[The Effect of TCP1 Expression on the Proliferation and the Accumulation of Intracellular Drug of HL60/A and HL60 Cell and Its Mechanism].

OBJECTIVE: To investigate the effect of TCP1 expression on the proliferation and the accumulation of intracellular drug of HL60/A and HL60 cells and its possible molecular mechanism. METHODS: Lentiviral transfection technology was used to construct HL60/A and HL60 cells with knocked down or overexpressed TCP1 and their control cells. The efficiency of knockdown and overexpression was evaluated by Western blot. The cell proliferation was detected by CCK-8 assay. The intracellular drug accumulation was detected by laser confocal detection and flow cytometry. The expression levels of MRP1, P-gP and p-AKT were evaluated by flow cytometry and Western blot. RESULTS: After TCP1 was knocked down,the proliferation ability of HL60/A cells was significantly reduced, the accumulation of intracellular drug was significantly increased and the expression of MRP1 and P-gP protein were decreased. After TCP1 was overexpressed, the proliferation ability of HL60 was significantly increased, the accumulation of intracellular drug was significantly decreased and the expression of MRP1 and P-gP protein were increased. Intervention of LY294002 significantly antagonized the promotion on cell proliferation, the inhibition on intracellular drug accumulation and the expression of MRP1 and P-gP mediated by TCP1 overexpressing in HL60 cells. CONCLUSION: TCP1 can promote cell proliferation, improve the expression of MRP1 and P-gP by activating PI3K/AKT signal, and reduce intracellular drug accumulation.

Correlation Between Substantia Nigra Hyperechogenicity and Iron Metabolism in the Postural Instability Gait Difficulty Subtype of Parkinson's Disease.

OBJECTIVE: The correlation between substantia nigra (SN) hyperechogenicity on transcranial sonography (TCS) and serum iron metabolism parameters in patients with the postural instability gait difficulty (PIGD) subtype of Parkinson's disease (PD) was investigated so as to explore the pathological mechanism of SN hyperechogenicity. METHODS: The study enrolled 95 PIGD patients recruited by the Parkinson's Disease Specialty in the Second Affiliated Hospital of Soochow University during June 2019-2021. On the basis of the TCS results, the PIGD patients were assigned to the PD with SN hyperechogenicity (SN+) group (n = 60) and PD without SN hyperechogenicity (SN-) group (n = 35). Meanwhile, 49 sex- and age-matched healthy individuals were included in the control group. All participants underwent blood tests. Differences in the iron metabolism parameters among the three groups and the correlation between SN hyperechogenicity and serum iron metabolism parameters were analyzed. RESULTS: Serum ferritin, ceruloplasmin and transferrin levels were lower in the SN+ and SN- groups than in the control group (all p values <0.001). The serum ceruloplasmin level was lower in the SN+ group (0.23 [0.20, 0.25] g/L) than in the SN- group (0.25 [0.22, 0.29] g/L) (p = 0.001), and the proportion of patients with an abnormal ceruloplasmin level was higher in the SN+ group than in the SN- group (43.3% [26/60] vs. 14.3% [5/35], χ2 = 8.484, p = 0.004). Moreover, the SN hyperechogenicity area was negatively correlated with the serum transferrin level (r = -0.428, p < 0.001). CONCLUSION: Decreased serum ceruloplasmin levels may be associated with SN hyperechogenicity development in PIGD patients. The SN hyperechogenicity area is negatively correlated with the serum transferrin level.

PRM-based quantitative proteomics analysis of altered HSP abundance in villi and decidua of patients with early missed abortion.

OBJECTIVE: In this study, we aimed to identify differentially expressed heat shock protein (HSP) profiles in the villi and decidua from patients with early missed abortion (EMA). METHODS: By using high-throughput and high-precision parallel reaction monitoring (PRM)-based targeted proteomics techniques, this study examined the abundance of HSPs in the villi and decidua of 10 patients with EMA and 10 controls. Moreover, the abundance of 3 HSPs in the villi of another 22 patients with EMA and 22 controls was verified with Western blotting and immunohistochemistry (IHC). RESULTS: There were potential differences in the abundance of 16 HSPs and 42 polypeptides in human villi and decidua compared with those of the control group. Among them, HSP90AB1, HSPD1 and HSPA13 were downregulated in abundance in villi of patients with EMA, with a statistically significant difference, which was consistent with the verification results of Western blots and IHC. CONCLUSION: Using a PRM-based targeted proteomics technique, this study is the first to screen and quantitatively analyze the expression profile of HSPs in the villi and decidua of patients with EMA. The significant downregulation of HSP90AB1, HSPD1 and HSPA13 was found to have a potentially intimate association with the occurrence of EMA. The findings in our study may provide novel potential research targets related to HSPs for the pathogenesis, prevention and treatment of EMA.

Diagnostic Value of Procalcitonin and Interleukin-6 on Early Postoperative Pneumonia after Adult Cardiac Surgery: A Prospective Observational Study.

BACKGROUND: Postoperative pneumonia (PP) is the most common primary infection after cardiac surgery, increasing the hospitalization expense and causing the consumption of healthcare resources. This study aimed to investigate the diagnostic value of procalcitonin (PCT) and interleukin-6 (IL-6) on early postoperative pneumonia after adult cardiac surgery. METHODS: In this prospective observational study, patients with pneumonia and age- and sex-matched cases in our center from October 10, 2020 to January 31, 2021 were included. Patients diagnosed with pneumonia in this study needed meet both clinical and microbiological diagnostic criteria. Blood samples were collected in all patients from postoperative day (POD) 1 to postoperative day 5 to detect PCT, IL-6, white blood cell count, and C-reactive protein. The diagnostic performance of different biomarkers was evaluated by the receiver operating characteristic curves and the area under the curves. RESULTS: Our study enrolled 272 patients, including 24 patients with postoperative pneumonia and 248 age- and sex-matched cases. From POD1 to POD5, the absolute value of PCT and PCT variations showed diagnostic significance for pneumonia (P < .05); the diagnostic value of the absolute value of IL-6 and IL-6 variations was not satisfying. White blood cell count showed no differences; C-reactive protein had no diagnostic value before POD4. Multivariable logistic regression showed that PCT variation and IL-6 variation from POD3 to POD1 were the strongest risk factors for postoperative pneumonia [OR:12.50, 95% CI: (3.40-45.5); OR:13.71, 95% CI: (1.11-168.47)]. According to the above results, we defined the PL Index. PL Index showed the best diagnostic value among those biomarkers in POD3 [AUC: 0.90, 95% CI: (0.79-0.95)]. Multivariable logistic regression showed PL Index POD3 has significant correlation with postoperative pneumonia [OR:1.23, 95% CI: (1.11-1.37), P = .041]. CONCLUSIONS: PCT variation and IL-6 were more accurate than C-reactive protein and white blood cell count to predict early postoperative pneumonia, but the diagnostic properties of PCT could not be observed during the first three postoperative days due to the inflammatory process. By combining the variations of PCT and IL-6, we defined the PL Index, which shows the best diagnostic ability on early postoperative pneumonia after adult cardiac surgery.

Mitochondria autophagy: a potential target for cancer therapy.

Mitophagy is a selective form of macroautophagy in which dysfunctional and damaged mitochondria can be efficiently degraded, removed and recycled through autophagy. Selective removal of damaged or fragmented mitochondria is critical to the functional integrity of the entire mitochondrial network and cells. In past decades, numerous studies have shown that mitophagy is involved in various diseases; however, since the dual role of mitophagy in tumour development, mitophagy role in tumour is controversial, and further elucidation is needed. That is, although mitophagy has been demonstrated to contribute to carcinogenesis, cell migration, ferroptosis inhibition, cancer stemness maintenance, tumour immune escape, drug resistance, etc. during cancer progression, many research also shows that to promote cancer cell death, mitophagy can be induced physiologically or pharmacologically to maintain normal cellular metabolism and prevent cell stress responses and genome damage by diminishing mitochondrial damage, thus suppressing tumour development accompanying these changes. Signalling pathway-specific molecular mechanisms are currently of great biological significance in the identification of potential therapeutic targets. Here, we review recent progress of molecular pathways mediating mitophagy including both canonical pathways (Parkin/PINK1- and FUNDC1-mediated mitophagy) and noncanonical pathways (FKBP8-, Nrf2-, and DRP1-mediated mitophagy); and the regulation of these pathways, and abovementioned pro-cancer and pro-death roles of mitophagy. Finally, we summarise the role of mitophagy in cancer therapy. Mitophagy can potentially be acted as the target for cancer therapy by promotion or inhibition.

Relationship of programmed death ligand-1 expression with clinicopathological features and prognosis in patients with oral squamous cell carcinoma: A meta-analysis.

BACKGROUND: Programmed death ligand-1 (PD-L1) expression is related to the prognosis of many solid tumors; however, its prognostic value in oral squamous cell carcinoma (OSCC) remains unclear. Here, a meta-analysis was performed to estimate the association of PD-L1 expression with prognosis and clinicopathological features in patients with OSCC. METHODS: PubMed, Web of Science, EMBASE, Cochrane Library, and CNKI databases were searched to find relevant studies for identification of the association of PD-L1 expression with clinicopathological features and overall survival (OS) in patients with OSCC. The strength of the association of PD-L1 expression with clinicopathological features and OS in patients with OSCC was assessed according to the relative risk (RR), hazard ratio (HR), and 95% confidence interval CI (CI). RESULT: Twenty-three studies (including 3217 patients with OSCC) were evaluated. The meta-analysis showed that positive PD-L1 expression was significantly correlated with OS in patients with OSCC (HR = 1.00, 95% CI = 0.76-1.30, p = 0.284). Positive PD-L1 expression was significantly correlated with sex (RR = 1.22, 95% CI = 1.07-1.38, p = 0.002), histological differentiation (RR = 1.15, 95% CI = 1.02-1.30, p = 0.020), distant metastasis (RR = 0.68, 95% CI = 0.54-0.86, p = 0.011), lymph node metastasis status (RR = 0.83, 95% CI = 0.76-0.91, p < 0.001), TNM stage (RR = 0.81, 95% CI = 0.73-0.89, p < 0.001), and human papilloma virus infection status (RR = 1.30, 95% CI = 1.04-1.62, p = 0.019), but was not correlated with T stage and tumor recurrence. CONCLUSION: High PD-L1 expression in OSCC was not related to OS. However, high PD-L1 expression was significantly related to certain clinicopathological features. Thus, positive PD-L1 expression may be a biomarker of poor prognosis in patients with OSCC.

Association of serum adiponectin level with cystatin C in male patients with obstructive sleep apnea syndrome.

PURPOSE: Obstructive sleep apnea syndrome (OSAS) was suggested to exert an effect on renal function. However, the specific mechanism was still unknown. We try to find the association among OSAS, adiponectin, and cystatin C and the effect of adiponectin on renal function in OSAS patients. METHODS: Seventeen healthy men and seventy-three men which only had OSAS were included in the end. Apnea-hypopnea index (AHI), oxygen desaturation index (ODI), the percentage of total sleep time spent with SpO2 < 90% (T90%), lowest O2 saturation (LaSO2), Epworth Sleepiness Scale (ESS) score, serum adiponectin, and high-sensitive C-reactive protein (hsCRP) were detected in all subjects, and renal function was evaluated with creatinine, cystatin C, and estimated glomerular filtration rate (eGFR). RESULTS: Demographic data, creatinine, and eGFR did not differ among the studied groups. Decreased serum adiponectin levels were associated with severe OSAS. OSAS patients had a higher hsCRP and cystatin C than those without OSAS. Serum adiponectin levels had a negative association with cystatin C. After adjusted for confounders, adiponectin, hsCRP, and ODI had a significant prediction on the cystatin C (ß = - 0.218, p = 0.011; ß = 0.226, p = 0.037; and ß = 0.231, p = 0.029). CONCLUSIONS: Decreased serum adiponectin was associated with increased cystatin C in male OSAS patients. These results suggest that serum adiponectin might be a regulatory factor for renal function in OSAS.

Deoxynivalenol Induces Intestinal Damage and Inflammatory Response through the Nuclear Factor-κB Signaling Pathway in Piglets.

Deoxynivalenol (DON) is highly toxic to animals and humans, but pigs are most sensitive to it. The porcine mucosal injury related mechanism of DON is not yet fully clarified. Here, we investigated DON-induced injury in the intestinal tissues of piglet. Thirty weanling piglets [(Duroc × Landrace) × Yorkshire] were randomly divided into three groups according to single factor experimental design (10 piglets each group). Piglets were fed a basal diet in the control group, while low and high dose groups were fed a DON diet (1300 and 2200 µg/kg, respectively) for 60 days. Scanning electron microscopy results indicated that the ultrastructure of intestinal epithelial cells in the DON-treated group was damaged. The distribution and optical density (OD) values of zonula occludens 1 (ZO-1) protein in the intestinal tissues of DON-treated groups were decreased. At higher DON dosage, interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α mRNA levels were elevated in the intestinal tissues. The mRNA and protein levels of NF-κB p65, IκB-α, IKKα/ß, iNOS, and COX-2 in the small intestinal mucosa were abnormally altered with an increase in DON concentration. These results indicate that DON can persuade intestinal damage and inflammatory responses in piglets via the nuclear factor-κB signaling pathway.

Androgen alleviates neurotoxicity of ß-amyloid peptide (Aß) by promoting microglial clearance of Aß and inhibiting microglial inflammatory response to Aß.

AIMS: Lower androgen level in elderly men is a risk factor of Alzheimer's disease (AD). It has been reported that androgen reduces amyloid peptides (Aß) production and increases Aß degradation by neurons. Activated microglia are involved in AD by either clearing Aß deposits through uptake of Aß or releasing cytotoxic substances and pro-inflammatory cytokines. Here, we investigated the effect of androgen on Aß uptake and clearance and Aß-induced inflammatory response in microglia, on neuronal death induced by Aß-activated microglia, and explored underlying mechanisms. METHODS: Intracellular and extracellular Aß were examined by immunofluorescence staining and Western blot. Amyloid peptides (Aß) receptors, Aß degrading enzymes, and pro-inflammatory cytokines were detected by RT-PCR, real-time PCR, and ELISA. Phosphorylation of MAP kinases and NF-κB was examined by Western blot. RESULTS: We found that physiological concentrations of androgen enhanced Aß42 uptake and clearance, suppressed Aß42 -induced IL-1ß and TNFα expression by murine microglia cell line N9 and primary microglia, and alleviated neuronal death induced by Aß42 -activated microglia. Androgen administration also reduced Aß42 -induced IL-1ß expression and neuronal death in murine hippocampus. Mechanistic studies revealed that androgen promoted microglia to phagocytose and degrade Aß42 through upregulating formyl peptide receptor 2 and endothelin-converting enzyme 1c expression, and inhibited Aß42 -induced pro-inflammatory cytokines expression via suppressing MAPK p38 and NF-κB activation by Aß42 , in an androgen receptor independent manner. CONCLUSION: Our study demonstrates that androgen promotes microglia to phagocytose and clear Aß42 and inhibits Aß42 -induced inflammatory response, which may play an important role in reducing the neurotoxicity of Aß.

Research advances on animal genetics in China in 2015.

Chinese scientists have made significant achievements in the field of animal genetics in 2015. Incomplete statistics show that among all the publications of 2015 involving nematode (Caenorhabditis elegans), fly (Drosophila melanogaster), zebrafish (Danio rerio), African clawed frog (Xenopus) or mice (Mus musculus), about 1/5 publications are from China. Many innovative studies were published in high-impact international academic journals by Chinese scientists, including the identification of a putative magnetic receptor MagR, the genetic basis for the regulation of wing polyphenism in the insect brown planthopper (Nilaparvata lugens), DNA N6-methyladenine (6mA) modification in the Drosophila genome, a novel molecular mechanism regarding the dendritic spine pruning and maturation in the mammals, the mechanism for the CREB coactivator CRTC2 in the regulation of hepatic lipid metabolism, the control of systemic inflammation by neurotransmitter dopamine, the role of Gasdermin protein family in triggering pyroptosis, a parvalbumin-positive excitatory visual pathway to trigger fear responses in mice, etc. Chinese scientists have also made important contributions in genome editing via TALEN or CRISPR/Cas system. According to incomplete statistics, more than 1/5 of the publications related to genome editing in 2015 are from China, where a variety of animals with different approaches were targeted, ranging from the worm to primates. Particularly, CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes was successfully achieved for the first time. China has been one of the leading countries in genome sequencing in recent years, and Chinese scientists reported the sequence and annotation of the genomes of several important animal species in 2015, including goose (Anser cygnoides), Schlegel's Japanese Gecko (Gekko japonicus), grass carp (Ctenopharyngodon idellus), large yellow croaker (Larimichthys crocea) and pig (Sus scrofa). They further analyzed the genome-wide genetic basis of the species-specific physiological and pathological characteristics as well as their adaptation to environmental conditions. In this review, we make a first attempt to summarize the research advances on animal genetics in China in 2015, with an emphasis on the achievements led by Chinese scientists and carried out in Chinese institutions. We will briefly discuss the significance of their research and contributions of Chinese scientists in animal genetics.

MicroRNA-744 inhibited cervical cancer growth and progression through apoptosis induction by regulating Bcl-2.

Growing evidence suggests that microRNA plays an essential role in the development and metastasis of many tumor progressions, including cervical cancer. Aberrant miR-744 expression has been indicated in many growth of tumor, the mechanism of miR-744 inhibits both the proliferation and metastatic ability for cervical cancer remains unclear. Accumulating evidences reported that Bcl-2 signal pathway plays an important role in the cellular process, such as apoptosis, cell growth and proliferation. The goal of this study was to identify miR-744 that could inhibit the growth, migration, invasion, proliferation and metastasis of gastric cancer through targeting Bcl-2 expression. Real-time PCR (RT-qPCR) was used to quantify miR-744 expression in vitro and vivo experiments. The biological functions of miR-744 were determined via cell proliferation. Our study indicated that miR-744 targeted on Bcl-2, which leads to the inactivation of apoptosis signaling and the cell proliferation of cervical cancer cells, ameliorating cervical cancer growth and progression. In addition, both up-regulation of miR-744 and down-regulation of Bcl-2 could stimulate Caspase-3 expression, promoting apoptosis of cervical cancer cells. Therefore, our research revealed the mechanistic links between miR-744 and Bcl-2 in the pathogenesis of cervical cancer through modulation of Caspase-3, leading to the inhibition of cervical cancer cell growth. And targeting miR-744 could be served as a novel strategy for future cervical cancer therapy clinically.

Complete mitochondrial genome sequence and mutations of the laryngeal cancer model inbred nude rat strain.

In the present work, we undertook the complete mitochondrial genome sequencing of an important laryngeal cancer model inbred rat strain for the first time. The total length of the mitogenome was 16,308 bp. It harbored 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and one non-coding control region (D-loop region). The mutation events were also reported.

[Expression and significance of matriptase in ovarian cancer cells with diverse metastatic potential].

OBJECTIVE: To study the expression and significance of matriptase in different metastatic potential of human ovarian cancer cells. METHODS: High-metastatic human ovarian cancer cell HO8910PM and ovarian cancer cell HO8910 were collected.The ability of metastatic of the former was stronger than that of the latter. Compared the ability of invasion and migration in HO8910PM and HO8910 by scratch assay and by millicell chamber artificial reconstituted basement membrane invasion assay. Detected the matriptase mRNA and protein expression levels in HO8910PM and HO8910 through reverse transcription(RT)-PCR and immunocytochemistry methods. RESULTS: The 24 hours' migration distance (347 ± 8) µm of HO8910PM cells were significantly higher than that in HO8910 group (154 ± 10) µm (P < 0.01);The number of HO8910PM cells that penetrated the matrigel after 24 hours' incubation were significantly higher than that in HO8910 group (90.7 ± 2.1 vs 63.3 ± 1.5,P < 0.01). The expression of matriptase mRNA in HO8910PM cells was higher than that in HO8910 group (0.72 ± 0.03 vs 0.38 ± 0.04,P < 0.01). The migration was positively correlated with the matriptase mRNA expression levels (r = 0.992, P < 0.01); and the invasiveness was also positively correlated with the matriptase mRNA expression levels (r = 0.973, P < 0.01). As far protein level,the expression of matriptase protein in HO8910PM cells was higher than that in HO8910 group (15.6 ± 0.8 vs 7.6 ± 1.3,P < 0.01). The migration was positively correlated with matriptase protein expression levels (r = 0.971, P < 0.01); And the invasiveness was also positively correlated with the matriptase protein expression levels (r = 0.958, P < 0.01). CONCLUSIONS: The relationship between the expression levels of matriptase and the metastatic of ovarian cancer cells may be correlative. The function of matriptase in ovarian cancer cells metastatic mechanism still need to be confirmed.

[Differentiation of human promyelocytic leukemia HL-60 cells induced by proanthocyanidin and its mechanism].

This study was purposed to investigate the proliferation, differentiation and apoptosis of human promyelocytic leukemia HL-60 cells induced by proanthocyanidin (PAC). HL-60 cells were incubated with 20 mg/L PAC for 24 h, the cell growth was evaluated by CCK-8 assay. the effect of PAC on HL-60 cells was evaluated and the cells morphology was observed by optical microscopy. Expression of CD14 and CD11b, and cell cycle were analyzed by flow cytometry. The results showed that the growth of HL-60 cells was inhibited after treatment with PAC of different concentration in a dose-dependent manner (P < 0.05). 20 mg/L PAC displayed significant effect on HL-60 cells with inhibition ratio (72.3 ± 1.8)% for 24 h. Microscopy displayed that some cells differentiated to relative mature cells after treating for 48 h. Expression of CD14 increased and the expression of CD11b increased a little after treating with 20 mg/L PAC for 24 h, the ratio of cells in G0/G1 phase increased, but the ratio of cells in S phase decreased. The mRNA and protein expression of P21 gene increased, but the protein expression of CDK4 and Cyclin D1 decreased. It is concluded that PAC may inhibit the proliferation of HL-60 cells in vitro, induces the differentiation of HL-60 cells, and arrests the cells in G0/G1 phase. The possible mechanism may be related to up-regulation of P21 gene expression and down-regulation of the protein expression of CDK4 and Cyclin D1.

Substituted benzamides containing azaspiro rings as upregulators of apolipoprotein A-I transcription.

Apolipoprotein A-I (Apo A-I) is the principal protein component of high density lipoprotein (HDL), which is generally considered as a potential therapeutic target against atherosclerosis. The understanding of the Apo A-I regulation mechanism has fuelled the development of novel HDL targeted therapeutic approaches. To identify novel agents that can upregulate Apo A-I expression, we performed a cell-based reporter assay to screen 25,600 small molecules. Based on the dataset obtained from screening, a series of novel analogs of substituted benzamides containing azaspiro rings were assessed for their ability to induce the transcription of the Apo A-I gene, and the structure-activity relationship (SAR) around these analogs was also proposed. The results indicated that the trifluoromethyl substituted benzamide containing an azaspiro ring is a promising backbone for designing Apo A-I transcriptional upregulator and could be viable leads for development of new drugs to prevent and treat atherosclerosis in the future.

[Chemopreventive effect of boswellic acid and curcumin on 7,12-dimethyl benzanthracene-induced hamster cheek pouch carcinogenesis].

OBJECTIVE: To evaluate the chemopreventive effects of boswellic acid and curcumin on 7,12-dimethyl benzanthracene(DMBA)-induced oral carcinogenesis in the hamster cheek pouch model. METHODS: Male Syrian golden hamsters (6 - 8 weeks old, 80 - 130 g in weight) were randomly divided into seven groups, with group A serving as the untreated negative control. The left cheek pouch of the remaining hamsters was topically treated with 0.5% DMBA in mineral oil three times a week for 6 weeks. They were then randomized to six groups with group B serving as a positive control and receiving no further treatment. Groups C-G were treated topically with 5, 10 mg/L boswellic acid, 5, 10 µmol/L curcumin, or the combination of 5 mg/L boswellic acid and 5 µmol/L curcumin three times per week for 18 weeks. The animals were injected with bromodeoxyuridine intraperitoneally at 50 mg/kg 2 h prior to killing. At the 25 th week all the hamsters were sacrificed and cheek pouch tissue was harvested. One half of the tissue was snap frozen in liquid nitrogen for analysis of arachidonic acid metabolites, and the other half was fixed in 10% phosphate-buffered saline(PBS)-buffered formalin for histopathological examination. RESULTS: Six-weeks of DMBA followed by 18-weeks of topical application of boswellic acid and curcumin, both boswellic acid (5, 10 mg/L) and curcumin (5, 10 µmol/L) significantly inhibited the incidence from 93.8% to 73.9% (P > 0.05), numbers from 2.19 ± 0.98 to 1.13 ± 0.81 (P < 0.01) and size of visible tumors. Microscopically the incidence of squamous cell carcinoma and BrdU index were also significantly suppressed by boswellic acid and curcumin. CONCLUSIONS: Both boswellic acid and curcumin were effective in preventing oral carcinogenesis in DMBA-induced hamster cheek pouch model.

[Preparation of recombinant polypeptide of N-terminal heparin-binding domain of fibronectin and its effect on disseminated intravascular coagulation in rats].

This study was aimed to prepare the polypeptide of N-terminal heparin-binding domain of fibronectin(rhFNHN-29 polypeptide) with pichia expression system, to detect biological activity of recombinant polypeptide and investigate its effect on disseminated intravascular coagulation (DIC) in rats. The sequence of N-terminal heparin-binding domain of fibronectin was amplified from FNcDNA by PCR. The aim gene was cloned into T vector for selection. Then it was cloned into pAo815SM and pPIC9K vectors.Lined pPIC9K vectors were transformed into GS115 Pichia cells so as to express the aim polypeptide in Pichia expression system. The fermentation liquid were precipitated by 80% ammonium sulfate, and the further dissolved sediment were purified using S-100 column and SP column. Its activity of binding with heparin were detected by Western-blot. The established DIC rats (40 rats) were randomly divided into two groups. One group was treated with rhFNHN-29 polypeptide, and the other was treated with normal saline. The rats in the former group were injected with rhFNHN-29 polypeptide (10 mg/kg) through tail vein at 0.5 hour before, 2 hours and 4 hours after injection of LPS respectively. The rats in latter group were injected with equal volume saline. In addition, 20 normal rats injected with normal saline were as normal controls. 500 microl blood was taken from the rat vein, at 6 hours after the injection of LPS. White blood cell (WBC), hemoglobin (Hb) and platelets were tested from 50 microl blood. The rest 450 microl blood was used to isolate plasma for detecting TNFa level and coagulogram. The rats were killed at 24 hours after injection with LPS. Their livers, lungs, hearts, kidneys, and brain tissues were taken for histopathologic examination. The results showed that the aim polypeptide was successfully expressed in Pichia expression system. The expression level reached approximately 30 mg/L. The polypeptide had activity of binding with heparin antibody. In the experiment study of polypeptide effect on DIC in rats, the plasma TNFa level in polypeptide-treated group was lower than that in saline control group, the hemogram, coagulogram and histopathology were more obviously improved in polypeptide-treated group as compared with saline control group. It is concluded that the rhFNHN-29 polypeptide is successfully prepared, this polypeptide can antagonize DIC induced by endotoxin in rats.

[Viral etiology of acute respiratory infection in children from Wenzhou between 2007 and 2008].

OBJECTIVE: To study the viral etiology of acute respiratory infection (ARI)in children from Wenzhou, Zhejiang between 2007 and 2008. METHODS: The nasopharyngeal aspirate samples were obtained from 5 097 hospitalized children with ARI. Seven common respiratory viruses, including respiratory syncytial virus (RSV), influenza virus A and B, parainfluenza viruses 1, 2 and 3 and adenovirus, were detected using direct immunofluorescence. RESULTS: Viral agents were identified in 2 209 cases (43.3%).Of the 2 209, RSV was the most frequent (78.1%), followed by parainfluenza 3 (12.4%), influenza virus A (3.0%), adenovirus (2.8%), parainfluenza 1 (1.7%), influenza B (0.5%) and parainfluenza 2 (0.3%). The infants at ages of <3 months and <6 months had higher detection rate of viruses (53.6% and 49.2%, respectively). A highest detection rate of viruses was found in winter. CONCLUSIONS: RSV is the leading pathogen of ARI in children from Wenzhou, Zhejiang between 2007 and 2008. The children at age of less than 6 months are susceptible to respiratory viruses. The viral activity peaks in winter.

[Association of tumor necrosis factor-alpha-308G/A and interleukin-6-174G/C gene polymorphisms with the susceptibility of respiratory syncytial virus bronchiolitis].

OBJECTIVE: To study the association of tumor necrosis factor (TNF)-alpha-308G/A and interleukin (IL)-6-174G/C gene polymorphisms with the susceptibility of respiratory syncytial virus (RSV) bronchiolitis. METHODS: Polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP) was used to identify the polymorphisms at position 308 of the TNF-alpha promoter and -174 of the IL-6 promoter in 150 children with RSV bronchiolitis and 120 healthy children (control group). RESULTS: The frequency of GG, GA and AA genotypes of TNF-alpha-308 was 68.0%, 28.0% and 4.0% respectively in the RSV group, while that was 80.8%, 19.2% and 0 respectively in the control group. There were significant differences in the genotype frequencies between the two groups (chi2=5.665, p<0.05). The frequency of G and A alleles in the RSV group was 82.0% and 18.0% respectively, while that was 90.4% and 9.6% respectively in the control group. The frequency of A allele in the RSV group was significantly higher than that in the control group (chi2=7.726, p<0.05). Compared to the children carrying G allele, the children with A allele demonstrated a 2.071-fold increased risk of RSV bronchiolitis (OR=2.071, p<0.05). Only GG genotype was detected at position -174 of IL-6 in both groups. CONCLUSIONS: TNF-alpha-308A is associated with the susceptibility of RSV bronchiolitis and it might be an important candidate gene for the development of RSV bronchiolits. IL-6-174G/C gene polymorphism is not found in children in Wenzhou area.

[Recombinant polypeptide of N-terminal heparin-binding domain of fibronectin antagonizes hepatic failure induced by endotoxin in mice].

OBJECTIVE: To study the preventive effect of recombinant polypeptide of N-terminal heparin-binding domain of fibronectin on hepatic failure induced by endotoxin in mice. METHODS: The 40 hepatic failure Balb/C mice were established by intraperitoneal injection of lipopolysaccharide (LPS) and d-galactosamine (GalN). The mice were randomly divided into two groups, one for polypeptide treatment, the othe for saline treatment.Another 20 mice were used as normal control. Half hour prior to, 1, 2, and 3 hours after injection of LPS and GalN, the rhFNHN-29 polypeptide (10 mg/kg) was injected through the tail vein of mice. The same volume of saline was given to the saline treated group and the normal control group.Six hours after the injection of LPS and GalN, 250 microl blood was taken from the eye vein of each mouse for plasma TNFalpha testing, and 72 hours after the injection, mortality rates of the mice of different groups were observated. The liver, lung, heart, kidney, and brain tissues of the survival mice were examined for histopathology after 72 hours. The Liver tissue was also examined for electron micrograph and for mRNA expression of TNFalpha, IL-1beta, IL-6 by RT-PCR. RESULTS: The 72 hours mortality rates in saline-treated and polypeptide treated-mice were 70% and 15% respectively (P < 0.01). The histopathology showed that necrosis occurred less on the hepatocytes of polypeptide treated mice than on the saline treated ones. The ultrastructure of hepatocyte under the electron microscope showed that cell apparatus of saline treated mice were destroyed and cytoplasm become loose. The expression level of TNFalpha, IL-1beta, IL-6 mRNA on hepatocytes in polypeptide treated mice was significantly lower (1.26 +/- 0.37, 0.98 +/- 0.21, 0.43 +/- 0.17, 87.43 +/- 16.7 respectively) than that in the saline treated ones (1.98 +/- 0.56, 1.24 +/- 0.35, 0.64 +/- 0.25 and 236.11 +/- 32.7, respectively) (P < 0.01). Similarly, the plasm TNFalpha level (87.43 +/- 16.7) in polypeptide treated group was significantly lower than that (236.11 +/- 32.7) in the saline treated group (P < 0.01). CONCLUSION: The rhFNHN-29 polypeptide can prevent and treat hepatic failure induced by endotoxin. The mechanism by which the polypeptide takes the effect may involve its ability to down-regulate expression of those inflammation factors such as TNFalpha, IL-1beta, IL-6.