The relationship between the characteristics of type 3 secretion system and biofilm of Pseudomonas aeruginosa in diabetic foot wound with antibiotic resistance / 中华内分泌代谢杂志

Objective:To explore the characteristics of type 3 secretion system and biofilm of Pseudomonas aeruginosa in diabetic foot wound, and to analyze the relationship between these factors, as well as to the antibiotic sensitivity.Methods:Thirty-three strains of Pseudomonas aeruginosa were collected from the foot wounds of diabetic foot inpatients in Tianjin Medical University Chu Hsien-I Memorial Hospital from February 1, 2018 to December 31, 2018. Thirteen strains of Pseudomonas aeruginosa were collected from non-diabetic wounds. All strains were tested for antibiotic sensitivity. The virulence genes exoS or exoU of Pseudomonas aeruginosa and the ability of biofilm formation were tested. The characteristics of exoS or exoU and biofilm of Pseudomonas aeruginosa were analyzed. Patients′ clinical outcomes were also analyzed.Results:Pseudomonas aeruginosa with exoS gene was the major pathogen, 90.9% found in diabetic foot group and 84.6% in control group, with no significant difference( χ2=0.54, P=0.46). The drug-resistant strains of Pseudomonas aeruginosa with exoS accounted for 16.7% in diabetic foot group and 18.2% in control group, also with no significant difference( χ2=0.18, P=0.83). There were 5 strains of Pseudomonas aeruginosa carrying exoU, 3 strains in diabetic foot group, of which 1 was resistant, 2 in control group, no resistant strain. Pseudomonas aeruginosa increased the ability of biofilm formation in diabetic foot group, accounting for 57.6%, and for resistant strains, 83.3% of them increased the biofilm formation ability. Two kinds of Pseudomonas aeruginosa produced different biofilms, but they were effectiveless for carbapenem antibiotics. The times of debridement ( P<0.01), time of antibiotic use ( P<0.01) were more in biofilm wound, but the healing rate reached 75%-90%. Conclusion:Pseudomonas aeruginosa secreting ExoS is the main one in the diabetic foot wound. The ability of Pseudomonas aeruginosa to produce biofilm in DF wound is increased. Biofilm is one reason for its antibiotic resistance. Multiple debridement combined with sensitive antibiotics is an effective method to remove biofilm.

Effects of salvia miltiorrhiza polyphenols for injection and collagen sponge in diabetic foot / 中国生化药物杂志

Objective To investigate the effect of polyphenol of salvia miltiorrhiza polyphenols combined with collagen sponge in the treatment of diabetic foot.Methods A total of 115 patients with type 2 diabetic foot were randomly divided into treatment group (55 cases) and control group (60 cases).Salvia miltiorrhiza polyphenols and collagen sponge treatment group were treated with conventional treatment plus type 2 diabetes treatment of salvia miltiorrhiza polyphenol combined with collagen sponge;4 weeks as a treatment course, observation of 8 weeks, ulcer and other adverse reactions were recorded and compared.Results After 4 weeks of treatment, 17 cases (30.91%) with 30% or more reduction in foot ulcer area in treatment group,33 cases (60.00%) after 6 weeks treatment and the number of cases with 30% or more reduction in foot ulcer area,49 cases (89.09%) after 8 weeks of treatment, and the number of cases and the proportion of cases with 30% or more decrease of ulcer area in the control group after 4, 6 and 8 weeks were 12(20.00%), 22 (36.67%) and 39 (65.00%).There was no statistical difference between the two groups for the healing time of foot ulcer at three time points.After 8 weeks of treatment, the cure rate was 38.18% in salvia miltiorrhiza polyphenol combined with collagen sponge group. The effective rate was 50.91% and 45.00% for the two groups, the difference was not statistically significant; while the total effective rate was also significantly higher(89.09% vs.65.00%,P<0.01).Conclusion Salvia miltiorrhiza polyphenol combined with collagen sponge has a good effect on diabetic foot treatment, especially it can improve the cure rate, which is superior to the routine therapy.

Glucagon-like peptide 1 receptor agonist protects high-glucose inducedβcells apoptosis via inhibition of NOX2-dependent ROS production / 天津医药

Objective To investigate the possible mechanisms of glucagon-like peptide 1 receptor agonists (GLP-1Ra) protection against hyperglycemic induced beta cell apoptosis through depression of NOX2-dependent ROS production. Methods The rat model of type 2 diabetes (T2DM) was established by injecting small doses of streptozotocin (STZ) fol?lowed by 8-week high fat diet. The experimental animals were divided into three groups:normal control (N) group, diabetes (T2DM) group and GLP-1Ra group [treated with liraglutide 200 μg/(kg · d)for 12 weeks]. The blood glucose levels were compared before and after modeling, before treatment and 12-week after treatment with GLP-1Ra. The level of glycosylated hemoglobin (HbA1c) was detected by high-pressure liquid chromatography. Automatic biochemical analyzer was used to de?tect levels of aspertate aminotransferase (AST), creatinine (CR) and urea nitrogen (BUN). The apoptotic rates of islets were determined by TUNEL method and cleaved caspase 3 was detected by immunohistochemistry. DCFH-DA fluorescent probe was used to detect reactive oxygen species (ROS) levels of islets. Levels of NADPH oxidase (NOX) catalytic subunit (NOX 2) in islets were measured by immunohistochemistry. Results At the end of the study, glycemic control (average blood glucose/week and HbA1c) and lipid situation were improved significantly in the GLP-1Ra group than those of N group (P0.05). After application Apocynin for inhibition, there were no significant differences between three groups (P>0.05). The level of NOX2 was significantly lower in GLP-1Ra group compared to that of T2DM group (P<0.05). Conclusion GLP-1Ra can inhibit apoptosis ofβcells in diabetes rat, and the depression of NOX2-dependent ROS may be one of the important underly?ing mechanisms.

Effect of high glucose and anoxia on Amot expression in vascular endothelial cells with regard to its function in promoting angiogenesis / 中华内分泌代谢杂志

Objective To observe the effects of high glucose and anoxia on Amot expression in vascular endothelial cells (VECs),and explore its role in angiogenesis.Methods VECs were incubated with different glucose concentrations for 48 h,and then cultured at normal oxygen concentration or anaerobic condition for 24 h.The protein expressions of p130-Amot and p80-Amot were detected by Western blot.After Amot expression was downregulated in VECs by siRNA,wound healing experiments and angiogenesis experiments were performed to test the effect of decreased Amot expression on angiogenesis.Results pl30-Amot protein expressions in low glucose (5.5mmol/L) plus normal oxygen group and low glucose plus anaerobic group were higher than those in high glucose (30mmol/L) plus normal oxygen group,high glucose plus anaerobic group,middle glucose (15 mmol/L) plus normal oxygen group,and middle glucose plus anaerobic group (all P＜0.01).Compared with low glucose plus anaerobic group,p130-Amot expression was higher in low glucose plus normal oxygen group (P ＜ 0.01).However,the expression of p80-Amot showed no statistically significant difference among different groups (P＞0.05).Compared to the normal VECs,the cells with decreased Amot expression by siRNA exhibited an attenuated cell migration in the wound healing experiments and a lesser tube formation in the angiogenesis experiments.Conclusions High glucose exerts a more significantly negtive effect on the Amot expression than anoxia in VECs.The downregulation of Amot expression inhibits migration and angiogenesis of VECs.

Study on the safety and effectiveness of an exercise prescription for patients with diabetic foot ulcer and peripheral arterial disease / 中华内分泌代谢杂志

Objective To investigate the effect of exercise prescription on foot perfusion and ulcer healing in patients with diabetic foot ulcer (DFU) accompanied with peripheral arterial disease (PAD).Methods Sixty patients with DFU and PAD were divided into exercise group and control group.The patients in the exercise group took exercise therapy training,ensuring that they could follow the exercise prescription.The patients in the control group had no exercise requirements.After the treatment for 12 weeks,ankle brachial index (ABI),transcutaneous oxygen tension (TcPO2),skin temperature,body mass index (BMI),HbA1C,and the ulcer healing rate in the two groups were compared,the satisfaction and compliance in the exercise group were evaluated,and the adverse events of the treatment were recorded.Results After the treatment for 12 weeks,the ABI,TcPO2,and foot skin temperature in the two groups increased and HbA1C decreased(P＜0.05 or P＜0.01).TcPO2 increased 5.25 mm Hg(1 mm Hg=0.133kPa),skin temperature increased 0.45℃,and BMI decreased 0.69 kg/m2 in the exercise group,while TcPO2 increased 2.59 mm Hg,skin temperature increased 0.28℃,and BMI increased 0.02 kg/m2 in the control group,showing significant differences in the three index changes between two groups (P＜0.01).The healing rate in the exercise group was higher than that in the control group (53.6％ vs 25.9％,P ＜0.05).The patients'compliance and satisfaction to the exercise therapy were 90％ and 94％,respectively.The incidence of adverse events in the exercise group had no causal relationship with the exercise prescription.Conclusion The exercise prescription in the study can improve the foot perfusion,and promote ulcer healing.It has the advantage of high safety,patients'compliance and satisfaction.

Clinical Features and Antibiotic Sensitivity of Gram-Negative Diabetic Foot Osteomyelitis / 天津医药

Objective To investigate the clinical features and antibiotic susceptibility of osteomyelitis infected by Gram-negative bacteria (G-) in patients suffered from diabetic foot ulcers (DFU). Methods The clinical data of 91 DFU pa-tients accompanied with osteomyelitis (DFO) were retrospective studied. These patients hospitalized in the Tianjin Metabolic Diseases Hospital were divided into two groups, Gram-negative bacteria (G-) group (n=44) and Gram-positive bacteria (G+) group (n=42), respectively. The clinical features were compared between two groups. Logistic regression analysis was used to determine the risk factors for Gram-negative bactreial infection. The Gram-negative antibiogram was summarized. Results A total of 112 pathogens were isolated from 91 patients. G-bacteria were the most frequent pathogens (48.2%), following by G+ bacteria (47.3%) and fungi (4.5%). Pseudomonas aeruginosa was the majority of the G-bacteria. Comparing the two groups, the rate of antibiotic use within the previous 6 months was significantly higher in G-group (75.0%) than that of G+group (52.4%, P<0.05). There were no significant differences in the other indicators between two groups. The Logistic re-gression analysis revealed that the history of antibiotic use was the independent risk factor of G-bacterial infections in DFO patients. Antibiotics susceptibilities reflected G- bacteria were more prevalent to resist to cephalosporins and quinolonem, but sensitive to imipenem, ceftazidine and cefperazone-sulbactam. Conclusion Gram negative bacteria were not only the main pathogens isolated from DFO patients, but also frequently resistant to several popular antibiotics in China. The proper bacteria culture and antibiotic sensitivity test are especially emphasized to patients with DFU.

The baseline characteristics of patients with type 2 diabetes initiating insulin detemir:the Chinese cohort from the SOLVETM study / 中华内科杂志

Objective To characterize the baseline status of Chinese diabetic patients based on data derived from Chinese cohort from SOLVETM study.Methods Patients with type 2 diabetes initiating basal insulin detemir at the decision of the physician were eligible for the study.Data on demographics,medical history,glycemic profile and treatment regimen at baseline were collected by physicians.Results A total of 3272 patients [female 42％,male 58％,mean age (56.2 ± 10.8) years] were included in the study.Their BMI was (25.3 ± 3.3) kg/m2.The duration of diabetes was 4.0 (0.1-27.0) years,and the duration of treatment with oral antidiabetic drugs (OADs) was 3.0(0.0-20.2) years.The proportions of subjects with diabetic macro-and micro-vascular complications were 15.8％ (515 cases) and 27.1％ (866 cases),respectively.The hemoglobin Al c (HbAl c) at baseline was (8.33 ± 1.70) ％,and the fasting blood glucose (FPG) was (9.5 ± 2.6) mmol/L.Conclusions A large proportion of patients with type 2 diabetes remain in poor glycemic control,and the prevalence of diabetic complications is high,which requires optimal therapeutic strategy for the patients with suboptimal glycemic control.

Infection of multidrug-resistant Pseudomonas aeruginosa and its risk factors in diabetic foot / 中华临床感染病杂志

Objective To investigate the infection and risk factors of multidrug-resistant Pseudomonas aeruginosa in diabetic foot.Methods Totally 85 strains of Pseudomonas aeruginosa were isolated from 428 samples of diabetic foot ulcers in Tianjin Metabolic Diseases Hospital from Jan 2008 to Dec 2010.Drug sensitivity tests were performed and multivariate logistic regression analysis was used to examine the risk factors of multidrug-resistant Pseudomonas aeruginosa infection. Results In 85 strains of Pseudomonas aeruginosa, 28 (32.94％ ) were multidrug resistant. Multidrug-resistant Pseudomonas aeruginosa showed high resistance to 3rd generation cephalosporins,quinolones and aminoglycosides with resistant rates of 42.86％-67.86％,32.14％ -57.57％,and 42.86％-67.86％,respectively. Logistic analysis indicated that previous antibiotic treatments, hypertension and anemia were associated with multidrug-resistant Pseudomonas aeruginosa infection ( OR =5.758,0.257 and 0.270,P =0.006,0.014 and 0.013 ). Conclusion Multidrug-resistant Pseudomonas aeruginosas isolated from diabetic foot are highly resistant to several commonly used antibiotics,and previous antibiotics use,hypertension and anemia are risk factors for multidrug-resistant Pseudomonas aeruginosa infections.

Clinical features and drug resistance of pseudomonas aeruginosa isolates from patients with diabetic foot infections / 中华内分泌代谢杂志

Objective To investigate clinical features and antibiotic resistance of pseudomonas aeruginosa (PA) strains isolated from patients with diabetic foot infections (DFI) in Tianjin Metabolic Diseases Hospital.Methods Eighty-five PA strains were isolated from 428 patients with diabetic foot in the hospital from Jan 2008 to Dec 2010.The clinical features of patients were summarized.Relationships between the isolates and depth of ulcer or severity of infection were analyzed.The disk-diffusion method was performed to examine antimicrobial susceptibility.Results Gram positive (G+) and Gram negative (Gˉ) isolates were 50.47％ and 41.12％,respectively.Multidrug-resistant PA composed 32.9％ of the total PA isolates.The size of ulcers with PA infections was bigger than those with non-PA bacterial infections (P＜0.05).Compared to G+ strains,patients with PA strains were older,had lower hemoglobin,but higher serum sensitive C-reactive protein; and more frequently,they had ischemic ulcer and osteomyelitis.Compared to G+ strains,the PA strains were more frequently isolated from deeper ulcers and with more serious infections(P＜0.05).The resistant rates of PA to cephalosporins,fluoroquinolones,and aminoglycosides were between 32.9％-61.2％,37.6％-42.4％,and 37.6％-62.4％,respectively.Only one out of 85 PA strains was imipenem-resistant.However,sensitiveness of all PA isolates to cefoperazone and sulbactam reached 100％.Conclusion PA strains are mainly found in patients with deeper ulcers and more serious infections.Multidrug-resistant PA is common in DFI.It is important to isolate pathogens and determine their antibiotic resistance correctly in diabetic foot patients in order to provide appropriate drug administration and to reduce the production and dissemination of drug resistant strains.

The outcomes of toe amputations in diabetic patients: a 5-year follow-up study / 中华内分泌代谢杂志

Objective To observe the outcomes of toe amputations in diabetic patients and to analyze the affecting factors.Methods Two-hundred and forty-five patients were divided into 2 groups:202 cases with the wound healed after toe amputation,and 43 patients whose wound did not heal after toe amputation.The factors related to healing were analyzed.Patients with healed wounds after toe amputations ( n =202 ) were followed,regarding reappearance of new ulcers,reamputations,mortality,activities of daily life,and the affecting factors.Results The rate of wound healing was 82.4％.The rate of follow-up in the healed group was 91.6％.The rates of reappearance of new ulcers after 1,3,and 5 years of observation were 27.3％,57.2％,76.4％,respectively.The rates of reamputations after 1,3,and 5 years of observation were 17.5％,22.3％,47.1％,respectively.The mortality rates by 1,3,and 5 years after the toe amputations were 3.8％,15.1％,32.7％,respectively.Mean survival time of the fatal cases after the first amputation was 3.8 (95％ CI 3.4-4.1 )years.Multivariate analysis showed that HbA1c (＞9％)was an independent predictor of wound healing,reappearance of new ulcers,and reamputations after toe amputations,and age ( ＞ 70 years ) was an independent predictor of reamputations,mortality,and hampered daily activities.Conclusion Despite a satisfactory initial healing process after the first amputation,the long-term outcomes of the followed-up patients are not optimistic.Senile patients are often with poor prognosis,and good blood glucose control may improve the prognosis.

SCCmec genotypes of methicillin-resistant Staphylococcus epidermidis in diabetic foot infections / 中华微生物学和免疫学杂志

Objective To investigate SCCmec genotypes and drug-resistance profiles of the methieillin-resistant Staphylococcus epidermidis (MRSE) strains isolated from the patients suffered from diabetic foot infections (DFI) in the Tianjin Metabohc Diseases Hospital. Methods After dabridement, specimens of 390 infectious diabetic foot ulcers in the hospital from Jan 2008 to Jun 2010 were collected from the wound basal parts by cotton swab for culture. The disk-diffusion method was performed to examine antimicrobial susceptibility. DNAs of the MRSE strains were extracted, and their SCCmec genotypes were identified by PCR. Results Twenty of the seventy(28.6% ,20/70)Staphylococcus epidermidis strains were mecA posifive. Among the MRSE isolates, 2 ( 10.0% )were SCCmec Ⅱ ,9 (45.0%)were SCCmecⅢ and 9 (45.0%)were SCCmec Ⅳ. None of the isolates were genotyped as SCCmec Ⅰ or Ⅴ. No mater which genotypes they were, all the MRSE isolates were multi-drug resistant. They were resistant not only to β-lactams (including penicillins, cefoxitin and cephems), but also to non-β-lactams (including macrolides, fiuoroquinolones and sulfonamides ) . Resistance to voncomycin and rifampicin were not found in these strains . Conclusion SCCmec Ⅲ and SCCmecⅣ are major genotypes of the MRSE isolates from the infectious diabetic foot ulcers.The SCCmec Ⅳ genotype strains with multi-drug resistant profiles are prevalent in the diabetic foot infections.

Reseach on epidermal keratinocyte function impairing in rats with diabetes mellitus / 中华老年医学杂志

Objective To explore the effect of diabetes mellitus (DM) on biological behavior of epidermal keratinocyte in rats. Methods A total of 40 Sprague-Dawley rats were randomized into control group and streptozotocin (STZ) -induced diabetes group. Of each group, 10 rats were implemented with deep partial-thickness scalding. The re-epithelialization rate was observed at the 3rd, 7th, 14th and 21th post-burn day. Histological characteristics and thickness of epidermal tissue in both groups were observed. The adhesion rate, cell cycles, apoptosis rate and migration ability of keratinocyte were measured. The accumulation of advanced glycosylation end products (AGEs) of epidermal tissue was observed. Results The percentages of re-epithelialized area at the 7th, 14th and 21th post-burn day were much lower in DM group than in control group (P<0.05). In DM group, the epidermal thickness was reduced obviously with obscure multilayered epithelium and less amount of prickle cells; The adhesion rates of 12, 24 h after culturing keratinocyte and the percentage of G2/M phase cells were lower in DM group than in control group (P<0.05). However, apoptosis rate of keratinocyte was higher, the amount of migration cell was significantly less in DM group than in control group (both P<0.05). There were lots of AGEs accumulated in epidermal tissue in DM group, while there were hardly AGEs in control group. Conclusions Re-epithelization blocked exists on non-healing wound in DM rats, which is related with the impaired keratinocyte biological behavior. A large of AGEs accumulate in the epidermal tissue of DM rats, which may be a important reason to inhibit keratinocyte function in diabetic environment.

Expression and significance of caveolin-1 in femoral nerve of diabetic foot amputation patients / 中华内分泌代谢杂志

Objective To investigate the expression and significance of caveolin-1 in femoral nerve of diabetic patients with foot amputation. Methods Forty patients with foot amputation were assigned to 3 groups according to their duration of type 2 diabetes: group A ( ＜6 years＝, group B (6-10 years), and group C ( ＞10 years). Hematoxylin and eosin (HE) stain and Weil's stain were used to examine the femoral nerve. Silver staining was used to observe the axons and to count the nerve fiber density. The expression of caveolin-1 in Schwann cells of femoral nerve was tested by immunohistochemisty. Results There were evident progressive pathological changes in femoral nerve in the 3 groups. The variance of nerve fiber density in the 3 groups reached statistical significance ( P＜0. 05 ＝, the nerve fiber density showed negative correlation with HbA1C( r =-0. 792, P＜0. 01 ＝ and duration ( r=-0.592, P＜0. 01 ＝. The expression of caveolin-1 in Schwann cells of femoral nerve was positive in all the 3 groups and the variance with statistical significance (P＜0. 01 ), it was negatively correlated with HbA1C (r=-0. 762, P＜0. 01 )and duration (r=-0. 532, P＜0. 01 ), and it was positively correlated with nerve fiber density (r=0. 721, P＜0.01 ), the partial correlation coefficient of caveolin-1 and HbA1Cwas-0. 505 ( P＜0. 01 ).Conclusion In patients with diabetic foot amputation, caveolin-1 may play a role in the development of diabetic peripheral neuropathy and diabetic foot.

Repair of ulcer with rhEGF sustained-release microspheres in diabetic rats / 中华创伤杂志

Objective To prepare recombinant human epidermal growth factor (rhEGF) sustained-release microspheres and evaluate their morphology, rhEGF releasing activities and cell proliferation activity in vitro and compare difference of rhEGF sustained-release microspheres and rhEGF in facilitaring ulcer healing in diabetic rats. Methods (1) rhEGF sustained-release microspheres were prepared by the modified double emulsion method. Morphology of the microspheres was detected by transmission electron microscope and size distribution measured by laser granularity meter/Zeta electric potential meter. ELISA assays were applied to determine rhEGF releasing. (2)Proliferation of mouse fibroblasts was analyzed by MTr method. (3) Diabetic rat models were prepared and divided into four groups, ie, rhEGF sustained-release mierospheres group (Group A), rhEGF stock solution group (Group B), blank sustainedrelease mierospheres group (Group C) and PBS meustruum control group (Group D), which were given drug once a day. The wound healing rate was calculated by taking photographs at days 3,7,14 and 21. Skin specimens from the wound edge were harvested partially for observation of hydroxyproline (HYP) contents. Immunohistochemistry was employed to detect integrin 131 and keratin-19 and measure their positive staining area ratio. Results (1) The particle diameter of rhEGF sustained-release microspheres was 193.5 nm, with relative uniform particle diameter distribution. There showed no conglutination among rhEGF susrained-release microspheres, with good dispersibility. Releasing drug lasted for 24 hours and accorded with Higuchi release kinetic model. (2) Different concentrations of rhEGF sustained-release microspheres could promote the proliferation of mouse fibroblast, especially the concentration of 10 μg/L (P <0.05, compared with the control). (3) From the 7th day after treatment, Group A had the fastest wound healing rate, with statistical difference compared with other three groups (P < 0.05). Group A had higher HYP contents and positive area ratio of integrin β1 and keratin-19 than Group B. Conclusions rhEGF sustained-release microspheres prepared by the modified double emulsion method have uniform particle size and can last release for 24 hours. Compared with rhEGF stock solution, rhEGF sustained-release microspheres have faster and better ulcer healing and higher healing quality in diabetic rats.

Effects of Pharmaceuticals Pollution in the Aquatic Environment on Aquatic Organism and Human Health / 环境与健康杂志

The pharmaceuticals have been playing an important role in insuring human health and stockbreeding for decades. However incompletely absorbed pharmaceuticals will find their way into the wastewater in their original structure or metabolites. The ineffective operation of wastewater-treatment plants makes it possible that the widespread appearance of pharmaceuticals in the aquatic environment. The pharmaceuticals become the kind of "persistent" pollutants in the aquatic environment since they have been widely and continually used in daily life although their half-lives are relatively short. Their occurrences threaten the aquatic ecological safety and human’s health. In recent years, European and North American countries have focused on the research of pharmaceuticals in the aquatic environment, while the related reports are scarce in China. This article comprehensively analyzed and discussed the effects of pharmaceuticals pollution in the aquatic environment on aquatic organism and human health. It will present an important reference for future related research as well as establishing ecological risk assessment of pharmaceuticals in aquatic environment in China.