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Objective: To investigate the role of an inflammatory microenvironment induced by Porphyromonasgingivalis ( P. gingivalis ) in the occurrence of esophageal squamous cell carcinoma (ESCC) in mice. Methods: A total of 180 C57BL/6 mice were randomly divided into 6 groups, i.e. control group, P. gingivalis group, 4NQO group, 4NQO + P. gingivalis group, 4NQO + P. gingivalis + celecoxib group, and 4NQO + P. gingivalis + antibiotic cocktail (ABC, including metronidazole, neomycin, ampicillin, and vancomycin) group, with 30 mice in each group, using the random number table. All mice were normalized by treatment with ABC in drinking water for 2 weeks. In the following 2 weeks, the mice in the control group and the P. gingivalis group were given drinking water, while the other 4 groups were treated with 30 μg/ml 4NQO in the drinking water. In weeks 11-12, the mice in the P. gingivalis group, the 4NQO + P. gingivalis group, the 4NQO + P. gingivalis + celecoxib group, and the 4NQO + P. gingivalis + ABC group were subjected to ligation of the second molar in oral cavity followed by oral P. gingivalis infection thrice weekly for 24 weeks in weeks 11-34. In weeks 13-34, the mice in 4NQO + P. gingivalis +celecoxib group and 4NQO + P. gingivalis + ABC group were administered with celecoxib and ABC for 22 weeks, respectively. At the end of 34 weeks, gross and microscopic alterations were examined followed by RT-qPCR and immunohistochemistry to examine the expression profiles of inflammatory- and tumor-molecules in esophagi of mice. Results: At 34 weeks, 4NQO treatment alone did not affect the foci of papillary hyperproliferation, diseased area, and the thickness of the esophageal wall, but significantly enhanced the foci of hyperproliferation (median 1.00, P ＜0.05) and mild/moderate dysplasia (median 2.00, P ＜0.01). In addition, the expression levels of IL-6 [8.35(3.45,8.99)], IL-1β [6.90(2.01,9.72)], TNF-α [12.04(3.31,14.08)], c-myc [2.21(1.80,3.04)], pSTAT3, Ki-67, and pH2AX were higher than those in the control group. The pathological changes of the esophageal mucosa were significantly more overt in the 4NQO + P. gingivalis group in terms of the foci of papillary hyperproliferation (median 2.00), diseased area (median 2.51 mm 2 ), the thickness of the esophageal wall (median 172.52 μm), the foci of hyperproliferation (median 1.00, P ＜0.05), and mild/moderate dysplasia (median 1.00, P ＜0.01). In mice of the 4NQO + P. gingivalis group, the expression levels of IL-6 [12.27(5.35,22.08)], IL-1β [13.89(10.04,15.96)], TNF-α [19.56(6.07,20.36)], IFN-γ [11.37(8.23,20.07)], c-myc [2.62(1.51,4.25)], cyclin D1 [4.52(2.68,7.83)], nuclear pSTAT3, COX-2, Ki-67, and pH2AX were significantly increased compared with the mice in the control group. In mice of the 4NQO + P. gingivalis group, the diseased area, invasive malignant foci as well as pSTAT3 and pH2AX expression were significantly blunted by celecoxib. Treatment with ABC markedly reduced the papillary hyperproliferative foci, invasive malignant foci, and pSTAT3 expression but not pH2AX. Conclusions: P. gingivalis promotes the occurrence of esophageal squamous cell carcinoma in mice by inducing an inflammatory microenvironment primed with 4NQO induced DNA damage. Clearance of P. gingivalis with ABC or anti-inflammatory intervention holds promise for prevention of esophageal squamous cell malignant pathogenesis via blockage of IL-6/STAT3 signaling and amelioration of inflammation.

This study aimed to investigate the effect and possible mechanism of Bidens pilosa decoction on non-alcoholic fatty liver disease(NAFLD) induced by high fat and high glucose in mice. Bald/c mice were randomly divided into normal group, model group, metformin(200 mg·kg~(-1)) treatment group, Bidens pilosa decoction(10 g·kg~(-1)) treatment group, metformin and B. pilosa decoction(100 mg·kg~(-1)+5 g·kg~(-1)) treatment group. Except for the normal group, mice in the other four groups were fed with high-fat and high-glucose diet for 8 weeks to establish the non-alcoholic fatty liver model. After 4 weeks of treatment, blood was collected from the eyeballs, the mice were sacrificed, and relevant indicators were detected. The results showed that compared with the model group, blood lipid and blood glucose levels of each treatment group were significantly lower(P<0.05); HE staining results showed that liver pathological damage in each treatment group was significantly improved; oil red O staining results showed fat distribution in each treatment group significantly reduced(P<0.01); immunohistochemical staining showed that glucose regulated the protein expression of protein 78(GRP78) in liver tissues of each treatment group was also significantly reduced(P<0.01); Western blot results showed that endoplasmic reticulum stress signal pathway-related factors GRP78, phosphorylated-protein kinase R-like ER kinase(p-PERK), eukaryotic translation-initiation factor 2α(eIF2α), activating transcription factor 4(ATF4), C/EBP homologous protein(Chop), inositol requiring 1α(IRE1α), and cleaved-cysteinyl aspartate specific proteinase 12(cleaved-caspase-12) were significantly reduced(P<0.01). The results of the combined drug treatment group were better than those of the single drug treatment group. These results showed that B. pilosa decoction had the effect in improving non-alcoholic fatty liver, and its mechanism may be related to the down-regulation of the expression of endoplasmic reticulum stress(ERS)-related factors, and the reduction of the apoptosis of hepatocytes caused by ERS and the down-regulation of blood lipid and blood glucose levels.

Objective: To explore the relationship between physiological parameters changes and severe heatstroke induced by 5-km armed cross-country training., Methods: A total of 521 male officers and soldiers from a special team who participated in the summer training of 5-km armed cross-country training from year 2016 to 2017 were enrolled. All trainees participated in 5-km armed cross-country training in high temperature and humidity environment of ambient temperature > 32 centigradeand (or) relative humidity > 65%. The trainees were divided into two groups according to the incidence of severe heatstroke in the course of training. The age, enlistment time, constitution score, body mass index (BMI), external environment (ambient temperature, relative humidity, wind speed, heat index) of trainees of the two groups, and the change rates of arterial blood oxygen saturation (SaO 2 ), body temperature, pulse and blood pressure within 5 minutes after the 5-km armed cross-country training were compared between the two groups. The risk factors of severe heatstroke were screened by two classified Logistic regression analysis, and the predictive value of various risk factors of severe heatstroke was analyzed by the receiver operator characteristic curve (ROC)., Results: In 521 trainees of 5-km armed cross-country training, 29 trainees suffered from severe heatstroke accounting for 5.57%. There was no significant difference in the age, enlistment time, constitution score, BMI, or external environment during 5-km armed cross-country training between severe heatstroke group and non-severe heatstroke group. Compared with those without severe heatstroke, the descending rates of body temperature, pulse, blood pressure and SaO 2 increased rate within 5 minutes after 5-km armed cross-country training of severe heatstroke trainees were significantly decreased [temperature descending rate: (0.67±0.30)% vs. (1.43±1.28)%, pulse descending rate: (7.53±5.21)% vs. (13.48±8.07)%, blood pressure descending rate: (9.28±6.84)% vs. (19.42±7.73)%, SaO 2 increased rate: (0.51±0.39)% vs. (1.50±1.43)%, all P < 0.01]. Two classification Logistic regression analysis showed that the temperature descending rate [odds ratio (OR) = 0.485, 95% confidence interval (95%CI) = 0.289-0.817], pulse descending rate (OR = 0.903, 95%CI = 0.845-0.965), blood pressure descending rate (OR = 0.841, 95%CI = 0.790-0.896), and SaO2 increased rate (OR = 0.421, 95%CI = 0.250-0.711) were the risk factors for severe heatstroke during 5-km armed cross-country training (all P < 0.01). ROC curve analysis showed that temperature descending rate [area under ROC curve (AUC) = 0.659, 95%CI = 0.604-0.714], pulse descending rate (AUC = 0.730, 95%CI = 0.762-0.900), blood pressure descending rate (AUC = 0.831, 95%CI = 0.659-0.801), SaO 2 increased rate (AUC = 0.711, 95%CI = 0.655-0.767) could be used for the incidence of severe heatstroke prediction during 5-km armed cross-country training (all P < 0.01), and the predicted value was the same., Conclusions: Under the same conditions, the severe heatstroke during 5-km cross-country training is closely related to the descending rates of body temperature, pulse, and blood pressure as well as SaO 2 increased rate within 5 minutes after the training, whose predictive values for severe heatstroke were the same.

Objective: To explore the clinical characteristics and treatments of patients with idiopathic sudden sensorineural hearing loss（ISSHL） with or without vertigo. Method: One hundred and twenty ISSHL cases were divided into vertigo group ( n =36) , without vertigo group ( n =84) , and with benign paroxysmal positional vertigo group ( n =15). All patients were in regular treatment. Besides, according to the types of BPPV, patients do the Epley maneuver or Barbecue roll maneuver. We summarized the result and treatment of the patients. Result: The audiometric curve of ISSHL with vertigo were mainly at flat type. After treatment of the ISSHL patients were better than the patients with vertigo in the degrees of hearing loss . Furthermore, the rate of the patients of marked efficiency, efficiency and total efficiency of ISSHL was lower than the ones without.The patients with BPPV, including 12 cases of posterior semicircular canal and the 3 cases of lateral semicircular canal, were all ipsilateral. Conclusion: ISSHL with vertigo group lost hearing is severer than ISSHL without vertigo. Thus the hearing and the treatment effect were worse.The symptoms without vertigo in ISSHL were better than the patients with vertigo., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: To investigate the expression of serum TM and MPO in adult patients with obstructive sleep apnea hypopnea syndrome(OSAHS). Method: Ninety OSAHS patients who confirmed by PSG as OSAHS group. According to AHI, the patients were divided into 3 groups include heavy, medium and light, and 30 healthy outpatients were control group. The serum TM and MPO were determined by ELISA; TM and MPO were measured after comprehensive treatment in patients with severe OSAHS, and the correlation between TM, MPO and PSG were analyzed. Result: ①With the severity of OSAHS patients increased, the serum levels of TM and MPO increased gradually（F=20.761,21.433； P <0.01). There was no significant difference about the concentration of TM and MPO between light and control group（ P >0.05）；The concentration of TM and MPO was significantly higher in heavy and medium group compared with light and control group（ P <0.01）.②There was no correlation between serum levels of TM, MPO, BMI, age and sex in OSAHS patients( P >0.05). The serum concentration of TM was positively related to MPO. The serum concentrations of TM and MPO were positively correlated with AHI, but negatively with LSaO₂ ( P <0.05). ③LSaO₂ was significantly increased, AHI and peripheral blood TM, MPO levels were significantly reduced in thirty severe OSAHS cases received the combined treatment( P <0.01). Conclusion: The increase of TM and MPO concentration in peripheral blood is one of characteristics of cardiovascular damage in patients with OSAHS. Combined detection of serum TM and MPO concentrations in patients with the disease is helpful for evaluation and risk assessment of cardiovascular disease., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Aim: To clone human high mobility guoup box1 A box (HMGB1 A box) and express it in escherichia coli effectly, investigate the inhibit effection of the purpose protern to the activation of monocytes stimulated by immunocomplex., Methods: According to human HMGB1 gene order which was optimized by our laboratory the PCR primer was designed which containing restriction enzyme cutting site. The HMGB1 A box gene was cloned following the whole gene synthesis template of human HMGB1, then the PCR product was inserted into clone vector pMD19-T. The positive colone was identified by colony PCR, zymography analysis and DNA sequencing. Recombinant colne vector was digested by restriction enzymes Nde I and Xho I and separated by agarose gel electrophoresis, then the fragment was inserted into the corresponding sites of expression vector pQE-T7-2. The positive recombinant expression vector was identified by colony PCR and the recombinant strains was induced by IPTG, then the purpose protein was identified by SDS-PAGE and Western blot. The recombinant protein of human HMGB1 A box was purificated by Ni(2+)-NTA chromatography and the inhibit effection of the purpose protern to the activation of monocyte stimulated by immunocomplex was identified by RT-PCR., Results: We acquired expression strains of recombinant human HMGB1 A box, the target protein account for up to 40% of the whole protein of E.coli. Western blot showed recombinant protein can specificly reacted with anti-human HMGB1 polyclonal antibody and anti-His-Tag polyclonal antibody.The purpose protein was found more than 90% after purified, and can effectively inhibit the production of BAFF, IFN-gamma and TNF-alpha in monocyte which were induced by IC., Conclusion: A recombinant bacterial strain for expressing human HMGB1A box with biological activities was constructed successfully.

Objective: To investigate the association of killer cell immunoglobulin-like receptors (KIR) genotype and haplotype with ankylosing spondylitis (AS)., Methods: Peripheral blood samples were collected from 105 AS patients, 62 patients of osteoarthritis (OA), and 412 randomly selected healthy controls. Polymerase chain reaction with sequence-specific primers (PCR-SSP) was used to detect the KIR genotype and haplotype., Results: The genotype frequency of 3DL3-2DL3-2DL1-2DP1-2DL4-3DL1-2DS4-3DL2 (6.67%) was significantly lower in the AS patients than in the control subjects (20.15%) and OA patients (17.74%, P = 0.001, 0.037 respectively). The genotype frequency of 3DL3-2DL3-2DL2-2DL1-2DP1-2DLA-3DL1-2DL5-2DS1-2DS2-2DS3-2DS4-2DS5-3DS1-3DL2 and 3DL3-2DL3-2DL2- 2DL1-2DP1-2DL4-3DL1-2DL5-2DS1-2DS4-3DL2 of the AS patients (9.52%, 5.71%)was significantly higher than that of the controls(2.18%, 0.49%; P = 0.001, 0.001), and these two genotypes were not detected in the OA patients. There were not significant differences in the haplotypes A and B among the AS patients, OA patients, and healthy controls., Conclusion: KIR genotypes may be associated with the susceptibility to AS.

Taking two peanut varieties with different drought-resistance Luhua 11 and Nongda 818 as test crops, the effects of different irrigation amount at seedling stage on their physiological characteristics and yield were studied from 2003 to 2004. The results showed that with decreasing irrigation amount, the leaf photosynthesis rate of test varieties decreased, while malondialdeyde (MDA) content increased. Watering 60-80 mm ( suitable drought) enhanced the activities of superoxide dismutase (SOD) , peroxidase (POD) and catlase (CAT) , and increased the content of soluble protein. After re-watering by the end of treatments, the activities of SOD, POD and CAT and the contents of soluble protein and MDA decreased significantly, while photosynthesis rate increased obviously. The pod and kernel yield decreased with decreasing irrigation amount, and Luhua 11 had a greater loss than Nongda 818, indicating that Nongda 818 was more drought-tolerant than Luhua 11. It was suggested that under water-saving culture, the irrigation amount at seedling stage could not be less than 80 mm for Luhua 11, and less than 60 mm for Nongda 818.

To clone human interleukin-26 (hIL-26) and express it in E. coli efficiently. Two pairs of primers were synthesized according to the hIL-26 gene reported on GenBank. The hIL-26 gene was cloned by nest PCR following the first round RT-PCR from human peripherial blood monocytes total RNA, and then the PCR product was cloned into pMD18-T vector. Colony PCR, restriction analysis and sequence analysis showed that the gene cloned was the same as the reported hIL-26. The recombinant was cut with BamHI and EcoR I to obtain the hIL-26 fragment, and then the fragment was inserted into pBV220 which was cut with the same enzymes. The recombinant expression vector was induced to express hIL-26 at 42 degrees C, SDS-PAGE analysis showed that the recombinant protein accounted for up to 20% of the whole protein of E. coli, and the protein was also confirmed by Western blotting. Purity of the protein was found to be above 90% after purified with molecular sieve. After renaturalized with glutathione buffer, the promoting effect of it on the production of IFN-y in PBMC was detected by RT-PCR. A recombinant bacterial strain for expressing hIL-26 with biological activity was constructed successfully.

Aim: To construct an eukaryotic co-expression plasmid pSLC-IRES-IL-2, and to express it in COS-7 cells., Methods: Human IL-2 and SLC genes were cloned by RT-PCR and PCR, respectively, and then the eukaryotic expression plasmid pSLC-IRES-IL-2 was constructed.The constructed plasmid was transfected into COS-7 cells by electroporation method. The expression of SLC and IL-2 was detected by Western blot., Results: SLC and IL-2 in the culture supernatant and lysate of transfected COS-7 cells were detected by Western blot. The relative molecular masses of the expressed products were consistent with the theoretical values., Conclusion: The SLC and IL-2 co-expression plasmid is successfully constructed and expressed in COS-7 cells, which offers a pathway for research on gene therapy of tumors.

Objective: To study the curative effect of grafting great auricular nerve with pediculated fascial tube in defective damage of facial nerve., Methods: All the 7 patients in this study were treated by grafting great auricular nerve covered by pediculated fascial tube near facial nerve trunk., Results: Four cases with otogenic facial paralysis had grade III - IV recovery of facial nerve function from two to two and half years after the nerve grafting operation. Three patients post-traumatic facial paralysis had grade III recovery of facial nerve function 2 years after the nerve grafting operation., Conclusions: The grafting of pediculated fascial tube surrounded great auricular nerve can provide a biological environment with better blood supply for the plerosis and regeneration of nerve and can accelerate the functional recovery of nerves after the nerve grafting.