Objective:To investigate the distribution and expression of the cyclooxygenase-2(COX-2)in lung tissues and the effect of its inhibition in bleomycin(BLM)-induced pulmonary fibrosis in rats.Methods:72 male SD rats were randomly divided into three groups:the control group,model group and celecoxib group.Immunohistochemical method was used to detect the distribution of COX-2 in the stage of acute pulmonary alveolitis.HE and Masson staining methods were used to evaculate the degree of alveolitis and pulmonary fibrosis.Results:(1)In early stage of pulmonary fibrosis except in the control group,COX-2 expression was found in small bronchial epithelial cells in the other two groups,especially the model group.(2)Compared with the control group,the other two groups had alveolitis(P

Objective A morphometric study was carried out on GABAergic neurons and astrocytes in primary auditory cortex of 4 young cats (1-3 years old, 20-2.5 kg) and 4 old cats (10-13-year-old, 3-3.5 kg), in order to explore the age-related changes and the probably physical effects underlying these changes. Methods The same regions of the primary auditory cortex (AI) were comparatively studied. Nissl staining and immunohistochemical methods were applied to explore the differences of GABAergic neurons and astrocytes(specifically labeled by GFAP as a mark as antibody) in the primary auditory cortex(Al) between young and old cats. Serial cross-sections from two age groups were stained with Nissl for layer structure observation and demonstration of neurons; Adjacent sections were stained with anti_GABA immunohistochemistry for localizing and characterizing GABAergic and with anti-GFAP for astrocytes in AI. Results The diameter, density and number of GABAergic neurons in Al of old cats declined significantly compared with the counterpart in young cats, especially in layers Ⅱ,Ⅲ of AI; but the change in GFAP-IR cells was reversed: old cats showed clearly larger number and stronger staining of GFAP-IR cells than that in young cats. The GFAP-IR cell bodies and dendrites swelled obviously in old cats, especially in layer Ⅰ of AI and in white matter. Conclusion The result presented a direct morphological evidence for weakness of GABAergic intracortical inhibition, which is considered to be a possible mechanism for age-related acoustic function decline. The enhanced function of astrocytes may play an important role in decreasing the GABA content in AI of the old individuals.

Objective To investigate the clinical value of multiband mucosectomy (MBM) for the treatment of early esophageal cancer and precancerous lesions.Methods The clinical data of 43 patients with early esophageal cancer or precancerous lesions (including 13 cases of early esophageal cancer and 30 cases of precancerous lesion) who were admitted to the Tengzhou Central People's Hospital from January 2010 to January 2011 were retrospectively analyzed.A total of 51 lesions were confirmed by preoperative endoscopy and mucosal biopsy,and then were treated by MBM.Results A total of 52 MBM procedures were carried out,and 180 lesions were resected.The sizes of the lesions ranged from 10 mm × 8 mm to 25 mm × 23 mm,and the mean operation time was (37 ± 5)minutes.The complete resection rate was 92.2％ (47/51).The submucosal layer of 1 patient was invaded by squamous cell carcinoma postoperatively,and the patient was treated by operation;1 patient wth squamous epithelium carcinoma was treated by radiotherapy at 2 weeks after MBM ; the other 2 lesions were treated by argon plasma coagulation (APC).Four lesions with bleeding were detected during 52 MBM procedures,and no esophageal perforation was detected during MBM.Early complications consisted of delayed bleeding (1 patient,cured by APC) and slight esophageal stenosis (1 patient,recovered at postoperative month 6).The accurate rate of pathological examinations was 51.0％ (26/51).Forty-three patients were followed up for 1 year,3 patients with tumor recurrence were treated by MBM or APC.No death occurred during the followup.Conclusion MBM is a safe and effective technique for the treatment of early esophageal cancer and precancerous lesions.

BACKGROUND: The study of tissue-engineered cartilage with predetermined shaping and regeneration has provided novel ideas and techniques for repair of laryngeal cartilage erosion; however, due to the special natures of the morphology, location and function of laryngeal cartilage, tissue engineering research has not, to date, exhibited its ful advantages in the reconstruction of laryngeal cartilage. OBJECTIVE:To explore the feasibility of building tissue-engineered larynx-shaped cartilage using poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHH) as a scaffold filed and encapsulated with pedicled myofascial flaps. METHODS:Porous PHBHH was prepared and formed into a holow like larynx-shape, and the cel PHBHH composites were cultured for 1 weekin vitro prior to implantationin vivo. The cel-PHBHH composite was filed and encapsulated with myofascial flaps with the pedicle forin situ implantation in nine rabbits as experimental group. PHBHH scaffold with no chondrocytes was implanted alone in three rabbits as control group. Cartilage regeneration was assessed at 6, 12 and 18 weeks after surgery through morphological observation, histological and immunohistochemical detection. RESULTS AND CONCLUSION: In the experimental group, the shape and porosity (> 90%) of the material were ideal, the cels exhibited good adhesion with the material and the blood supply within the myofascial flap with pedicle was rich for effective filing and encapsulation of the cel PHBHH composite. Tissue-engineered laryngeal cartilage with the holow, semi-trumpet shape was idealy formed at 6 weeks after the surgery. Further maturation of the cartilage was observed at 12 and 18 weeks after the surgery. However, there was no cartilage tissue in the control group. This study shows that PHBHH is a suitable material for the formation of a holow, semi-trumpet shape with good celular compatibility. Myofascial flap filing and encapsulating can be used to build tissue-engineered laryngeal cartilage with a holow, semi-trumpet shape.

Objective To explore the effect of DR on the expression of inhibitory neurotransmitter ,γ-aminobutyric acid ( GABA) and brain derived neurotrophic factor ( BDNF) in the primary visual cortex of young cats .Methods Totally 6 cats were wsed in this study . Nissl staining was used for cortical layer identification and cell counting . Immunohistochemical techniques were utilized to label GABA-and BDNF-positive neurons .Sections were observed under an Olympus light microscope and photographed with a digital camera .The cell density and absorbance of immunoreactivity were measured with Image-Pro Express 6.0 softwares .Results Our results showed that the mean density of Nissl-stained neurons in the primary visual cortex ( V1) of the DR group showed no significant difference from that of the control group . However , the mean density of GABA-immunoreactive neurons in each cortical layer of V 1 in DR cats was significantly higher than that in control cats .The mean immunoreactive intensity of GABA-positive neurons , as indicated by the average absorbance , increased significantly in DR cats relative to control ones . Accompanied with the elevation of GABA expression, DR enhanced BDNF expression in V1, as indicated by an increased mean density of BDNF-positive neurons and BDNF-immunoreactive average absorbance in DR cats relative to controls .Conclusion These results indicate that dietary restriction leads to a concurrent expression upregulation of GABA and BDNF , which may likely compensate for brain functional degradation during senescence and thus delay aging process .

Objective To determine the serum levels of tumor necrosis factor (TNF)-α,interleukin (IL)-17,IL-22 and IL-17F in patients with palmoplantar pustulosis (PP),and to estimate their relationship with disease activity in PP.Methods Venous blood samples were collected from 30 patients with PP at both active stage and stationary stage and from 20 healthy human controls.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine the serum levels of TNF-α,IL-17,IL-22 and IL-17F.The paired Wilcoxon signed rank test was carried out to compare the serum levels of cytokines between patients at active stage and at stationary stage,and the Mann-Whitney U test to compare those among different groups.Results The median serum levels of TNF-α,IL-17 and IL-22 in patients with PP at active stage were 186.35 (range,113.48-412.69) ng/L,420.45 (range,278.55-748.73) ng/L and 106.48 (range,69.13-251.86) ng/L respectively,significantly higher than those at stationary stage (42.52(18.83-95.37) ng/L,48.11 (36.43-80.04) ng/L,20.32 (10.55-48.75) ng/L,respectively,all P ＜ 0.05) and those in the controls (24.30 (12.0-61.56) ng/L,10.49 (6.24-24.44) ng/L,2.58 (1.41-5.78) ng/L,respectively,all P ＜ 0.05).Moreover,the patients at stationary stage showed a significant elevation in serum levels of TNF-α,IL-17 and IL-22 compared with the controls (u =2.71,3.53,2.18,respectively,all P ＜ 0.05).No statistical difference was noted in the serum level of IL-17F among the patients at different stages and controls (P ＞ 0.05).Conclusion The circulating levels of TNF-α,IL-17 and IL-22 were associated with disease activity in PP,hinting that they may be involved in the development of PP.

ObjectiveTo explore the difference in quality of life in patients with gastrointestinal cancer between those who knew cancer diagnosis and those who did not.Methods 130 patients with recently diagnosed gastrointestinal cancer in 4 general hospitals in Tangshan were interviewed. Patients were categorized into two groups:one group was those who knew their diagnosis(n=62) and the other group was those who did not ( n =68).The Chinese version of European Organization for Research and Treatment (EORTC) quality of life (QOL)questionnaire (QLQ-C30) was used.ResultsThe comparison of quality of life between patients who knew their diagnosis and those who did not indicated that the score of global quality of life and other functional scales including physical,emotional,role and cognitive functioning did not show significant difference(P ＞ 0.05 ).Compared with those who did not know cancer diagnosis,patients who were aware of diagnosis showed a significant lower degree of social functioning ( (45.8 ± 24.2 ) vs ( 73.0 ± 16.2 ) ),a significant higher degree of fatigue ( ( 55.8 ±26.2) vs (41.8 ±10.2) ) and financial difficulties ((38.2 ±35.1) vs (12.00 ±10.0) ) (P＜0.01).In multiple regression analysis,physical functioning and symptom of fatigue had statistically significant differences (P＜ 0.05 ).ConclusionThe knowledge of cancer diagnosis does not contribute to the quality of life in patients with gastrointestinal cancer.

Objective:To assess the prognostic value of modified Charlson comorbidity index (mCCI) combined with serum albumin for long-term prognosis in peritoneal dialysis (PD) patients.Methods:From January 1, 2007 to June 30, 2015, patients who started PD in Nanjing Drum Tower Hospital were enrolled in this retrospective cohort study. Clinical data including gender, age, underlying diseases, laboratory examination and prognosis were collected. The mCCI at the beginning of PD was calculated. Whether the duration of PD exceeded 5 years was used as an indicator to evaluate the prognosis. The patients were divided into≥5 years group and<5 years group according to the duration of PD, and the data were compared between the two groups. Cox regression model was constructed to analyze the influencing factors of all-cause death in PD patients. Multivariate logistic regression model and receiver operating characteristic (ROC) curve were used to analyze the predictive value of mCCI and serum albumin levels on whether patients could maintain long-term PD.Results:Of the 183 patients included [males 106(57.9%), females 77(42.1%); (53.35±16.50) years old; 162 cases (88.5%) with hypertension, 55 cases (30.1%) with diabetes], 97 cases had PD duration for ≥5 years and 86 cases less than 5 years. The overall 5-year technical survival rate was 65.1%. At the beginning of PD, compared with the dialysis age≥5 years group, the patients in the dialysis age less than 5 years group had older age, higher mCCI, lower serum albumin level, and higher C-reactive protein (CRP) level (all P<0.05), but there were no significant differences in gender, education level, electrolyte, mean arterial pressure, high densitv lipoprotein (HDL), low-density lipoprotein (LDL) and PD adequacy index between the two groups (all P>0.05). Multivariate logistic regression analysis showed that increased age ( OR=1.022, 95% CI 1.000-1.043, P=0.046), increased mCCI ( OR=1.620, 95% CI 1.300-2.018, P<0.001) and decreased serum albumin ( OR=0.807, 95% CI 0.730-0.893, P<0.001) were independent predictors for the duration of PD<5 years. ROC curve analysis showed that the area under ROC curves ( AUC) of mCCI, serum albumin level and combined prediction probability of the two for the duration of PD<5 years were 0.647(95% CI 0.568-0.727), 0.655(95% CI 0.577-0.734), and 0.767(95% CI 0.700-0.835), respectively, indicating that the accuracy of combined parameters to predict survival outcome was higher than that of any single parameter. Multivariate Cox analysis showed that increased age ( HR=1.073, 95% CI 1.046-1.100, P<0.001), increased mCCI ( HR=1.198, 95% CI 1.044-1.375, P=0.010) and decreased serum albumin ( HR=0.904, 95% CI 0.843-0.969, P=0.004) were independent influencing factors for all-cause death in PD patients. Conclusions:Old age, high mCCI and low serum albumin level are influencing factors for dialysis age<5 years and all-cause death in PD patients. mCCI combined with serum albumin level can improve the accuracy of predicting the long-term dialysis in PD patients.

Coenzyme Q10 (CoQ10) is a lipophilic antioxidant that improves human immunity, delays senility and enhances the vitality of the human body and has wide applications in pharmaceutical and cosmetic industries. Microbial fermentation is a sustainable way to produce CoQ10, and attracts increased interest. In this work, the native CoQ8 synthetic pathway of Escherichia coli was replaced by the CoQ10 synthetic pathway through integrating decaprenyl diphosphate synthase gene (dps) from Rhodobacter sphaeroides into chromosome of E. coli ATCC 8739, followed by deletion of the native octaprenyl diphosphate synthase gene (ispB). The resulting strain GD-14 produced 0.68 mg/L CoQ10 with a yield of 0.54 mg/g DCW. Modulation of dxs and idi genes of the MEP pathway and ubiCA genes in combination led to 2.46-fold increase of CoQ10 production (from 0.54 to 1.87 mg/g DCW). Recruiting glucose facilitator protein of Zymomonas mobilis to replace the native phosphoenolpyruvate: carbohydrate phosphotransferase systems (PTS) further led to a 16% increase of CoQ10 yield. Finally, fed-batch fermentation of the best strain GD-51 was performed, which produced 433 mg/L CoQ10 with a yield of 11.7 mg/g DCW. To the best of our knowledge, this was the highest CoQ10 titer and yield obtained for engineered E. coli.
Alkyl and Aryl Transferases ; genetics ; Bacterial Proteins ; genetics ; Batch Cell Culture Techniques ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Gene Deletion ; Industrial Microbiology ; Metabolic Engineering ; Rhodobacter sphaeroides ; enzymology ; genetics ; Ubiquinone ; analogs & derivatives ; biosynthesis ; Zymomonas ; genetics

β-carotene belongs to carotenoids family, widely applied in pharmaceuticals, neutraceuticals, cosmetics and food industries. In this study, three key genes (dxs, idi, and crt operon) within β-carotene synthetic pathway in recombinant Escherichia coli strain CAR005 were modulated with RBS Library to improve β-carotene production. There were 7%, 11% and 17% increase of β-carotene yield respectively after modulating dxs, idi and crt operon genes with RBS Library, demonstrating that modulating gene expression with regulatory parts libraries would have more opportunities to obtain optimal production of target compound. Combined modulation of crt operon, dxs and idi genes led to 35% increase of β-carotene yield compared to parent strain CAR005. The optimal gene expression strength identified in single gene modulation would not be the optimal strength when used in combined modulation. Our study provides a new strategy for improving production of target compound through modulation of gene expression.
Escherichia coli ; metabolism ; Gene Expression ; Gene Library ; Operon ; beta Carotene ; biosynthesis