Objective To compare three methods for the extraction of mycobacterial DNA.Methods Two commercial DNA extraction kits and an ordinary freeze-thawing method were used to extract DNA from the pure suspensions of three species of Mycobacteria (M.tuberculosis,M.leprae and M.smegmatis) at different densities (1 × 10 to 1 × 105 cells/ml),simulated clinical specimens containing different concentrations of mycobacterial cells (1 × 10 to 1 × 104 cells/ml).The purity and concentration of the extracted DNA were evaluated.Then,PCR was performed to amplify the 16S rRNA region of Mycobacteria.The performance of the three methods was compared by the purity and concentration of extracted DNA as well as the results of PCR.Further more,76 clinical skin specimens suspected to be infected with Mycobacteria were used to further validate the performance of these methods.Results All the extracted DNA samples could be detected by PCR.The highest purity of DNA was obtained by the kit A,followed sequentially by the freeze-thawing method and the kit B.When pure suspensions were used,the detection limit was consistently 1 × 102 cells/ml for all the three methods.With simulated specimens,the detection rate was consistently 100％ for all the three methods at the concentration of 1 × 103 cells/ml,60％ (12/20),55％ (11/20) and 55％ (11/20) for the kit A,kit B and freeze-thawing method respectively at the concentration of 1 × 102 cells/ml.The analysis of clinical specimens showed that the kit B could be used to extract DNA from paraffin-embedded specimens,with the detection rate similar to that of kit A and freeze-thawing method.Conclusions The kit A could rapidly yield high-quality genomic DNA of Mycobacteria by repeated cleaning of columns,and may serve as the optimal method for scientific and clinical studies,and the kit B is suitable for extracting mycobacterial DNA from fresh tissue specimens besides paraffin-embedded specimens.

OBJECTIVETo increase the reproduction efficiency of Iris plants.

METHODPollen viability, stigmatic receptivity, the color of anther and stigma of 5 Iris plants were observed during blooming.

RESULT1. The highest pollen viability was in 4 hours after blooming; 2. The stigmatic receptivities of I. sichuanensis, I. leptophylla, I. lactea and I. goniocarpa were strong in 4 hours after blooming, while that of I. lactea var. chinensis was strong in 2 hours after blooming; 3. The color of anther could reflect the pollen viability, but could not indicate the viability level; 4. The stigma color could not reflect the receptivity of stigma.

CONCLUSIONThe optimum artificial pollination time of these five species were 12:00 -14:00.

Biological Evolution ; Color ; Flowers ; physiology ; Iris Plant ; physiology ; Odorants ; Plant Infertility ; Pollen ; growth & development ; Pollination ; physiology ; Reproduction ; physiology ; Species Specificity

OBJECTIVETo identify risk factors for gestational diabetes mellitus (GDM) in women and to study the contribution of family history of type-2 diabetes to the risk for DDM.

METHODSA case-control study was performed in 85 women with GDM and 177 cases controls. Univariate and multivariate logistic regression and log-linear model were used to identify risk factors of GDM.

RESULTSMultivariate logistic regression showed that obesity before pregnancy, family history of type-2 diabetes, birth weight of pregnant women, age, fasting plasma level of triglyceride, physical inactivity, etc. all were risk factors for GDM. Analysis with log-linear model showed that parents' (father's or mother's) history of type-2 diabetes associated with GDM, with P-values of 0.012 and 0.017, respectively. Prevalence of diabetes in the mothers of proband with GDM was 9.41%, as compared with that in the fathers of proband with GDM 8.24%, with no statistical significance.

CONCLUSIONSObesity before gestation, family history of type-2 diabetes, low birth weight of mother, age, increased fasting plasma level of triglyceride, as well as parents' history of type-2 diabetes, all were risk factors for GDM. Physical exercise was found to be a protective factor for GDM. Mother's history of type-2 diabetes did not differ from father's in contributing to the onset of GDM in their offspring.

Adult ; Birth Weight ; Diabetes Mellitus ; genetics ; Diabetes, Gestational ; etiology ; Exercise ; Female ; Humans ; Logistic Models ; Obesity ; complications ; Pregnancy ; Risk Factors ; Triglycerides ; blood

OBJECTIVE: To investigate effect of hyperbaric oxygen (HBO) therapy on treatment of late healed wounds after pharyngeal and laryngeal carcinomas surgery. METHOD: Eighty-three patients with late healed wounds after pharyngolaryngectomy were divided into two groups, which were HBO treatment group (n = 48) and control group (n = 35) by random. 48 cases were treated with the general therapeutic measure along with HBO (the pressure is 2 x 10(5) Pa, 110 minutes per day for 5 to 10 days). The other 35 cases were treated with the general therapy without HBO as control. RESULT: There were fresh granulation erupted from the wound of entry after 3 or 4 days of HBO) treatment, then the granulation and the new blood capillary erupted continually to fill the wound surface, and the wound of entry was healed over finally. In the HBO and the control groups, the average healing time was 27.50 and 45.00 days (P < 0.01) in the wound by necrosectomy of the myocutaneous flap or forearm flap, 8. 50 and 14.09 days (P < 0.01) in the pharyngeal fistula, 5.93 and 8.62 days (P < 0.01) in the infected or fluid-filled wound. CONCLUSION HBO therapy is an effective and powerful treatment for late healed postoperative wounds in pharyngeal and laryngeal carcinomas surgery.
Adult ; Aged ; Female ; Humans ; Hyperbaric Oxygenation ; Laryngeal Neoplasms ; surgery ; therapy ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; Postoperative Period ; Treatment Outcome ; Wound Healing

Objective:To explore the effect and mechanism of lncRNA SBF2-AS1 on glioma cell proliferation, apoptosis and radiosensitivity.Methods:Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression levels of SBF2-AS1, miR-1287-5p and FSCN1 in normal human brain glial cell HEB and glioma cell lines, including LN18, SW1088, Hs683, and western blot method was used to detect the expression level of FSCN1 protein in cells. Glioma cells including LN18 and SW1088 were taken as the research object. After SBF2-AS1 small interfering RNA, miR-1287-5p mimics, FSCN1 small interfering RNA were transfected into LN18 cells, CCK-8 was used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, clone formation experiment was used to detect cell radiosensitivity, Western blot was used to detect the protein expressions of cyclin D1, cleaved-caspase 3 and phosphorylated histone 2A variant phosphorylated histone (γ-H2AX). Dual luciferase reporter gene experiments verified the regulatory relationship between SBF2-AS1 and miR-1287-5p, as well as miR-1287-5p and FSCN1.Results:Compared with HEB cells, the expression level of SBF2-AS1 and the expression levels of FSCN1 mRNA and protein in glioma cell line LN18, SW1088 and Hs683 were significantly increased ( P<0.05), and the expression level of miR-1287-5p was significantly reduced ( P<0.05). After down-regulating SBF2-AS1, up-regulating miR-1287-5p or down-regulating FSCN1 expression, LN18 and SW1088 cells activity and cyclin D1 protein expression were significantly reduced ( P<0.05), the apoptosis rate and cleared-caspase-3 protein expression were significantly increased ( P<0.05), the survival score was significantly reduced ( P<0.05), and the expression ofγ-H2AX protein was significantly increased ( P<0.05). The results of dual luciferase reporter gene assay showed that the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-WT or FSCN1-WT was lower than that of co-transfected miR-NC and SBF2-AS1-WT or FSCN1-WT in LN18 and SW1088 cells ( P<0.001), while the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-MUT or FSCN1-MUT was not significantly different from that of miR-NC transfected with SBF2-AS1-MUT or FSCN1-MUT ( P>0.05). The expression level of miR-1287-5p in LN18 and SW1088 cells in si-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), and the expression level of miR-1287-5p in LN18 and SW1088 cells in pcDNA-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), but lower than that of pcDNA-NC group ( P<0.05). The protein expression level of FSCN1 in LN18 and SW1088 cells in the miR-1287-5p group was significantly lower than that in the miR-NC group ( P<0.05), and the protein expression level of FSCN1 in LN18 and SW1088 cells in the anti-miR-1287-5p group was significantly higher than that in the anti-miR-NC group ( P<0.05). When miR-1287-5p and SBF2-AS1 were down-regulated simultaneously or FSCN1 was up-regulated while SBF2-AS1 was down-regulated simultaneously, the proliferation and cyclin D1 protein expression of LN18 and SW1088cells were significantly increased ( P<0.001), the apoptosis rate and cleared-caspase-3 protein expression were significantly reduced ( P<0.001), the survival score was significantly increased ( P<0.001), and the expression of γ-H2AX protein was significantly reduced ( P<0.001). Conclusion:SBF2-AS1 highly expresses in glioma cells, down-regulation of SBF2-AS1 expression can inhibit the proliferation of glioma cells, promote apoptosis, and enhance cell radiosensitivity by regulating the miR-1287-5p/FSCN1 axis.

Objective:To explore the effect and mechanism of lncRNA SBF2-AS1 on glioma cell proliferation, apoptosis and radiosensitivity.Methods:Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression levels of SBF2-AS1, miR-1287-5p and FSCN1 in normal human brain glial cell HEB and glioma cell lines, including LN18, SW1088, Hs683, and western blot method was used to detect the expression level of FSCN1 protein in cells. Glioma cells including LN18 and SW1088 were taken as the research object. After SBF2-AS1 small interfering RNA, miR-1287-5p mimics, FSCN1 small interfering RNA were transfected into LN18 cells, CCK-8 was used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, clone formation experiment was used to detect cell radiosensitivity, Western blot was used to detect the protein expressions of cyclin D1, cleaved-caspase 3 and phosphorylated histone 2A variant phosphorylated histone (γ-H2AX). Dual luciferase reporter gene experiments verified the regulatory relationship between SBF2-AS1 and miR-1287-5p, as well as miR-1287-5p and FSCN1.Results:Compared with HEB cells, the expression level of SBF2-AS1 and the expression levels of FSCN1 mRNA and protein in glioma cell line LN18, SW1088 and Hs683 were significantly increased ( P<0.05), and the expression level of miR-1287-5p was significantly reduced ( P<0.05). After down-regulating SBF2-AS1, up-regulating miR-1287-5p or down-regulating FSCN1 expression, LN18 and SW1088 cells activity and cyclin D1 protein expression were significantly reduced ( P<0.05), the apoptosis rate and cleared-caspase-3 protein expression were significantly increased ( P<0.05), the survival score was significantly reduced ( P<0.05), and the expression ofγ-H2AX protein was significantly increased ( P<0.05). The results of dual luciferase reporter gene assay showed that the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-WT or FSCN1-WT was lower than that of co-transfected miR-NC and SBF2-AS1-WT or FSCN1-WT in LN18 and SW1088 cells ( P<0.001), while the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-MUT or FSCN1-MUT was not significantly different from that of miR-NC transfected with SBF2-AS1-MUT or FSCN1-MUT ( P>0.05). The expression level of miR-1287-5p in LN18 and SW1088 cells in si-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), and the expression level of miR-1287-5p in LN18 and SW1088 cells in pcDNA-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), but lower than that of pcDNA-NC group ( P<0.05). The protein expression level of FSCN1 in LN18 and SW1088 cells in the miR-1287-5p group was significantly lower than that in the miR-NC group ( P<0.05), and the protein expression level of FSCN1 in LN18 and SW1088 cells in the anti-miR-1287-5p group was significantly higher than that in the anti-miR-NC group ( P<0.05). When miR-1287-5p and SBF2-AS1 were down-regulated simultaneously or FSCN1 was up-regulated while SBF2-AS1 was down-regulated simultaneously, the proliferation and cyclin D1 protein expression of LN18 and SW1088cells were significantly increased ( P<0.001), the apoptosis rate and cleared-caspase-3 protein expression were significantly reduced ( P<0.001), the survival score was significantly increased ( P<0.001), and the expression of γ-H2AX protein was significantly reduced ( P<0.001). Conclusion:SBF2-AS1 highly expresses in glioma cells, down-regulation of SBF2-AS1 expression can inhibit the proliferation of glioma cells, promote apoptosis, and enhance cell radiosensitivity by regulating the miR-1287-5p/FSCN1 axis.

Objective To explore the feasibility of the digital grading scale applied for assessment of anxiety by nurses. Methods Firstly, the digital grading scale was made, and then it was applied for patients guided by nurses to evaluate their anxiety level by themselves within 24 hours and at 3 weeks after admission. Anxiety level of the same crowd guided by doctors was also evaluated independently with Hamilton Anxiety Scale ( HAMA) by blind method. Results The correlation coefficent of the score between the digital grading scale and HAMA was 0. 794 (P<0. 01), and the score between them was positively related. Scores of the digital grading scale were compared with HAMA whose scores were divided into 5 groups according to severity level with significant differences (P <0. 01). Conclusions The digital grading scale has an obvious correlation with HAMA and it can be used to evaluate anxiety level in patients with generalized anxiety disorder.

BACKGROUND/AIMS: Early adverse life events (EALs) are relevant to irritable bowel syndrome in adulthood. Maternal separation (MS), as one of the EALs, has proved to induce visceral hypersensitivity in adult rats. However, the effect of MS on visceral hypersensitvity from the post-weaning period to adulthood remains unknown. METHODS: One hundred and ten neonatal Sprague-Dawley rats were randomly divided into 2 groups: rats in the MS group were exposed to 3 hours daily MS on postnatal day (PND) 2–14; the normal control (NC) group remained undisturbed. Visceral sensitivity was determined by measuring the visceromotor response to colorectal distention on PND21, 35, and 56. Anxiety-like behaviors were measured by the open field test. RESULTS: Compared with NC rats, MS rats showed significant visceral hypersensitivity from the post-weaning period to adult. The proportion of visceral hypersensitive rats decreased with age from 87.5% to 70.0% in the female MS group and from 90.0% to 66.7% in the male MS group. The relative VMR ratio of MS and NC on PND21 was higher than PND35 and PND56. MS rats showed decreased ability of movement and exploration to the novel environment in the post-weaning period, obesity in the prepubertal period, and more anxiety-like behaviors in adulthood. CONCLUSIONS: MS can significantly affect visceral sensitivity and behaviors of rats in different age stages, especially in the post-weaning period. Visceral hypersensitivity of MS rats is more pronounced in the post-weaning period and slightly restored in adults. Thus, visceral hypersensitivity in the post-weaning period might play a more meaningful pathophysiologic role in the formation of adult irritable bowel syndrome.
Adult ; Animals ; Female ; Humans ; Hypersensitivity* ; Irritable Bowel Syndrome ; Male ; Maternal Deprivation ; Obesity ; Rats ; Rats, Sprague-Dawley

Objective To observe the clinical effect of combining extracorporeal shock waves with pulsed radiofrequency irradiation of the dorsal root ganglia in the treatment of spinal cord-derived abdominal pain. Methods A total of 88 patients were randomly divided into a control group, a shock wave group, a pulsed irradiation group and a combination group, each of 22. All of the patients were given etocoxib and pregabalin medication for 3 weeks. In ad-dition to the drugs, the shock wave group received extracorporeal shock wave therapy, and the irradiation group re-ceived pulsed radio frequency irradiation of the dorsal root ganglia. The combination group received both. A visual an-alogue scale was used to assess perceived pain. The quality of life short form 36 ( QOL-SF36) , Hamilton anxiety scale and Hamilton depression scale were administered to all 4 groups before the treatments and after 1, 4 and 12 weeks of the treatments. The development of diseases, gastrointestinal function, medical treatment and medical expenses of the 4 groups were observed for two years after the treatments. Results After 4 weeks of treatment, the average scores of all four groups in all of the evaluations had improved significantly compared with before the treatment. In combination group′s average results were significantly better than those of the other 3 groups from 4 weeks until 12 weeks after the treatment. During the subsequent two years that group′s gastrointestinal symptoms, hospital visits and medical expen-ses were all significantly lower, on average, than those of the other groups. Conclusion Extracorporeal shock waves combined with pulsed radio frequency irradiation of the dorsal root ganglia has significant clinical efficacy for alleviating spinal cord-derived abdominal pain. It can significantly reduce medical costs and is worthy of clinical pro-motion.

Objective To analyze the epidemiological characteristics of population-based cohort of patients with the comorbidity of cardiovascular and cerebrovascular diseases and type 2 diabetes in Lingnan area,and to study the related influencing factors in the onset and progression of the disease. Methods A retrospective cohort study was used to collect data from people who underwent physical examination in the Eleventh People's Hospital of Guangzhou from May 2022 to December 2023. Data mainly included questionnaire surveys,physical examinations,and laboratory testing indicators. The 2022 was defined as the baseline to statistically analyze the occurrence and development of the comorbidity of cardiovascular and cerebrovascular diseases and type 2 diabetes in this population,and to analyze the related influencing factors of comorbidity and distribution of traditional Chinese medicine constitution in comorbidity population. Results Finally,a total of 26498 subjects were included,from which there were 359 patients with the comorbidity of cardiovascular and cerebrovascular diseases and type 2 diabetes (comorbidity group),accounting for 1.4％ of the total. Among them,290 were male,accounting for 80.8％,which is much higher than female. The mean age was(61.6±9.5)years old,which was significantly higher than that of the non-comorbidity group. The cases of comorbidity group were mainly concentrated in the age group of 45-75 years old,and no cases were found in people under 35 years old. There were 293 patients with the comorbidity of ischemic cardiovascular disease and type 2 diabetes,whose proportion (81.6％) is much higher than that of other types. Significant differences between comorbidity group and non-comorbidity group were found in terms of gender,age,age distribution,height,body mass,body mass index (BMI),smoking,alcohol consumption,marital status,exercise,and dampness syndrome (P<0.05). About 1.0％ of population at the baselined converted from non-comorbidities or single disease to comorbidities. The proportion of newly diagnosed patients with the comorbidity of ischemic cardiovascular disease and type 2 diabetes is the highest,up to 68.9％. BMI overweight or obesity,large waist circumference,smoking,dampness syndrome and exercise were the risk factors affecting the comorbidity of cardiovascular and cerebrovascular diseases and type 2 diabetes. A total of 264 cases of comorbidity group had finished evaluation of traditional Chinese medicine body constitutions. The proportion of balanced constitution was the highest (31.1％),followed by dampness-heat constitution (18.2％),yang-deficiency constitution (13.3％) and phlegm-dampness constitution (11.7％). Conclusion The incidence of the comorbidity of cardiovascular and cerebrovascular diseases and type 2 diabetes is high in Lingnan area,which may be related to dampness constitution,BMI overweight or obesity,large waist circumference,smoking,dampness syndrome and lack of exercise.