Objective To assess the application value of REP-PCR in genotyping of candida glabrata strains in clinical pratice. MethodsFrom 2009 to 2010, thirty-eight candida glabrata strains were isolated from Shanghai Ruijin Hospitals, Shanghai Renji Hospital, Shanghai Huashan Hospital, Anhui Medical University Hospital, Shenzhen People's Hospital. Six loci in housekeeping genes (FKS, LEU2,NMT1, TRP1, UGP1 and URA3 ) were amplified and sequenced. The sequences were compared with the MIST database and allele profile and sequence type (ST) were obtained. With primers Ca21, Ca22 and Com21 used to amplify the adjacent variable gene regions,the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Finally, the results of these two genotyping methods were compared. ResultsFor REP-PCR, Ca22-Com21 has the best genotyping effect. REP-PCR and MLST have the same genotyping results. Five REP-PCR types were found in 38 candida glabrsta isolates. Type A,B, C, D and E strains from REP-PCR were genotyped as ST 7, 3, 19, 45 and new type respectively byMIST. REP-PCR saves time compared with MIST. Conclusions REP-PCR offers a simple and rapid method for molecular typing, with similar discriminatory power with MIST. Therefore, REP-PCR can be the preferred choice in laboratory, especially for a large number of isolates.

BACKGROUND:The blood supply is essential for the repair of the meniscus after the suture of it in the injured knee joint. According to the condition of blood supply, the meniscus was divided into three zones by Arnoczk in 1982. The absolute vascularization zone, in which the distance to the combination part of meniscus and synovial membrane is inferior to 3mm, is called the red zone. The absolute devascularization zone, in which the above-mentioned distance is superior to 5 mm, is called the white zone.And the relative vascularization zone with the distance between 3 mm to 5mm is called red-white zone.OBJECTIVE: To observe and discuss the anatomical features of the blood supplyof the meniscus.DESIGN:Single sample observation.SETTING:Department of Orthopaedics, Huaxi Hospital of Sichuan University.PARTICIPANTS: The study was conducted in Huaxi Hospital of Sichuan University during the period from September 2000 to March 2001.Nineteen cases were collected, including 19 knees (38 menisci) from the subjects, 17 male and 2 female, whose age varied from 21 to 43 years old.All the samples were obtained in the amputation operation, the isolated limbs were without the change of re-transplantation and the voluntary donations from the non-minorities living in Sichuan.meniscus samples: 7 knees (14 menisci) were taken. After the femoral artery intubation and infusion of the heparinized normal saline, the vascular bed had no blood and then was infused with the normal saline and ink of 0.5 volume fraction and the infusion pressure was 13 kPa. The infused meniscus samples underwent the dehydration of gradient ethanol,the transparency of dimethylbenzene, and preservation with ilex pedunculosa, and then the blood supply of the menisci was observed and distribution of the meniscus section: 6 knees (12 menisci) with fresh and uninfused menisci were taken. The sections obtained at the sagittal plane,the coronal plane and the horizontal plane underwent hematoxylin-eosin staining. Then the tissues structure and blood supply of the meniscus samples were observed with light microscope. Another 6 knees (12meniscus) underwent the specific CD34 antibody immunohistochemical staining. Then the tissue structure and blood supply distribution of the meniscus samples were observed with the light microscope.in the devascularization zone of the meniscus.RESULTS: Nineteen cases with 38 menisci all went into the final result resourced from the medial and lateral genicular arteries and the middle genicular artery. The small vessels from them form the peripheral capillary vessel plexus and annular vessel meshwork in the synovium and joint capsule and then go into the menisci for the nutrition of the lateral The apparent lunar artery is sent by the vessel plexus around the meniscus into the body of the meniscus and arranges in the inferior,middle and superior layers and dominates 25%-30% of the zone around The vessels of the angular part of the semilunar plate cover the whole meniscus. The nutrition is supplied by the membranous synovium on the surface of the meniscus.Rich blood supply was observed in the anterior and posterior angular laminated blood supply in the body of the meniscus was newly devascularization zone of the meniscus is supplied from the joint synovial fluid is explicit.

OBJECTIVE:To prepare Zuojin gastric-mucoadhesive tablets and evaluate their drug release properties in vitro. METHODS:Zuojin gastric-mucoadhesive tablets were prepared with hydroxypropyl methyl cellulose K15M(HPMC-K15M),car-bomer 934P and HPMC-E50 as the bioadhesive and matrix materials,and basic magnesium carbonate(foaming material),95% al-cohol solution(adhesive)and aerosil(glidant and lubricant)as the adjuvants. With the accumulative release of total alkaloids from Coptis chinensis Franch. at 2,6 and 10 h(Q2 h,Q6 h and Q10 h)as the indexes,orthogonal design test was conducted to optimize the amounts of HPMC-K15M,carbomer 934P,HPMC-E50 and basic magnesium carbonate,and verification was carried out. Drug re-lease properties in vitro of the preparation and Zuojin conventional tablets were observed and in vitro adhesion thereof determined. RESULTS:The optimal formulation was as follows as that for 50 tablets,HPMC-K15M of 0.7 g,carbomer 934P of 0.2 g, HPMC-E50 of 3.5 g and basic magnesium carbonate of 0.4 g. The Q2 h,Q6 h and Q10 h of three batches of prepared samples were 24.32%,56.10% and 77.04% respectively. 1-12 h drug release in vitro of prepared samples was in conformity with Ritger-Peppas equation. The Q2 h of Zuojin conventional tablets and Zuojin gastric-mucoadhesive tablets were 80.46% and 24.04%,Q12 h thereof 92.15% and 95.83% and gastric adhesion thereof 24.2 and 74.0 g/cm2,respectively. CONCLUSIONS:Zuojin gastric-mucoadhesive tablets which have sustained-release effect and adhesive property have been prepared successfully.

Objective To investigate the role of PDR1 gene in azole-resistant Candida glabrata (C.glabrata).Methods Thirty-eight clinical isolates of C.glabrata were collected from five different hospitals.The minimal inhibitory concentrations (MIC) of azole antifungals including fluconazole,itraconazole and voriconazole against C.glabrata were determined by broth microdilution.Sequencing and amplification of PDR1 gene was achieved by real-time quantitative polymerase chain reaction (PCR).The mutation was cloned into an expression plasmid and then transferred into C.glabrata.The efflux of rhodamine 6G and drug sensitivity test were performed,and expressions of CDR1 and CDR2 were examined to verify function of mutation.Results Among these 38 isolates of C.glabrata,17 were resistant to at least one of azole antifungals.Moreover,mutations of PDR1 gene existed in every resistant isolates.Results of phenotyping test showed that in the isolate that expressed PDR1P927S,the expression of CDR1 and CDR2 were increased by 20.53 and 4.03 fold,respectively.And the fluorescence intensity of rhodamine 6G was decreased to 0.62 in efflux experiment.Conclusion P927S mutation of PDR1 gene could induce azole resistance of C.glabrata by increasing the expressions of CDR1 and CDR2,which results in drug resistance due to enhanced effect of efflux pump.

Objective To analyze the effect of bifrontal decompressive craniectomy on patients with refractory diffusing of brain swelling after severe traumatic brain injury.Methods The clinical data of 68 patients with refractory diffusing of brain swelling after severe traumatic brain injury were analyzed retrospectively.Thirty-five patients were performed with bifrontal decompressive craniectomy as observed group,continued intracranial pressure monitoring after surgery.Thirty-three patients were treated conservatively to reduce intracranial pressure as control group,continued intracranial pressure monitoring.The Glasgow outcome scale after discharge 6 months were assessed.The efficacy,the incidence of complications were observed in observed group.Results The admission intracranial pressure in observed group was significantly higher than that after surgery [(35.9 ±6.9) mmHg (1 mmHg =0.133 kPa) vs.(17.5 ±5.2) mmHg,P ＜0.05].The admission intracranial pressure in control group was (34.2 ± 8.6) mmHg,after admission 10.5 h was (32.0 ±4.8) mmHg (P ＜0.05),difference was no statistically significant (P＞ 0.05).The intracranial pressure after admission 10.5 h in control group was significantly higher than that in observed group after surgery (P ＜0.05).Two cases of subdural effusion,1 case of postoperative hydrocephalus in observed group.The better prognosis rate in observed group was significantly higher than that in control group [45.7％ (16/35) vs.18.2％ (6/33),P ＜ 0.05].Conclusions Bifrontal decompressive craniectomy is a suitable measure to decrease the intracranial pressure in the patients with refractory diffusing of brain swelling.If carried out early,it could provide better outcome for these patients.

BACKGROUND:Subcutaneous effusion often occurs after digital shaping titanium mesh cranioplasty, and affects therapeutic effects.
OBJECTIVE:To explore the causes and corresponding prevention measures of subcutaneous effusion after digital shaping titanium mesh cranioplasty.
METHODS: We retrospectively analyzed the clinical data and treatment methods of 19 cases of subcutaneous effusion after digital shaping titanium mesh cranioplasty, summarized the postoperative complications and explored the effective methods for prevention and treatment of subcutaneous effusion.
RESULTS AND CONCLUSION: After active treatment, five cases of dural breakage, four cases of foreign body stimulation, three cases of getting out of bed early, three cases of early extubation, three cases of long-time operation repair, and one case of excessive use of electric knife were al cured. Dural breakage and foreign body stimulation is considered as the main causes of postoperative effusion. Patients with subcutaneous effusion were given releasing elastic bandage, aspiration, and strict bed rest. After these active treatments, patients were al cured. Subcutaneous effusion may result from single or mixed factors. The above-mentioned causes are only a part. Non-central suspension, incomplete hemostasis, and preoperative excessive colapse of the bone window are al reported to be the reasons for the occurrence of subcutaneous effusion.

Objective To investigate the mechanisms of fluconazole resistance in clinical and experimental induced isolates of C.glabrata.Methods Efflux of rhodamine 6G was performed to evaluate the effects of efflux pumps.The expression levels of transporter genes CDR1,CDR2,SNQ2 and ERG11 were examined by real-time RT-PCR.Meanwhile,sequence of PDR1 was determined by PCR based DNA sequencing.Results Efflux pumps of all fluconazole-resistant isolates had stronger effects than that of susceptible isolates,consistently with significant upregulation of CDR1,but no obvious difference was found in CDR2 or SNQ2.Also,no notable change in the expression level of ERG11 between susceptible and resistant isolates.PDR1 mutations existed in both clinical and experimental induced isolates of C.glabrata,among which P927S,L543P and S947L haven't been reported previously.Conclusion Mutations of PDR1 were induced by fluconazole both in vivo andin vitro,which will result in overexpression of CDR1 and strengthen the effect of efflux pump.

Objective To evaluate the reliability and validity of the Chinese version of Uncertainty Scale for Kids-22 (USK-22).Methods The Uncertainty Scale for Kids was translated and adapted according to Chinese culture.The reliability and validity of the Chinese version of USK were tested in 117children and adolescents with cancer.Results The rate of recovery was 100％.The rote of effective questionnaire was 95.7％.The patients were easily to understand and it cost them average 8 minutes to finish the questionnaires.Three factors were extracted by factor analysis and could explain 57.043％ of total variance.The correlations between each item and subscale ranged from 0.526 to 0.762.The Cronbach α of USK was 0.872 and the Cronbach α of each subscale ranged from 0.774 to 0.826.The testretest reliability was 0.835.Conclusions The Chinese version of USK is reliable and valid,and can be used to measure the level of uncertainty among Chinese children and adolescents with cancer.

Objective The Study the difference of miRNA expression of AIDS patients with toxic heat accumulation syn -drome with healthy control group .Methods This research used Agilent miRNA chip to test the miRNA of blood preparation ,and used SAS system to screen the differences between groups ,then analyzed the significance function of target gene .Results Compare the heat-toxic accumulation group and the healthy control group ,100 cases of obvious expression difference and of which the differ-ence was 2 times and greater were screened out (in which 64 cases showed up-regulation and 36 showed down-regulation) .Differen-tially expressed miRNAs function involves the role of IL-21 receptor in T cell activation ,C reactive protein ,and the positive regula-tion of the immune response .Conclusion AIDS toxic heat accumulation syndrome patients and healthy control groups exist differ -ences in miRNA expression profiles ,the biological basis of syndrome maybe related to T cell activation and IL-21 receptor .

Objective: To construct an evaluation index system of oral health literacy for children's parents, so as to provide an index system for the evaluation of oral health literacy for children's parents in China. Methods: The evaluation index system of oral health literacy for children's parents was designed based on literature review and semi-structured interview. Experts from oral prevention and pediatric oral medicine were invited to participate in two-round Delphi consultations. The indicators were scored and screened according to the importance, and the weight determined using analytic hierarchy process. The effectiveness of the consultation was evaluated by positive coefficient, authority coefficient and coordination coefficient. Results: Twenty-four experts participated in the consultation, including 6 males and 18 females. There were 21 experts with a master degree, 3 experts with a doctor degree, and 20 experts with vice senior professional titles and above. The recovery rates of the two rounds of consultations were 96.00% and 100.00%, the authority coefficients were 0.866 and 0.917, the Kendall's coefficient of concordance were 0.120 and 0.156 (both P<0.05), and the coefficient of variation was 0.15-0.38 and 0.03-0.17, respectively. The final evaluation index system included 3 primary indicators, 11 secondary indicators and 40 tertiary indicators. The primary indicators were basic knowledge and concepts related to oral health, promoting lifestyle and behaviors related to oral health, and maintaining basic skills related to oral health. Conclusion The evaluation index system of oral health literacy for children's parents has been established in this study and needs to be further applied and evaluated.