[Pathogen analysis in patients with diabetic foot osteomyelitis using 16S rRNA high-throughput sequencing].

OBJECTIVE: To analyze the characteristics of pathogenic microorganisms in the infected bone tissues in patients with diabetic foot osteomyelitis (DFO) using 16S rRNA high-throughput sequencing to facilitate rapid and accurate detection of pathogens and effective infection control. METHODS: Between September, 2016 and April, 2017, 16 patients with DFO were admitted in our department and infected bone specimens were obtained during debridement. The pathogenic microorganisms in the specimens were identified using both 16S rRNA high-throughput sequencing and automatic blood culture analyzer, and the characteristics of the microflora were analyzed based on 16S rRNA sequencing data in comparison with the results of blood culture. RESULTS: The results of 16S rRNA sequencing showed that bone tissues of DFO contained diverse and uniformly distributed pathogenic organisms, among which 20 (87%) dominant genera were identified with Prevotella as the most abundant pathogen. Both 16S rRNA sequencing and routine culture results suggested the domination of gram-negative bacteria among the pathogens in DFO bone tissues. 16S rRNA sequencing, compared with routine culture, yielded a higher positivity rate (100% vs 88.24%) and detected a greater average number of pathogens (12.56 vs 1.50) and a higher proportion of gram-negative bacteria (67.16% vs 50.00%) in the samples. 16S rRNA sequencing detected nearly all the pathogens identified by routine culture except for Escherichia coli, Serratia marcescens and Enterobacter cloaca, and identified 13 genera that failed to be detected by routine culture, including the obligate or strict anaerobes Anaerococcus, Veillonella, Bacteroides, Fusobacterium, Porphyromonas, Finegoldia, Prevotella, Peptostreptococcus, Parvimonas, Peptoniphilus and Bulleidia. Routine culture did not detect any anaerobes in the samples but identified multidrug-resistant strains in as many as 58.33% of the pathogens. CONCLUSIONS: 16S rRNA high-throughput sequencing is capable of demonstrating the diversity and abundance of microflora in DFO bone tissues, where diverse and uniformly distributed pathogens can be detected with a discrete distribution of the dominant genera, most of which are gram-negative. Compared with routine culture method, 16S rRNA sequencing allows more convenient and accurate identification of the pathogens (especially gram-negative bacteria and anaerobes), and can be useful in clinical decision on appropriate treatment of DFO.

[Anlysis of foot biomechanics characteristic in 303 patients with type 2 diabetes mellitus].

OBJECTIVE: To investigate foot biomechanics characteristic of patients with type 2 diabetes mellitus. METHODS: This study was conducted among 303 patients with type 2 diabetes. The whole foot was divided into 10 regions, namely the first toe (T1); the second to fifth toes (T2-5); the first, second, third, fourth, and fifth metatarsals (M1, M2, M3, M4, and M5, respectively); midfoot (MF), and the heel medial (HM). Foot arch index, foot angle and maximum peak pressure (MPP) of the 10 regions were measured using a Footscan gait system. RESULTS: The maximum peak pressure of 10 regions decreased in the order of M3>M2>HM>M4>HL>M1>M5>T1>ML>T2-5 for the left foot, and in the order of M3>M2>HM>M4>HL>M1>M5>T1>ML>T2-5 for the right foot. The MPP in M1 region was higher in the right than in the left foot (P<0.05). The MPP in M3, M4, M5, and MF was higher in the left than in the right foot (P<0.05). The percentage of high-risk foot (defined by a total plantar pressure ≥70 N/cm2) was 34% on the left and 17.7% on the right. An increased BMI was associated with a significant increase in high-risk foot, but not for the right foot in underweight patients. Foot flat phase was extended and forefoot push-off phase shortened in stance phase in the patients. Compared with the right foot, the left foot showed a significantly increased foot arch index and increased low and high arch rates with a decreased normal arch rate. Total plantar pressure was higher in of the left high arch foot than in normal arch foot. The foot angle was significantly larger on the right than on the left. The bilateral total plantar pressures were significantly greater in male patients (P<0.05) and increased with age but were not associated with the duration of DM, foot angle, or glycosylated hemoglobin level. CONCLUSION: Diabetic patients have obvious alterations in foot biomechanics with abnormalities of the plantar pressure, and the percentage of high-risk foot increases in overweight and obese patients, suggesting the need of body weight control in these patients when administering offloading treatment for prevention of diabetic foot ulcer.

ERα inhibits epithelial-mesenchymal transition by suppressing Bmi1 in breast cancer.

In human breast cancer, estrogen receptor-α (ERα) suppresses epithelial-mesenchymal transition (EMT) and stemness, two crucial parameters for tumor metastasis; however, the underlying mechanism by which ERα regulates these two processes remains largely unknown. Bmi1, the polycomb group protein B lymphoma Mo-MLV insertion region 1 homolog, regulates EMT transition, maintains the self-renewal capacity of stem cells, and is frequently overexpressed in human cancers. In the present study, ERα upregulated the expression of the epithelial marker, E-cadherin, in breast cancer cells through the transcriptional down-regulation of Bmi1. Furthermore, ERα overexpression suppressed the migration, invasion, and EMT of breast cancer cells. Notably, overexpression of ERα significantly decreased the CD44high/CD24low cell population and inhibited the capacity for mammosphere formation in ERα-negative breast cancer cells. In addition, overexpression of Bmi1 attenuated the ERα-mediated suppression of EMT and cell stemness. Immunohistochemistry revealed an inverse association of ERα and Bmi1 expression in human breast cancer tissue. Taken together, our findings suggest that ERα inhibits EMT and stemness through the downregulation of Bmi1.

Interferon alpha on expression of hTERT mRNA in peripheral blood mononuclear cells of patients with chronic hepatitis B.

Cell division is closely related to telomerase activity (hTERT mRNA). Lower expression of lymphocitic hTERT mRNA may easily cause cell aging, which is not beneficial to maintaining a durable lymphocyte division. To date, there is no study to investigate IFNα therapy on hTERT mRNA expression in PBMCs of patients with chronic hepatitis B (CHB). We quantitatively detected hTERT mRNA from study subjects and made each hTERT mRNA normalized (NhTERT mRNA). Mean NhTERT mRNA level was lower in either CHB group, but it significantly increased in IFNα-treated group compared with CHB control group, and a longer duration of IFNα therapy could increase the level. Moreover, the mean NhTERT mRNA in subgroup with HBeAg loss was significantly higher than that in subgroup without. NhTERT mRNA was markedly correlated with CD3(+) T lymphocyte count and CD4(+)/CD8(+) ratio. The results showed that IFNα therapy could upregulate the expression of hTERT mRNA in PBMCs.

[Efficacy and safety evaluation of two insulin treatment protocols using a continuous glucose monitoring system].

OBJECTIVE: To evaluate the efficacy and safety of two insulin treatment protocols using a continuous glucose monitoring system. METHODS: Type 2 diabetic patients mellitus with unsatisfactory control of fasting blood glucose by oral antidiabetic drugs were included in the study. The patients were randomized into two groups to receive bedtime injection of glargine and oral antidiabetic drugs (group A) or injection of Novolin 30 R twice a day (group B) for 12 weeks. The insuline dose was adjusted according to fasting blood glucose till discharge. Continuous glucose monitoring system was used to record the average blood glucose, fasting blood glucose, 2 h postprandial blood glucose, AUCPG ≥ 10.0 mmol/L%, HbA1c and C peptide, bedtime blood glucose, 3:00 AM blood glucose, the incidence of hypoglycemia and body mass index. RESULTS: The average blood glucose, fasting blood glucose, 2 h postprandial blood glucose, AUCPG ≥ 10.0 mmol/L% and HbA1c was significantly decreased and C peptide significantly increased in the two groups after the treatments. The patients in glargine group showed better improvement with a significantly lower incidence of hypoglycemia than those in Novolin 30 R group. BMI underwent no significant changes in the two groups after the treatments. CONCLUSION: Glargine therapy better mimics the physiological insulin secretion patterns, and when combined with oral antidiabetic drugs, can be more effective and safer than premixed insulin.

[Epidemiological investigation of diabetes and prediabetes in community residents in the suburbs of Guangzhou].

OBJECTIVE: To identify the prevalence and risk factors of diabetes and prediabetes in the community residents above 18 years old in the suburbs of Guangzhou. METHODS: Between April and May in 2008, the residents above 18 years living in 6 communities of Guangzhou for 5 or more years were sampled with multistage clustering sampling. The sampled residents were surveyed by questionnaires, and physical examination and glucose determination were carried out. RESULTS: A total of 1532 residents were sampled. The incidence of diabetes mellitus in these community residents was 8.46%, and that of impaired glucose regulation was 6.59%. Age, body mass index, family history of diabetes mellitus, case history of hypertension, hyperlipidemia, hypertension and smoking were all the independent risk factors for impaired glucose regulation and diabetes mellitus. CONCLUSION: Diabetes and prediabetes are prevalent in the community residents in Guangzhou. Controlling the risk factors such as obesity, hypertension, lipid metabolism disorder among the residents above 40 years with a family history of diabetes mellitus and hypertension is key to prevention of impaired glucose regulation and diabetes mellitus.

[Association of blood glucose fluctuation with the pain sensation in patients with painful diabetic neuropathy].

OBJECTIVE: To investigate the relationship between blood glucose fluctuation and the pain sensation in patients with painful diabetic neuropathy. METHODS: The severity of pain in 16 patients with painful diabetic neuropathy was assessed using visual analog scale (VAS), and according to the results, the patients were divided into mild to moderate pain group and severe pain group. The blood glucose of the patients were monitored continuously for 72 h using a continuous glucose monitoring system, and the MBG, SD, DMMG, MAGE, and AUCPG were obtained with the FBG and HbA1C tested. RESULTS: The SD, DMMG, and MAGE of the severe pain group were all significantly higher than those of the mild to moderate pain group (P<0.05), and no significant difference was found in MBG, FBG or HbA1c between the two groups. CONCLUSION: The blood glucose fluctuation is associated with the severity of the pain degree, and effective control of the blood glucose fluctuation helps prevent and treat painful diabetic neuropathy.

[Antibiotic susceptibility of pathogenic bacteria isolated from 893 children with lower respiratory infection in Guiyang].

OBJECTIVE: To investigate the distribution and the antibiotic susceptibility of pathogenic bacteria in children from Guiyang with lower respiratory infection (LRI). METHODS: The nasopharyngeal aspirate samples were obtained from 893 hospitalized children with LRI between August 2006 and June 2008. An antibiotic susceptibility test was performed using the VITEK system and the Kirby-Bauer diffuse method after bacteria were identified. RESULTS: Five hundred and forty-three patients (60.8%) were bacteria-positive. A total of 598 strains (30 kinds of bacteria) were obtained from the sputum samples. Of them, 533 strains (89.1%) were gram-negative and 57 were gram-positive (9.8%). Escherichia coli (E. coli) and Kleb-siella pneumoniae (K. pneumoniae) were common in gram-negative strains. They were susceptive to piperacillin/tazobactam, amikacin, ciprofloxacin, and levofloxacin, especially to imipenem. Streptococcus pneumoniae (S. pneumoniae) and Stapthylococcus aureus (S. aureus) were common in gram-positive strains. S. pneumoniae was susceptive to penicillin and cefazolin sodium, but S. aureus was resistant. Both were high susceptive to vancomycin, and resistant to roxithromycin. CONCLUSIONS: Gram-negative bacteria are the main pathogens in children from Guiyang with LRI, and E. coli and K. pneumoniae are common. The antibiotic susceptibility of pathogenic bacteria varies with different strains of bacteria. A reasonable selection of antibiotics should be based on the antibiotic susceptibility test.

[Cloning and secretory expression of islet neogenesis-associated protein in Pichia pastoris].

OBJECTIVE: To clone the recombinant human islet neogenesis-associated protein (rhINGAP) gene for its secretory expression in Pichia pastoris. METHODS: INGAP gene was amplified with PCR and inserted between Xho I and EcoR I downstream sites of the alpha factor of the recombinant plasmid alpha/pUC18. The fusion gene of alpha factor and INGAP was subsequently inserted between BamH I and EcoR I sites of the plasmid pPIC9K of P. pastoris. After confirmation with restriction enzyme digestion and sequencing, the positive recombinant plasmid that integrated INGAP gene was linearized with Sal I digestion and transformed into the yeast host strain GS115 through electroporation. The yeast transformants that harbored the INGAP gene with high copies were selected with the auxotroph medium and G418, followed then by PCR verification of the positive transformants, from which the expression of recombinant human INGAP was induced with methanol as the only carbone source. The antigenic activity of the desired protein was then detected using Western blotting and enzyme-linked immunosorbent assay (ELISA). RESULTS AND CONCLUSION: The recombinant expression plasmid INGAP/pPIC9K was successfully constructed, and 3 positive transformants were obtained. The expressed protein showed good antigenic activity as confirmed by Western blotting and ELISA.

[Sweat function evaluation for early diagnosis of diabetic peripheral neuropathy].

OBJECTIVE: To evaluate the clinical value of sweat function examination in early diagnosis of diabetic peripheral neuropathy (DPN). METHODS: Ninety-eight hospitalized type 2 diabetic patients with or without DPN (DN and DC groups) according to Michigan Diabetic Neruopathy Score (DNS) and 40 healthy volunteers (NC group) were evaluated for their sweat function of the feet in relation to the peripheral autonomic nerve with sweat printing method using Neuropad. The Neuropad color-changing time was recorded to assess the sensitivity and specificity of sweat printing methods relative to DNS for DNP evaluation, and the correlation of the Neuropad color-changing time to DNS score was analyzed. RESULTS: The average Neuropad color-changing time was 4.0-/+0.6, 4.3-/+1.2 and 23.0-/+6.1 min in NC, DC, and DN groups, respectively, showing significant differences between the 3 groups (P<0.05). The morbidity rate detected by sweat printing method was 62.2%, similar to that detected by DNS (57.1%, P>0.05). The sensitivity of the sweat printing method for DPN diagnosis was 92.8%, with specificity of 78.5%, positive predictive value of 93.2%, and negative predictive value of 78.6%. DNS showed significant positive correlation with the Neuropad color-changing time (r=0.46, P<0.05). CONCLUSION: Sweat printing method provides an objective, simple and reliable method for sweat function evaluation of the feet of type 2 diabetic patients to help in early DPN diagnosis, and quantification of the results of sweat printing method can be indicative of the DPN severity.

[Prevalence of microalbuminuria in type 2 diabetic patients with hypertension].

OBJECTIVE: To investigate the prevalence of microalbuminuria in type 2 diabetic patients with hypertension. METHOD: Urinary albumin excretion was measured in 55 diabetic patients with hypertension by colorimetric methods using commercially available test strips of Nephur 7 and Micro-II, the latter test strips employed only when the former yielded negative results for proteinuria. The height, body weight, waist circumference and blood pressure were measured for all the patients with also biochemical examination for serum total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, blood glucose and creatinine. HbA1c was determined by Bio-STAT. RESULTS: The rates of normal urinary albumin, micro- and macroalbuminuria were 66.67%, 22.22% and 11.11% in these patients, respectively. In patients with a history of diabetes for more than 10 years, the rates were 58.33%, 8.33% and 33.33%, respectively. CONCLUSION: Nearly 1/3 of type 2 diabetic patients with hypertension have microalbuminuria and proteinuria, and screening for microalbuminuria and proper interventions in these patients might prove beneficial.