Objective To investigate the correlations of serum matrix metalloproteinase-2, -9 (MMP-2, MMP-9) contents and tissue expressions in hepatocellular carcinoma with tumor invasion and metastasis. Methods Serum MMP-2, MMP-9 contents were detected in 40 patient with hepatocellular carcinoma and 20 healthy controls by ELISA; the expressions and distributions of MMP-2 and MMP-9 in 40 patients and 10 normal tissues were detected by immunohistochemical method. Results Serum MMP-2, MMP-9 contents were significantly elevated in cancer samples compared with normal serum (P

Objective To investigate the expressions of CD40 and ICAM-1 in primary hepatocarcinoma and clinical significance and analyze their relationships with carcinogenesis and progression of liver cancer. Methods S-P immunohistochemistry was used to detect the expressions of CD40 and ICAM-1 in 40 cases of hepatocarcinoma and 25 cases of normal liver tissue (distance from cancer tissues≥2 cm).Results The positive expression rates of CD40 and ICAM-1 were 47.5% and 82.5% in liver cancer tissue,which was significantly higher than those in normal liver tissue (P0.05).The positive expression rate of ICAM-1 was associated with lymph node metastasis as well as histological grade(P

AIM: To investigate effect of atrial natriuretic peptide (ANP) on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rat. METHODS: Mean arterial blood pressure (MAP) was recorded with model 6280 physiology intelligentialize grapher, nitric oxide (NO) and endothelin (ET) concentrations in plasma were measured after lipopolysaccharide (LPS) or following LPS ,ANP was injected into vein in rats. After experiment,lung water as well as pulmonary histopathological changes was measured and observed, respectively. RESULTS: Administration of LPS elicited a persistence decrease in MAP (8.1 kPa?2.6 kPa,at 4 h,P0.05); The histopathological of lung displayed markedly improved. CONCLUSION: ANP attenuates ALI induced by LPS in the rat. The effect of ANP may be via decreasing secretion of ET,NO and regulation arterial blood pressure. [

Objective To investigate the technical feasibility of medical robot in application to vascular intervention. Methods The independent-developed medical robot was used in the glass vessel model and vascular intervention experiments in a dog. Results The process of experiments were smooth,the system movement did not have any malfunction,and the animal experiments did not have any operative complications. The operative time was 50 minutes.Conclusions The medical robot can basically meet the requirements of cerebral angiography. It has laid a foundation for further development of intracranial vascular interventional procedures and clinical application.

Objective:To investigate whether endogenous hydrogen sulfide (H2 S)was involved in the pathogenesis of osteoarthritis (OA)and its underlying mechanism,to detect H2 S and its synthases ex-pression in knee cartilage in patients diagnosed with different severity of OA,and to explore the transcrip-tion and expression of gene MMP-13 in chondrocytes treated with IL-1βor H2S.Methods:Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2 S content using methylene blue assay.Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes.The chondrocytes were cultured to the P3 generation and H2 S molecular probes were used for detection of endogenous H2 S generation in the chondrocytes.Immunocytochemistry was used to detect the localization of H2 S synthases including cystathionine β-synthase (CBS),cystathionine-γ-lyase (CSE),and mercap-topyruvate sulfurtransferase (MPST)in OA chondrocytes.Western blot was used to quantify the protein expressions of CSE,MPST,and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty.The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments:(1 )the normal control group,no reagent was added;(2)the IL-1βgroup,5 μg/L of IL-1βwas added;(3)the IL-1β+H2S group,200 μmol/L of NaHS was added 30 min before adding 5 μg/L of IL-1β;(4)the H2 S group,200 μmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot,respectively.And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot.Results:The content of H2 S in the synovial fluid of degenerative knee was (14.3 ±3.3)μmol/L.Expressions of endogenous H2 S and its synthases including CBS,CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading)cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67 ±0.09 vs.1.26 ±0.11,P<0.05).However,no significant difference of CBS or MPST expression among the different groups was observed.The expression of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (1 .87 ±0.67 vs.0.22 ± 0.10,P<0.05 ),and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (0.55 ±0.11 vs.1.87 ±0.67,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The transcription of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (31.40 ±0.31 vs.1.00 ±0.00,P<0.05), and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (24.41 ± 1.28 vs.31.40 ±0.31,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The total NF-κB p65 in the IL-1βgroup was significantly higher than that in the normal chondrocytes (2.13 ±0.08 vs.0.73 ±0.08,P<0.05),and that in the IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (1 .24 ±0.13 vs.2.13 ±0.08,P<0.05 ),and that in the H2 S group had no significant difference compared with that in the normal control group.The phosphorylated NF-κB p65 in IL-1βgroup was significantly higher than that in the normal chondrocytes (1.30 ±0.13 vs.0.19 ±0.04,P<0.05),and that in IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (0.92 ±0.26 vs.1.30 ±0.13,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.Conclusion:H2 S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.

Objective To observe the effects of fecal microbiota transplantation on intestinal microbiota and brain function in sepsis rats. Methods Sixty male Sprague Dawley (SD) rats were divided into sham operation group, model group and fecal microbiota transplantation (FMT) group by random number table, each group 20 rats. The rat model of sepsis was established by injection of lipopolysaccharide (LPS) 10 mg/kg in tail vein. FMT group received nasogastric infusion of feces from healthy donor. Fecal samples were collected on the 6th day after the modeling to detect the levels of intestinal microbiota composition; the brain function was also evaluated by electroencephalogram (EEG), and the proportion of each waveform in EEG was calculated. After sacrifice of rats in different groups, the brain tissues were taken, the levels of protein expression and positive cells of Iba-1 in brain tissue were detected by Western Blot and immunohistochemistry method. Results ① Intestinal flora analysis showed that: the diversity index and Chaol index of the intestinal microbiota in model group were significantly lower than that in sham operation group (observed species:282±40 vs. 473±37, Chao1 index: 730±21 vs. 837±27, both P < 0.05); compared with the model group, the diversity index and Chaol index in FMT group were obviously higher (observed species: 461±20 vs. 282±40, Chao1 index:840±16 vs. 730±21, both P < 0.05). At phylum, family, genus level analysis showed that the proportion of Firmicutes phylum and Fusobacterium were obviously lower than those of sham operation group [Firmicutes phylum (22.12±1.34)% vs. (78.01±1.23)%, Fusobacterium: (2.03±0.17)% vs. (5.03±0.19)%, both P < 0.05], and the proportions of Proteobacteria, Bacteroidetes phyla and Acidaminococcaceae, Fusobacteriaceae, Enterbacteriacecae, Alistipes were markedly higher in model group [Proteobacteria: (70.21±2.35)% vs. (19.45±2.17)%, Bacteroidetes phyla: (4.12±0.19)% vs. (2.50±0.64)%, Acidaminococcaceae: (12.51±0.87)% vs. (1.01±0.12)%, Fusobacteriaceae: (13.62±1.27)% vs. (2.31±0.19)%, Enterbacteriacecae: (18.24±2.13)% vs. (4.15±1.51)%, Alistipes: (4.53±0.27)% vs. (1.47±0.33)%, all P < 0.05]; compared with the model group, the proportion of Firmicutes phylum and Faecalibacterium in FMT group were significantly higher [Firmicutes phylum: (72.14±2.31)% vs. (22.12±1.34)%, Faecalibacterium: (5.01±0.27)% vs. (2.03±0.17)%, both P < 0.05], and Proteobacteria, Bacteroidetes phyla and Acidaminococcaceae, Fusobacteriaceae, Enterbacteriacecae in FMT group were obviously lower [Proteobacteria: (14.23±1.98)% vs. (70.21±2.35)%, Bacteroidetes phyla: (3.15±0.18)% vs. (4.12±0.19)%, Acidaminococcaceae: (0.91±0.11)% vs. (12.51±0.87)%, Fusobacteriaceae: (1.25±0.15)% vs. (13.62±1.27)%, Enterbacteriacecae: (3.50±0.21)% vs. (18.24±2.13)%, all P < 0.05]. ② EEG analysis showed that the percentages of δ wave in EEG in model group was significantly higher after modeling than that in sham operation group [(16.86±0.50)% vs. (10.67±0.65)%, P < 0.05]; the ratios of δ wave in EEG was significantly lower in FMT group than that in the model group [(12.87±0.60)% vs. (17.35±0.41)%, P <0.05]. The incidence of abnormal EEG in sham operation group was 0, the incidence of abnormal EEG in model group was significantly increased [the ratios of δpredominant wave, θpredominant wave, low-voltage were 66.7% (6/9), 66.7% (6/9), 77.8% (7/9) respectively], the ratios of above abnormal waves in EEG in FMT group were obviously lower than those in model group [the ratios of above abnormal waves in FMT group were respectively 9.1% (1/11), 9.1% (1/11), 18.2%(2/11)]. ③ Western Blot analysis showed that the protein expression of Iba-1 in cortex in model group obviously was higher than that in sham operation group (Iba-1/β-actin: 1.39±0.16 vs. 0.67±0.18, P < 0.05); the expression of Iba-1 in cortex tissue of FMT group was markedly lower than that in model group (Iba-1/β-actin: 0.51±0.14 vs. 1.39±0.16, P < 0.05). ④ Immunohistochemistry of Iba-1 in cortex analysis showed that there were no Iba-1 positive cells in the cortex in sham operation group; Iba-1 positive cells were found in the cortex in model group; the number of Iba-1 positive cells in FMT group was less than that in model group. Conclusion FMT can improve the construction of intestinal microbiota, and ameliorate the brain dysfunction in SAE.

Objective To understand the infection status of enterovirus 71(EV71) and coxsackievirus A16(Cox A16) among children receiving health examination for child care setting entrance in Beijing and their related medical care seeking practice and provide evidence for the estimation of disease burden caused by hand foot and mouth disease(HFMD). Methods Serological survey was conducted in the local children receiving health examination for child care setting entrance. Enzyme-linked immunosorbent assay (ELISA) was conducted to detect anti-EV71 and anti-Cox A16 IgG and IgM. Results A total of 813 children were surveyed(mean age:3.5±1.0 year old). The seropositive rate was 61.9%and 4.4%for anti-Cox A16 IgG and IgM. The seropositive rate was 9.3% and 1.1% for anti-EV71 IgG and IgM. No significant difference was observed in sex specific seropositive rate (P>0.05). However,significant differences were found in seropositive rate among different age groups(P<0.05). Among the children who were anti-Cox A16 positive, 7.8%had ever had rashes on their hands and feet,mouth or buttocks(HFMD-like rashes). Among the children who were anti-EV71 positive,10.7%had ever had HFMD-like rashes. For the children who were anti-Cox A16 or anti-EV71 positive,only 7.1% were brought to see doctors by their parents. However,among the seropositive children with rashes,80.5% were brought to see doctors. Conclusion In the healthy children at the age to go to child care setting in Beijing,most had ever infected with Cox A16. The anti-EV71 positive rate was much lower than the anti-Cox A16 positive rate. It was necessary to strengthen the prevention and control of EV71 infection in child cares settings.

Objective: To explore the virulence related risk factors based on the enterovirus A71 (EV-A71) genome. Methods: The pairwise distance of each section of gene between mild and fatal cases was analyzed. The ⅴ domain of 5′UTR from mild and fatal cases in this study were constructed. Amino acid sequences of EV-A71 were analyzed to find the potential virulence regions which were statistically different between fatal and mild cases. Results: The two EV-A71 genome sequences in this study belonged to C4a genotype with the genomic homology of 96.2%-97.5%. The nucleotides in the ⅴ domains of the 5 ′UTR of EV-A71 from mild and fatal cases were the same. Each gene of EV-A71 from 31 mild cases and 30 fatal cases shared high homology. A total of four potential virulence sites (2 A: R68 M、2C: K41R、3 A: T/V47 A and 3C: I158 V) which were significantly different between mild cases and fatal cases were obtained. Conclusions The four sites in the unstructured protein coding region might be related with the virulence of EV-A71.

Objective To understand the distribution of virulence gene and the epidemiological characteristics of diarrheagenic Escherichia(E.) coli (DEC) from diarrheal patients in Beijing. Methods Stool specimens from diarrheal patients were cultured which were collected from the hospitals under sentinel surveillance program,during 2012-2013. DNA was examined by real-time PCR. Results 253 out of 6 370 specimens were positive for DEC detection with the rate as 4.0%. A total number of 262 DEC strains were isolated. Two different pathotypes of DEC strains with mixed infection,were isolated from 9 specimens. Different pathotypes would show the following profiles:42.8% for enteropathogenic E. coli (EPEC) including 42.0% atypical and 0.8% typical;38.9% for enterotoxigenic E. coli(ETEC)including 24.8%st positive,9.9%lt positive and 4.2%st and lt both positive;15.3% for enteroaggregative E. coli(EAEC);2.7% for enteroinvasive E. coli(EIEC);one strain STEC with serotype O26 ∶ K60. ETEC had obvious characteristics on age. All kinds of DEC were isolated throughout the year with seasonal fluctuation. Conclusion DEC isolates from diarrheal patients in Beijing were dominated by EPEC and ETEC,with atypical ones accounted for the majority of EPEC. One specimen was found under mixed infection. Pathotypes DEC were found to have different age and seasonal distributions.

Objective:To understand the epidemiological characteristics and clinical features of rotavirus-, norovirus-, adenovirus-and astrovirus-associ ated acute gastroenteritis in children under 5 years old in Beijing from Octorber, 2015 to March, 2017.Methods:In the intestinal clinic of 6 hospitals in 6 districts of Beijing, information and stool samples of the first 30 patients with acute gastroenteritis who are under the age of 5 years are collected monthly.Rotavirus, norovirus, adenovirus and astrovirus are identified by PCR.Descriptive epidemiological method was used to describe the epidemiological characteristics of diarrhea caused by rotavirus, norovirus, adenovirus and astrovirus in Beijing. One-way analysis of variance was used to analyze the Vesikari clinical severity score of of acute gastroenteritis caused by each virus. Unconditional logistic regression analysis was used to analysis the associated factors of clinical features.Results:Of the 2 052 samples, 709 (34.6%) were non-mixed infections: the positive rate of rotavirus, norovirus, adenovirus and astrovirus were 20.0%, 7.5%, 4.2% and 2.9%, respectively. A total of 135 cases (6.6%) were mixed infection. The mean and standard deviation of Vesikari clinical severity score was 8.0±3.1 for rotavirus associated acute gastroenteritis, which was significantly higher than norovirus (6.4±2.4, P<0.001), adenovirus (6.2±2.1, P<0.001) and astrovirus (6.1±2.0, P<0.001). The comparison of clinical features showed that compared with astrovirus, the children under 5 years old infected with rotavirus were more likely to have a diarrhea ≥5 days ( OR=3.334), have vomiting ≥3 times within one day ( OR=8.788), have vomiting≥1 day ( OR=3.963), have a Vesikari clinical severity score ≥11 severe cases ( OR=13.194). Norovirus infected cases were prone to have vomiting≥3 times in 1 day ( OR=5.710).Adenovirus infected cases were prone to have a diarrhea≥5 days ( OR=2.616). When using rotavirus as a reference, children under 5 years of age were less likely to develop fever≥38.4 ℃ after infection with norovirus ( OR=0.397) or adenovirus ( OR=0.280). Conclusions:The results of this study showed that the characteristics of acute gastroenteritis caused by different viruses are different. The clinical symptoms caused by rotavirus are more serious. Children under 24 months of age are at high risk of rotavirus infection. Effective preventive measures such as vaccination should be taken as soon as possible.