OBJECTIVE To investigate the resistant mechanisms of Candida albicans to azoles at molecular level.METHODS NCCLS M-27 protocols were used to test the in vitro susceptibilities of 102 C.albicans strains isolated from the patients with recurrent vulvovaginal candidiasis(RVVC) against fluconazole(FLC) and itraconazole(ITC) to screen the FLC-and ITC-resistant C.albicans isolates;six pairs of primers,A1-A2,B1-B2,C1-C2,D1-D2,E1-E2 and F1-F2 were respectively to amplify gene CYP51 of 4 strains with FLC-and ITC-resistance.The PCR products were sequenced and analyzed to identify the mutation sites by compared with the sequence of gene CYP51 of referenced C.albicans strain in NCBI site of Internet.RESULTS The analysis of full length sequence of CYP51 from 4 FLC-and ITC-resistant strains showed that from total 32 mutation sites there were 4 significant site mutations,where the mutation of GAT to GAC at 116 caused the substitution of D by E(E266D in two strains);GCC to GGT at 117 caused the substitution of A by G(A117G in 1 strain);GAA to GAC at 266 caused the substitution of E by D(E266D in 2 strains);and GTT to ATT at 488 caused the substitution of I by V(V488I in 1 strain).The site mutations of 266 and 488 were tested in 1 strain of 4 strains.CONCLUSIONS The CYP51 total gene of 4 strains has been checked out.Of FLC and ITC-resistant C.albicans alignment in this time,find out 4 significant bp mutations.Causing its amino acide change,among them,A117G has not be interrelated report still now.The details of mechanism need to be further studied.

Objective:To explore the application of the pioneering TEBMCL (task based evidence-based laboratory medicine team cooperative learning) teaching mode in the teaching of Clinical Biochemical Laboratory Technology. Methods:A total of 336 undergraduates of medical laboratory from 3 classes of Batch 2014 were selected as research objects, and 112 students in class 1 were randomly selected as the teaching reform class, 113 students in class 2 were randomized into control class 1 and 111 students in class 3 into control class 2. The teaching effects achieved by applying the TEBMCL teaching mode to the teaching of Clinical Biochemical Laboratory Technology in the three classes were compared comprehensively and the TEBMCL teaching mode was also evaluated. The Access 2010 and SPSS 17.0 statistical analysis methods were used for statistical analysis. Results:Learning effect: ①Compared with the basic test of the three classes, there was no statistical differences ( P > 0.05); but the in-class test after each chapter showed that the results of the teaching reform class were better than those of the other two classes ( P < 0.05). It was also found that the performance of the control class 1 was better than that of the control class 2 after applying the new TEBMCL teaching mode in the second chapter ( P < 0.05). ②In a number of indicators, it was found that the scores of the reform class were better than those of the control class 1 and the control class 2 ( P < 0.05); ③In the terms of excellent grade and good grade, the teaching reform class was ahead of the control class 1 and the control class 2, and none of students in the teaching reform class failed the graduation design (thesis). In the terms of questionnaires and seminars: TEBMCL teaching mode was carried out for juniors who had professional basic knowledge, with short prcoess and high efficiency, while this method still needed to be improved. Conclusion:The new TEBMCL teaching mode has significant advantages compared with the traditional teaching mode, which can better promote the cultivation of students' abilities and improve the degree of knowledge mastery.

Objective To study the effect of early entreat nutrition (EN) standardized treatment on optimization of blood glucose control and prognosis in acute respiratory distress syndrome (ARDS) patients with mechanical ventilation (MV). Methods Forty-two patients with MV of ARDS admitted to Huzhou First Municipal People's Hospital from April 2015 to March 2017 were enrolled. April 1st, 2016 was taken as the time node, the patients treated from April 1st, 2015 to March 31st, 2016 were assigned in the control group (n = 20), while the patients treated from April 1st, 2016 to March 31st, 2017 were included in the experimental group (n = 22). The patients in experimental group were given conventional treatment, in 24-48 hours after admission gastrointestinal decompression was stopped and early EN was begun through a nasointestinal tube; the patients in control group received conventional treatment and routine EN (given 48 hours after admission). The differences in nutritional support indexes, the blood glucose variability indexes and the prognostic related indicators were compared between the two groups. Results Compared with the control group, the initiation time for EN tolerance, first defecation time, time of reaching target feeding amount were significantly earlier in the early EN standardized treatment process management [time of initial EN tolerance (hours): 106.82±42.84 vs. 157.29±56.76, first defecation time (hours): 71.29±23.43 vs. 104.69±26.94, time of reaching target feeding amount (days): 6.24±1.25 vs. 9.86±2.36], the proportions of EN/EN+parenteral nutrition (PN) and the nasointestinal tube feeding reaching the standard on 7 days in experimental group were significantly increased [the proportion of EN/EN+PN:98.69% vs. 78.69%, the nasointestinal tube feeding reaching standard: 68.18% (15/22) vs. 45.00% (9/20)], average level of blood glucose (GLUave), maximum value of blood glucose (GLUmax), standard deviation of blood glucose (GLUsd), coefficient of variation of blood glucose (GLUcv), hyperglycemia incidence, incidence of multiple organ dysfunction syndrome (MODS), 28-day mortality were significantly decreased [GLUave (mmol/L): 9.4±2.6 vs. 11.5±3.9, GLUmax (mmol/L): 14.19±2.36 vs. 16.26±4.89, GLUsd (mmol/L): 4.86±1.27 vs. 6.87±2.46, GLUcv: (49.86±6.32)% vs. (59.95±5.81)%, hyperglycemia incidence: 59.09% (13/22) vs. 80.00% (16/20), incidence of MODS: 59.09% (13/22) vs. 80.00% (16/20), 28-day mortality: 36.36% (8/22) vs. 45.00% (9/20)], minimum value of blood glucose (GLUmin) was significantly increased (mmol/L: 5.86±2.32 vs. 4.18±1.86), invasive MV time was significantly shorted (hours:156.82±26.84 vs. 169.93±32.34) with statistically significant differences (all P < 0.05). Early EN could also improve the patient's pulmonary oxygenation function. Since 9 days of disease course, the oxygenation index (PaO2/FiO2) in the experimental group was significantly higher than that of the control group [mmHg (1 mmHg = 0.133 kPa): 256.97±18.63 vs. 239.82±21.72, P = 0.068], but there was no significant difference in the length of ICU stay (days: 13.9±3.6 vs. 14.8±3.4, P > 0.05). Conclusion The early EN standardized treatment process management can improve the nutritional status, decrease blood sugar fluctuations, and further benefit the improvement of the prognosis of ARDS patients with MV.

Objective:To investigate the clinical characteristics and treatment of patients with refractory spasm tetanus.Methods:Tetanus patients admitted to the Affiliated Hospital of Zunyi Medical University from January 2011 to April 2021 were collected and divided into the refractory spasm group and general group according to whether they were refractory spasm. The general demographic characteristics, clinical characteristics, treatment and prognosis of the two groups were compared. The treatment and risk predictors of patients with refractory spasm tetanus were explored.Results:Among the 59 tetanus patients, 35 patients (59.32%) were in the refractory spasm group and 24 patients (40.68%) were in the general group. There were no significant differences in sex, age, latency and trauma site between the two groups ( P>0.05). All patients with refractory spasmodic tetanus were treated with mechanical ventilation, the length of hospital stay was significantly prolonged, and the rate of pulmonary infection was significantly increased ( P<0.05). The incidence of multiple sites muscular rigidity (spasm/limb stiffness, neck stiffness, and angular pedicle tension) in patients with refractory spasmodic on the first day of admission was higher than that in the general group ( P<0.05). The patients' initial symptoms (within 24 h after admission) were muscle spasm/limb stiffness + neck stiffness + angular pedicle inversion, which had the highest specificity for predicting the occurrence of refractory spasm. The proportion of Ablett grade Ⅳ in patients with refractory spasm was higher than that in the general group, and the difference was statistically significant ( P<0.05). The 48.75% patients with refractory spasm tetanus were treated with more than 3 sedatives combined with muscle relaxants, and the duration of sedative use was significantly prolonged ( P<0.05). Conclusions:The mechanical ventilation time and hospitalization time in tetanus patients with refractory spasm are significantly prolonged, and the incidence of pulmonary infection is significantly increased, which requires the combined application of a large number of sedative and muscle relaxants for a long time, and the incidence of refractory spasm is higher in patients with multi-site muscular rigidity at the early stage of the disease.

Objective: Alport syndrome was an inherited kidney disease caused by the mutation of COL4A3, COL4A4, or COL4A5. Whole-exome sequencing was used to detect the mutations on these genes for the molecular diagnosis of Alport syndrome. Methods: A 6-year-old girl found accidentally with microscopic hematuria at the age of 4. The clinical data and blood sample of the family including proband, parents, brothers, and sisters were collected. Whole exome sequencing was conducted using their genomic DNAs. Results: A novel heterozygous frameshift mutation c.1826delC (p.Pro609Glnfs*44) was found in the exon 25 of the COL4A4 (NM_000092) in the proband, the father, and the sister, showing an autosomal dominant inheritance pattern of Alport syndrome. This mutation of COL4A4 was confirmed by mutation analysis, and the mutation of c.1826delC was verified by Sanger sequencing. No mutations on COL4A3 and COL4A5 were detected in this family. And the mother and brother are normal wide-type. Conclusions This novel mutation is a valuable addition to the current genetic profile of Alport syndrome, and provide us a better understanding of the disease. Whole-exome sequencing is a power tool to identify the novel mutations of inherited disease and contribute to the molecular diagnosis of disease.