AIM: To investigate the effect of erythromycin on the level of transforming growth factor-?1(TGF-?1) and ?-glutaglutamylcysteine synthetase(?-GCS) in smoking rats,and to explore the antioxidate therapeutic role of erythromycin in chronic obstructive pulmonary disease.METHODS: Wistar rats were exposed to cigarettes smoking to establish the model.After passive smoking for 4 weeks,erythromycin intragastric intervention was administered continuously for 8 weeks.The expiratory airway resistance and lung compliance were assessed and the expression levels of TGF-?1 and ?-GCS proteins(and the mRNA) in airway endothelial cells and alveolar macrophages were observed respectively by immunohistochemical,immunocytochemical and(in situ) hybridization.RESULTS: The expiratory airway resistance was increased and the lung compliance was degraded significantly in smoking group and erythromycin group,compared to control group.In erythromycin group,the airway resistance was lower and the lung compliance was higher than that in smoking group(P

Objective To explore the effect of purification on monoclonal antibody (MAb) against PGRP by Protein A-Sepharose affinity chromatography, and to provide some based data for the purification of other antibody using the same method. Methods The ascites which include MAb was purified by Protein A-Sepharose affinity chromatography. The purity and activity of MAb was tested by SDS-PAGE and ELISA. The biological function was identified by flow cytometer and immunohistochemistry. Results The average concentration of protein in ascites before purification is 23.62 mg/ml. Before and after purification, the total protein is 148.79 mg and 146.67 mg, respectively. The recovery coefficient of protein is 98.58%. The concentration of MAb in ascites is 5.21 mg/ml averagely. The MAb purity is more than 95 %. The immunoactivity of purified antibody is higher than that of unpurified antibody. Conclusion The purity of MAb against PGRP purified by Protein A-Sepharose affinity chromatography is very high. The immunoactivity of purified antibody is higher than that of unpurified antibody. So the ProteinA-Sepharose affinity chromatography is a rapid, convenient and reliable method for the purification of MAb Against PGRP.

Objective To explore the optimal method of treatment for radiation proctitis by keeping coloclysis with traditional Chinese medicine (TCM), including administation temperature, infusion time, dosage and catheter depth. Methods The orthogonal experimental design was adopted. Sixty-three patients with radiation proctitis were randomly divided into 9 groups, and they were under enema for 6 weeks according to different test conditions. TCM syndrome score, radiation injury effect and Karnofsky scores were set as evaluation indexes. An orthogonal design and analysis of variance were conducted for optimization. The best technical schemes for traditional Chinese herb in treating radiation proctitis were obtained. Results The obtained optimum methods are:drug temperature of (39±0.5)℃, infusion time of 30 minutes, dosage of 100 mL, catheter depth of 20 cm. Conclusion The optimal scheme of enema for the treatment of radiation proctitis is reasonable and feasible.

Objective To analyze risk factors for diabetic kidney disease (DKD) in inpatients with type 2 diabetes.Methods A total of 930 inpatients with type 2 diabetes were enrolled in the study and grouped according to different levels of estimated glomerular filtration rate (eGFR),albuminuria,and diabetic retinopathy.Logistic regression analysis was adopted to explore the risk factors for DKD in inpatients with type 2 diabetes.Results (1) The prevalence of albuminuria in patients with type 2 diabetes mellitus was increased with declining eGFR (P ＜ 0.05).(2) The prevalences of DKD and non-diabetic renal disease (NDRD) in patients with type 2 diabetes mellitus were 22.26％ and 8.92％,respectively.Compared with patients with NDRD,patients with DKD had longer diabetic duration,higher levels of systolic blood pressure,serum creatinine,and urinary albumin excretion,and lower levels of hemoglobin[(125.40 ± 21.95 vs 138.18 ± 19.67) g/L],serum albumin[(37.45 ± 5.54 vs 40.55 ± 3.55) g/L],and eGFR[(89.66 (59.10-108.25) vs 103.15 (85.39-114.88) ml · min-1 · (1.73 m2)-1,all P＜0.05].(3) Logistic regression analysis showed that age,diabetic duration,systolic blood pressure,serum uric acid,diabetic retinopathy,and hypertension are the independent risk factors for diabetic kidney disease in inpatients with type 2 diabetes,while serum albumin was the protective factor (all P＜0.01).Conclusions A variety of clinic risk factors were associated with DKD.Better control of blood pressure,serum uric acid,and hypoalbuminemia should be performed to delay the progress of DKD.

Objective The value of pro-gastrin releasing peptide ( PGRP) which is the tumor marker of small cell lung canc-er has become a hot topic in recent years .The research was to build a new enzyme-linked immune sorbent assay ( ELISA) method ai-ming at detecting the concentration of PGRP in patients′serum. Methods We utilized synthetic PGRP epitopes for the screening of the monoclonal antibodies , labeled the screened monoclonal antibodies with horseradish peroxidase by modified sodium iodide method , and then established double antibody sandwich ELISA which could be used to detect the serum concentrations of PGRP in cancer pa -tients. Results We successfully screened E 12 mAb which could be served as the coating antibody and ED 1 mAb as the labeled anti-body.The standard antibody density range of new ELISA was 33 pg/mL~1.7 ×104 pg/mL.The comparison experiments between our method and the commercially available ELISA kit showed no significant difference ( P>0.05).The specificity of our method was 50%, and the sensitivity was 100%, while IBL kit was 92.2% and 100% respectively. Conclusion New ELISA can be used to detect the serum PGRP concentration in patients with small cell lung cancer .

OBJECTIVETo investigate the effects of the DPP4 inhibitor sitagliptin on the expressions of early growth response-1 (Egr-1) and fibronectin in the kidney of ApoE gene knockout mice.

METHODSEight-week-old male ApoE gene knockout mice were randomly divided into sitagliptin + apoE(-/-) group and apoE(-/-) group (n=6), with 6 C57BL mice as the normal control group. After feeding with high-fat diet and drug treatment for 16 weeks, the mice underwent intraperitoneal glucose tolerance test (IPGTT) and were measured for 24-h urinary albumin using ELISA. All the mice were then sacrificed to examine the changes of blood lipid profile and for detection of Egr-1 and fibronectin mRNA and proteins in the renal tissue using real-time PCR and Western blotting.

RESULTSThe mice in both apoE(-/-) group and sitagliptin+apoE(-/-) group all showed prominently increased blood lipids as compared with the control group (P<0.05) without significant differences between the two apoE(-/-) groups. The level of HDL was significantly higher in sitagliptin +apoE(-/-) group than in apoE(-/-) group (P<0.001) and control group (P<0.001). IPGTT showed no significant differences in the levels of blood glucose among the 3 groups. The excretion of urinary albumin was increased in apoE(-/-) group compared with the control group (P<0.01), but was significantly lower in sitagliptin+ apoE(-/-) group than in apoE(-/-) group (P<0.01). Real-time PCR and Western blotting showed significantly decreased mRNA and protein expressions of renal cortical Egr-1 and fibronectin in sitagliptin+apoE(-/-) group compared with apoE(-/-) group.

CONCLUSIONSitagliptin can reduce the renal expression of fibronectin by regulating the expression of Egr-1 to achieve renal protection.

Animals ; Apolipoproteins E ; genetics ; Diet, High-Fat ; Dipeptidyl-Peptidase IV Inhibitors ; pharmacology ; Early Growth Response Protein 1 ; metabolism ; Fibronectins ; metabolism ; Gene Knockout Techniques ; Kidney ; metabolism ; Lipids ; blood ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Real-Time Polymerase Chain Reaction ; Sitagliptin Phosphate ; pharmacology

OBJECTIVETo investigate the effect of metformin in protecting against advanced glycation end products (AGEs)-induced apoptosis in human primary dermal fibroblasts.

METHODSFibroblasts were exposed to 100, 200, or 300 µg/mL AGEs, 300 µg/mL bovine serum albumin (BSA), or 300 µg/mL AGEs and 1 mmol/L metformin for 24, 48, or 72 h. The exposed cells were examined for cell apoptosis using a cell counting kit. The expressions of caspase-3, Bax and Bcl-2 protein in the fibroblasts treated for 72 h were detected with Western blotting.

RESULTSAGEs exposures caused significant dose- and time-dependent apoptosis in the fibroblasts. A 72-h exposure to 300 µg/mL AGEs resulted in obviously increased apoptosis of the fibroblasts compared to the control group (0.72 ± 0.02 vs 1 ± 0.04, P<0.05), and metformin significantly decreased AGEs-induced apoptosis (0.98 ± 0.02 vs 0.72 ± 0.02, P<0.05). The expressions of caspase-3 and Bax protein were significantly increased (P<0.05) and Bcl-2 protein expression was decreased (P<0.05) with a lowered Bcl-2/Bax ratio in AGEs-treated fibroblasts (P<0.05), and such changes were significantly reversed by metformin treatment (P<0.05).

CONCLUSIONMetformin can antagonize AGEs-induced apoptosis in human dermal fibroblasts by regulating the expressions of caspase-3, Bax and Bcl-2.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cells, Cultured ; Dermis ; cytology ; Fibroblasts ; cytology ; drug effects ; Glycation End Products, Advanced ; adverse effects ; Humans ; Metformin ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the effect of metformin in protecting against advanced glycation end products (AGEs)-induced apoptosis in human primary dermal fibroblasts. Methods Fibroblasts were exposed to 100, 200, or 300μg/mL AGEs, 300μg/mL bovine serum albumin (BSA), or 300μg/mL AGEs and 1 mmol/L metformin for 24, 48, or 72 h. The exposed cells were examined for cell apoptosis using a cell counting kit. The expressions of caspase-3, Bax and Bcl-2 protein in the fibroblasts treated for 72 h were detected with Western blotting. Results AGEs exposures caused significant dose- and time-dependent apoptosis in the fibroblasts. A 72-h exposure to 300μg/mL AGEs resulted in obviously increased apoptosis of the fibroblasts compared to the control group (0.72 ± 0.02 vs 1 ± 0.04, P<0.05), and metformin significantly decreased AGEs-induced apoptosis (0.98 ± 0.02 vs 0.72 ± 0.02, P<0.05). The expressions of caspase-3 and Bax protein were significantly increased (P<0.05) and Bcl-2 protein expression was decreased (P<0.05) with a lowered Bcl-2/Bax ratio in AGEs-treated fibroblasts (P<0.05), and such changes were significantly reversed by metformin treatment (P<0.05). Conclusion Metformin can antagonize AGEs-induced apoptosis in human dermal fibroblasts by regulating the expressions of caspase-3, Bax and Bcl-2.

Objective To investigate the effect of metformin in protecting against advanced glycation end products (AGEs)-induced apoptosis in human primary dermal fibroblasts. Methods Fibroblasts were exposed to 100, 200, or 300μg/mL AGEs, 300μg/mL bovine serum albumin (BSA), or 300μg/mL AGEs and 1 mmol/L metformin for 24, 48, or 72 h. The exposed cells were examined for cell apoptosis using a cell counting kit. The expressions of caspase-3, Bax and Bcl-2 protein in the fibroblasts treated for 72 h were detected with Western blotting. Results AGEs exposures caused significant dose- and time-dependent apoptosis in the fibroblasts. A 72-h exposure to 300μg/mL AGEs resulted in obviously increased apoptosis of the fibroblasts compared to the control group (0.72 ± 0.02 vs 1 ± 0.04, P<0.05), and metformin significantly decreased AGEs-induced apoptosis (0.98 ± 0.02 vs 0.72 ± 0.02, P<0.05). The expressions of caspase-3 and Bax protein were significantly increased (P<0.05) and Bcl-2 protein expression was decreased (P<0.05) with a lowered Bcl-2/Bax ratio in AGEs-treated fibroblasts (P<0.05), and such changes were significantly reversed by metformin treatment (P<0.05). Conclusion Metformin can antagonize AGEs-induced apoptosis in human dermal fibroblasts by regulating the expressions of caspase-3, Bax and Bcl-2.

Objective:To investigate the frequency and severity of systemic reactions (SRs) to standardized house dust mite subcutaneous immunotherapy (SCIT) in patients with perennial allergic rhinitis (AR), and to analyze the clinical risk factors.Methods:The clinical data of 362 patients including 209 males and 153 females, aged from 5 to 55 years old receiving SCIT at the Department of Otorhinolaryngology, the Third People′s Hospital of Changzhou were collected from May 2014 to July 2017. The SRs were classified as early-onset and delayed-onset, and 4 grades (grade Ⅰ to Ⅳ) to assess severity. The records of SRs were retrospectively analyzed, including the numbers/frequencies, symptoms and signs, onset of reaction and treatment. And the relationships between SRs and patient′s age, gender, allergen injection dose, accompanied allergic diseases were explored. All the statistical analyses were conducted using SPSS 19.0.Results:There were 57 cases (15.75%) of SRs in 362 patients. All the patients received a total of 12 308 injections and 111 SRs (0.90%) were observed. Among them, 31 (27.93%) were early-onset reactions and 80 (72.07%) were delayed-onset reactions; most of the SRs were grade Ⅰ reactions ( n=83, 74.78%), followed by grade Ⅱ ( n=25, 22.52%), grade Ⅲ ( n=3, 2.70%), and no fatal reactions occurred. The incidence of SRs in patients>14 years old was higher than that in patients ≤14 years old according to the number of cases and injections (35.14% vs 13.54%, 2.34% vs 0.76%, χ 2 value was 11.679, 28.162, respectively, all P<0.05), but no significant differences of SRs were observed in gender (18.66% vs 11.76%, 5.98% vs 5.62%, χ 2 value was 3.166, 0.095, respectively, all P>0.05). Fifteen SRs (13.51%) occurred during the build-up phase and 96 (86.49%) during the maintenance phases. SRs could occur in lots of dose phases, and 95 (85.59%) were distributed at high concentrations more than 40 000 SQ-U. The incidence of SRs in patients with multiple allergic diseases was significantly higher than that in patients with AR alone, with asthma or atopic dermatitis (30.67% vs 11.85%, χ 2=15.875, P<0.001). Meanwhile, the incidence of SRs in patients with pure AR was also significantly lower than that in patients with other allergic diseases (5.26% vs 20.56%, χ 2=13.783, P<0.001). Conclusions:The incidence of SRs is less than 1% according to the injection times, the severity of SRs is mostly slight, and the safety and tolerance are good during standardized house dust mite SCIT in perennial AR patients. Delayed-onset SRs are more common. The incidence of SRs is significantly correlated with age, high dose of allergen vaccine injection, and concomitant other allergic diseases (asthma, atopic dermatitis, etc).