and Treatment of Herb-Induced Liver Injury, the case of drug-induced liver injury induced by D. dasycarpus was confirmed, which provided a direct and reliable evidence for the study of risk of liver injury induced by D. dasycarpus and its relevant preparations.

Objective To analyze the effect and safety of profile pegylated interferon α-2a (PEG-IFN-α-2a) combined with ribavarin on hepatitis C virus (HCV)-related cirrhosis patients after splenic embolization.Methods Sixty-four HCV patients who underwent splenic embolization were randomly divided into treatment group and control group (32 cases in each group).Control group was given subcutaneous injection of recombinant human interferon α-2b (rhIFN α-2b) every other day,and oral administration of ribavarin 300 mg three times a day;Treatment group did not only receive the treatment above,but also received subcutaneous injection pegylated interferon α-2a 180 μg once a week.Both groups were treated for 12 weeks.HCV RNA positive rate,blood routine parameters,serum alanine aminotransferase (ALT),aspartate aminotransferase (AST) levels,and adverse drug reactions were compared between the two groups.Results Before treatment,HCV RNA positive rate of the two groups were both 100％ (32/32 cases),after treatment,HCV RNA positive rate of treatment group was 53.13％ (17/32 cases) and that of and the control group was 78.12％ (25/32 cases),the difference was statistically significant (P ＜ 0.05).Mter treatment,the ANC of treatment group was (4.76 ± 0.33) × 109/L and that of the control group was (3.92 ±0.17) × 109/L,PLT levels of treatment group was (142.73 ± 17.04) × 109/L and that of and the control group was (128.72 ± 17.96) × 109/L,WBC levels of treatment group was (8.15 ± 1.84) × 109/L and that of and the control group was (6.57 ± 1.69 × 10)9/L,and the difference was statistically significant (P ＜ 0.05).After treatment,the ALT of the experimental group was 41.2 ± 22.4U · L-1 and that of and the control group was (60.1 ± 21.9) U · L-1,AST levels of the experimental group was 33.67 ± 18.46 U · L-1 and that of and the control group was (51.22 ± 16.95) U · L-1;The AST/ALT values were 0.78 ± 0.69,1.27 ± 0.73 in treatment group and in the control group,and the difference was statistically significant (P ＜ 0.05).Adverse drug reactions in treatment group was 40.63％ (13/32 cases),while it was 28.12％ (9/32 cases) in control group,thedifference was not statistically significant (P ＞ 0.05).Conclusion PEG-IFN-α-2a combined with riba virin could not only reduce HCV RNA positive rates,but also improve the serum AST and ALT levels in the treatment of hepatitis C virus-related cirrhosis underwent splenic embolization,which is safe.

OBJECTIVEChinese allergic subjects have high levels of sensitization to house dust mite (HDM) and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households.

METHODSDust samples were collected from nine cities. Major allergens Der p 1 and Der f 1 from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.

RESULTSHDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from <0.006 to 9.2 µg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.

CONCLUSIONHDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.

Air Pollution, Indoor ; statistics & numerical data ; Allergens ; chemistry ; Animals ; Bedding and Linens ; China ; Cockroaches ; Dust ; analysis ; Housing ; Pyroglyphidae ; Seasons

OBJECTIVEBy testing the changes of telomere binding protein in malignant transformation BEAS-2B cells induced by coal tar pitch smoke extracts, to study the role of protection of telomeres 1 (POT1), telomeric repeat binding factor 1 (TRF1) and TRF2 in tumorgenesis that contact with coal tar pitch.

METHODSThe BEAS-2B cells were induced by coal tar pitch smoke extracts to form malignant transformation cell model in vitro. The gene expression levels of mRNA were assessed by real-time quantitative RT-PCR, the protein expression variations were determined by cell culture overslip of immunohistochemical methods.

RESULTSIn malignant transformation cells, the mRNA expression level (POT1: 0.63 ± 0.04, TRF1: 0.36 ± 0.01) and the protein expression level (POT1: 0.36 ± 0.05, TRF1: 0.09 ± 0.03) of POT1 and TRF1 was statistically significant decreased compared to that of BEAS-2B group (mRNA: POT1: 1.00 ± 0.04, TRF1: 1.01 ± 0.16; protein: POT1: 0.55 ± 0.07, TRF1: 0.27 ± 0.07) and DMSO group (mRNA: POT1: 0.89 ± 0.12, TRF1: 0.90 ± 0.08; protein: POT1: 0.55 ± 0.10, TRF1: 0.26 ± 0.04) (P < 0.05); mRNA expression level (1.45 ± 0.07) and the protein expression level (0.88 ± 0.06) of TRF2 was increased compared to that of BEAS-2B group (mRNA: 1.00 ± 0.07, protein: 0.48 ± 0.06) and DMSO group (mRNA: 1.00 ± 0.06, protein: 0.50 ± 0.06) (P < 0.05).

CONCLUSIONThe change of gene and protein expression level in POT1, TRF1, and TRF2 involved in the process that evolved into malignant transformation in bronchial epithelial cells BEAS-2B induced by coal tar pitch smoke extracts.

Cell Line ; Cell Transformation, Neoplastic ; metabolism ; Coal Tar ; toxicity ; Epithelial Cells ; cytology ; metabolism ; pathology ; Humans ; Repetitive Sequences, Nucleic Acid ; Telomere-Binding Proteins ; genetics ; metabolism

The primers and probes for the Real-time RT-PCR were designed based on the multiple sequence (swine and humans HEV strains) alignments of the ORF3 region of genotype 4 HEV. A rapid, sensitive and stable TaqMan Real-time RT-PCR assay was established, and its specificity and sensitivity were assessed, and comparison of the Real-time RT-PCR with conventional and nested RT-PCR was performed. The results found that the crossing points showed linearly proportional to the logarithm of the input copy number. The correlation coefficient (R2) and the slope value of the standard curves with plasmid DNA were 0.994 and -3.312, respectively. The efficiency (E) of the PCR was 100%. Coefficients of variation values of the different diluted plasmid DNA were low in the same or different repeated experimental group. In addition, the assay was able to correctly detect genotype 4 HEV RNA from swine fecal samples. The sensitivity of established assay was 100-fold higher than that of conventional RT-PCR and 10-fold higher than nested RT-PCR.
Animals ; DNA Primers ; genetics ; Disease Reservoirs ; virology ; Feces ; virology ; Fluorescence ; Genotype ; Hepatitis E ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Swine ; virology

OBJECTIVETo investigate the pathological feature of primary hepatic carcinoma and the clinical significance.

METHODSFrom August 2000 to December 2007, there were 89 patients with cirrhosis and carcinoma of liver who accepted whole liver resection. The whole liver was cut into 10 mm slices to examine the tumor size, number, distribution, capsule, satellite nodes, portal vein tumor thrombi (PVTT). The invaded adjacent tissue and lymph nodes were recorded, the distance from satellite to major tumor was measured, then histological examinations were carried out, and the final diagnosis was made by pathologists.

RESULTSThe total of 89 cases included hepatocellular carcinoma in 86 cases and cholangiocarcinoma in 3 cases; 53 cases with multiple tumors and 36 cases with solitary tumor; complete capsule only in 14 cases, no obvious margin in 11 cases, 13 cases had a major tumor in the right lobe and a small tumor in the left lobe; 8 of 25 cases with gross invaded tissue were confirmed by histological examination, 7 of 16 cases with swollen lymph nodes were infiltrated by cancer cells. There were 47 cases with PVTT (47.2%) and 39 cases with satellite nodes (43.8%). PVTT and satellite nodes increased with the increase of sizes and the numbers of the tumors. The distance from satellite node to major tumor mostly were 0.5 - 3.0 cm.

CONCLUSIONSThe whole explanted liver can completely reflect the characteristics of growth and infiltration of hepatic carcinoma. Attention must be paid to the small cancer lesions in another lobe, distal satellite nodes from major tumor, and tumor thrombi in a small branch of portal vein, which can not be found by imaging, and might influence the curative effectiveness after liver resection or transplantation.

Adult ; Carcinoma, Hepatocellular ; pathology ; surgery ; Female ; Hepatectomy ; Humans ; Liver ; pathology ; Liver Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Young Adult