To establish a simple, sensitive and effective technique for the identification of six common dermatophytes, polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene were used. The DNA of 6 common dermatophytes was amplified by primer dPsD1 and then primers dPsD2. The products generated by dPsD2 were digested with restriction enzyme Hinc II and Hinf I separately. A DNA fragment of about 3390 bp was amplified by using primer dPsD1 from the genomic DNA of each dermatophyte species. The product of dPsD2 was 2380 bp and the restriction profiles of Hinc II and Hinf I were between 58-1670 bp. By using PCR-RFLP, all of the 6 dermatophytoses were diagnosed to species level and no obvious difference identification between Hinc II and Hinf I. It is concluded that the PCR-RFLP identification of dermatophytes by Hinc II or Hinf I is efficient and rapid in clinical practice.
Arthrodermataceae/*classification ; Arthrodermataceae/genetics ; Arthrodermataceae/*isolation & purification ; DNA Topoisomerases, Type II/genetics ; DNA, Fungal/analysis ; DNA, Fungal/genetics ; Dermatomycoses/*microbiology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme Hinc II. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species-specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans. From the restriction profiles of Hinc II, 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCR-RFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase II gene is rapid and efficient.
Arthrodermataceae/classification ; Arthrodermataceae/*isolation & purification ; Aspergillus/*isolation & purification ; Candida albicans/isolation & purification ; DNA Topoisomerases, Type II/genetics ; Dermatomycoses/microbiology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Trichophyton/*isolation & purification

Objective To investigate the photo-toxic effects of indocyanine green (ICG) based on photodynamic therapy (PDT) on vascular endothelial cells in vitro. Methods Human umbilical vein endothelial cells were incubated with ICG with different concentrations in 96-well plate. According to the dose of ICG , they were divided into 6 groups as 0, 50, 100, 150, 200 and 250 μmol/L group. Each group consists of 8 wells, 4 of them were irradiated with a 810 nm laser of total power 24.0 J/cm2. Cell viability was measured by MTT assay 24 h after light irradiation. Morphological changes in cells were observed by light microscope. Results With the irradiation of 810 nm light , there was a significant decrease in relative cell viability in all groups except 0 μmol/L group (all P < 0). With the increase of concentration of ICG, the cell viability dropped rapidly (P < 0). Apoptotic changes can be seen in all groups except 0 μmol/L group and necrosis was the main change in cells of 200 μmol/L and 250 μmol/L group. Conclusion PDT with appropriate concentration of ICG has a strong inhibition to vascular endothelial cells in vitro and ICG-PDT may be a good new therapy for nevascularization.

Objective:To explore the relationship between angiotensin converting enzyme (ACE) gene insertion/dele‐tion (I/D) polymorphism and familial essential hypertension (EH) .Methods :Fluorescent in situ hybridization stai‐ning DNA sequencing test was used to detect ACE gene I/D genotype frequency and allele frequency in 46 familial EH patients (familial EH group) ,64 EH patients without family history (non‐familial EH group) and 43 healthy people (healthy control group) .Results:In healthy control group ,non -familial EH group ,familial EH group ACE gene DD genotype frequency was 11.6% ,32.3% and 37.0% ;distribution frequency of D allele was 33.7% , 52.3% and 57.6% respectively .Compared with healthy control group ,there were significant rise in ACE gene DD genotype and D allele frequency in familial EH group and non -familial EH group (P<0.05 or <0.01) ,but there were no significant difference between familial EH group and non‐familial EH group (P>0.05) .Conclusion:ACE gene DD genotype and D allele may be genetic predisposing genes of patients with essential hypertension ,and there is no significant difference in genetic constituent ratio between familial EH patients and non‐familial EH patients .

Paridis Rhizoma is a rare Chinese herbal medicine with variety of medicinal values. In recent years, the demand for Paridis Rhizoma has increased gradually. Artificial cultivation has met difficulties, while exploitation of wild resources was caught in a vicious circle and has overdrawn seriously. So it is of great significance to enhance the protection of Paridis Rhizoma, carry out basic research, in order to solve problems in seeding breeding, promote artificial cultivation to meet the market need and achieve sustainable development and supply. This article reviewed the status qua of seedling breeding of Paridis Rhizoma, including seed breeding, tuber breeding and tissue culture, with a purpose to standardize planting of resource conservation and utilization of Paridis Rhizoma.

The clinical data of patients with hypoparathyroidism accompanied by elevated and normal muscle enzymes were analyzed retrospectively.The results showed that there were no differences in age,gender,pathogenesis,etiology,and parathormone level between groups with elevated muscle enzyme and normal muscle enzyme.Serum calcium concentration in the group with normal muscle enzyme was within normal range.Serum calcium concentration in the group with elevated muscle enzyme was significantly lower than that in the group with normal muscle enzyme (P＜0.01).As the serum calcium concentration rose,the elevated enzyme level gradually returned to normal range,suggesting that the decrease of serum calcium concentration resulted in the elevated muscle enzyme levels in patients with hypoparathyroidism.

Objective To explore the effect of comprehensive management of cardiac rehabilitation on the quality of life of patients after percutaneous coronary intervention(PCI). Methods A total of 60 patients with PCI were randomly divided into the rehabilitation group and the control group with 30 cases each by convenience sampling method. The control group received routine treatment and nursing care of postoperative PCI, and the rehabilitation group received cardiac rehabilitation management on the basis of the control group, including training guidance, management of double heart psychological intervention, nutritional intervention diet guidance, evidence-based medicine, and set up system etc. The Hospital Anxiety and Depression Scale(HADS) score, 6 minutes walking distance (6MWD), coronary heart disease risk factors and Seattle Angina Score (SAQ score) were observed at 6 months after PCI. Results HADS score anxiety rate, depression rate, anxiety and depressive state at 6 months after PCI was 10.0%(3/30), 10.0%(3/30), 0 respectively in the rehabilitation group, and 33.3%(10/30), 36.7%(11/30), 13.3%(4/30) in the control group respectively, and there was significant difference between 2 groups (χ2=4.504, 5.963, 1.071. all P<0.05). 6MWD was (451 ± 48) m in the rehabilitation group and (402 ± 42) m in the control group, and there was significant difference between 2 groups (t=6.155, P=0.022). Compared two groups of coronary heart disease risk factors control level: there was significant difference in total cholesterol, triglycerides, high density lipoprotein cholesterol, low density lipoprotein cholesterol, glycosylated hemoglobin, body mass index, systolic pressure, diastolic pressure(t=-2.927-5.981, P<0.01 or 0.05). SAQ score in physical limitation, angina stability , angina pectoris, treatment satisfaction, disease awareness scores were (73.79 ± 6.04), (80.35 ± 7.63), (75.31 ± 8.11), (80.23 ± 5.44), (76.59 ± 14.29) points in the rehabilitation group, and (53.19 ± 5.13), (65.87 ± 18.85), (64.41 ± 6.31), (70.49 ± 8.01), (45.83 ± 8.65) points in the control group, there were significant differences between 2 groups (t=-21.630-18.959, all P<0.01). Conclusions The comprehensive management of cardiac rehabilitation alleviates the anxiety and depression of patients after PCI, reduces the risk factors of coronary heart disease, and improves the exercise ability and quality of life.

OBJECTIVETo explore the effect of Shengjiang Xiexin Decoction (SXD) on the intestinal mucosal and functional cells of rats after irinotecan (CPT-11) chemotherapy.

METHODSTotally 24 healthy Sprague-Dawley (SD) male rats were divided into three groups, the normal control group, the CPT-11 group, the SXD combined CPT-11 group according to random digit table, 8 in each group. CPT-11 was injected at the daily dose of 150 mg/kg to rats in the CPT-11 group and the SXD combined CPT-11 group from the caudal vein on the 4th day, once daily for 2 successive days to duplicate delayed diarrhea model. Equal volume of normal saline was injected to rats in the normal control group from the caudal vein. SXD at 2 g/mL (10 g/kg body weight) was administered to rats in the SXD combined CPT-11 group by gastrogavage for 9 successive days. Deionized water was administered to rats in the CPT-11 group and the normal control group. Diarrhea was observed at 48, 60, 72, 84, 96, and 108 h to calculate the incidence rate of diarrhea. Meanwhile, scoring for diarrhea was performed by referring methods of Akinobu Kurita. Rats were killed on day 10, ileum, cecum, and colon tissues were collected and fixed in 10% formalin solution. HE staining was performed. Intestinal mucosa injuries were graded under light microscope according to the criterion of Chiu's score. The expressions of goblet cells and Paneth cells were observed by PAS stain. Enteroendocrine cells were observed by immunohistochemical CgA staining. Positive cells were counted and cumulative optical density (IOD) analyzed by Image-Pro-Plus 6.0.

RESULTSNo diarrhea occurred in rats of the normal control group at each time point. The incidence rate of diarrhea was 75.0% (6/8) at 48 h, 100.0% (8/8) at 60 h, 100.0% (8/8) at 72 h, 87.5% (7/8) at 84 h, 75.0% (6/8) at 96 h, and 75.0% (6/8) at 108 h in the CPT-11 group. The incidence rate of diarrhea was 25.0% (2/8) at 48 h, 50.0% (4/8) at 60 h, 12.5% (1/8) at 72 h, 0.0% (0/8) at 84 h in the SXD combined CPT-11 group. Compared with the same group at 60 h, scores for diarrhea at 48, 84, 96, and 108 h obviously decreased in the CPT-11 group, and scores for diarrhea at 48, 72, 84, 96, and 108 h obviously decreased in the SXD combined CPT-11 group (P < 0.05, P < 0.01). Compared with the same group at 72 h, scores for diarrhea at 84, 96, and 108 h obviously decreased in the CPT-11 group (P < 0.05, P < 0.01). Compared with the normal control group, scores for diarrhea increased in the CPT-11 group at each time point (P < 0.01); grading of ileum, cecum, and colon mucosal tissues increased (P < 0.05, P < 0.01); expressions of ileum and cecum mucosal epithelial goblet cells obviously decreased (P < 0.05); the number and expressions of ileum and cecum mucosal epithelial Paneth cells increased (P < 0.01). Expressions of ilium endocrine cells increased, while those of cecum and colon endocrine cells decreased in the CPT-11 group (P < 0.01). Compared with the CPT-11 group, scores for diarrhea were obviously lowered (P < 0.05, P < 0.01), grading of ileum, and cecum mucosal tissues decreased (P < 0.05, P < 0.01); expressions of ileum, cecum, and colon mucosal epithelial goblet cells obviously increased (P < 0.05, P < 0.01); the number and expressions of ileum cecum mucosal epithelial Paneth cells increased (P < 0.05); expressions of cecum and colon endocrine cells increased (P < 0.05, P < 0.01) in the SXD combined CPT-11 group.

CONCLUSIONSXD played roles in preventing and treating CPT-11 induced delayed diarrhea by improving CPT-11 chemotherapy induced apoptosis and necrosis of intestinal mucosal and functional cells.

Animals ; Apoptosis ; Camptothecin ; adverse effects ; analogs & derivatives ; Colon ; Diarrhea ; Drug Therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Ileum ; Intestinal Mucosa ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Wound Healing ; drug effects

Objective To study the relationship between the methylation status of CPG islands in MEG3 gene promoter region of epithelial ovarian cancer and its clinical and pathological features. Methods The promoter methylation status was evaluated by MSP (methylation-specific polymerase chain reaction ) in 47 cases of ovarian cancer tissue and 15 cases of normal control. Results The methylation ratio (42.6%) of the MEG3 genes in the ovarian cancer was statistically significantly higher (P = 0.035 ) than that (13.3%) in the normal control. The methyation rate of the group with an age > 60 years old was slightly higher than that of the group with an age≤60 years old, without statistically significant (P > 0.05), so was observed in ovarian cancers of stage Ⅰ andⅡ than that in stage Ⅲ and Ⅳ. There were also no significant differences in MEG3 gene methylation positive rate neither in different pathological grading nor in various ovarian cancer tissues (P > 0.05). Conclusion Abnormal methylation in MEG3 gene may be associated with epithelial ovarian cancer , but no relation to its clinical pathology.

The interaction between medical instrument and target tissue during the surgery occurs in instrument-tissue interface. The reliability research on the interface is directly related to the safety and effectiveness of medical instrument in the clinical application. This paper illustrates the necessity of reliability research on instrument-tissue interface. Two main contents are synthetically reviewed the present paper: (1) reliability research on medical instruments; (2) biological tissue properties and its mechanical response.
Humans ; Reproducibility of Results ; Surgical Instruments ; Surgical Procedures, Operative