Objective To determine enantiomeric impurity of etomidate using high performance liquid chromatography. Methods (R)-etomidate and (S)-etomidate were isopropanol-n-hexane. The flow rate of the mobile phase was 0.5mL/min. The detected wavelength was 242nm. Results (R)-etomidate and (S)-etomidate could be separated completely under these conditions. The precision of (R)-etomidate was 1.57% (n=3). The limit of detection of (R)-etomidate was 4.25ng/mL. The average percentage content of (S)-etomidate was 0.09% in the samples. Conclusion The method was repeatable and sufficiently sensitive to determine the enantiomeric impurity of etomidate. It allows the quantitation of the impurities at the 0.085% (w/w) level relative to etomidate at a concentration of the test solution of 5mg/mL.

AIM: To explore a simple, reliable method for tissue processing and section staining by extracting DNA from the manually microdissectioned specimen, and to identify whether the extracted DNA is useful in the following study at molecule level. METHODS: The experiment was performed at the pathological laboratory of Guangdong Medical College from July 2004 to July 2007. The paraffin imbedding tissue sections of cervical cancer were thoroughly deparaffinized after mounted on slides for a long period of time. The nucleus was slightly stained with hematoxylin and microdissectioned under inverted microscope. The microdissectioned samples were put into EP tubes filled with digestion buffer to split the cells and then the DNA was extracted. During the whole course, PE tubes did not change, and the complicated phenol/chloroform extraction did not perform. The DNA extraction was rapid and simple. RESULTS: The DNA was measured by the spectrophotometer with concentrations from 0.14 to 5.25 g/L and absorbance values of A260/A280 were 1.6-1.8. All samples were amplified with PCR to produce expected length specific target fragment (231 bp). CONCLUSION: Rapid DNA extraction after manual tissue microdissection can produce adequate amount of DNA and maintain good quality of DNA template for PCR. The DNA meets the need of the following molecular experiments.

Objective To evaluate whether sofosbuvir (SOF)-based direct-acting antiviral agents (DAAs) combined with ribavirin (RBV) (combined RBV) can yield benefits to the recipients infected with hepatitis C virus (HCV) genotype 1 (GT1) after liver transplantation through systematic evaluation and Meta-analysis. Methods Multiple databases at home and abroad were systematically searched, the literature screening was conducted according to relevant standards, the quality of literatures was evaluated and data extraction was performed. The literature was divided into two groups according to the recipients with HCV-GT1 hepatitis after liver transplantation who received the treatment combined RBV or SOF-based DAAs alone without RBV (not combined RBV). Meta-analysis of the data was carried out using Rev Man 5.3 and R3.4.3 software. The incidence of sustained virological response 12 weeks (SVR12) after therapy was evaluated. Results A total of 2 195 articles were retrieved, and 6 articles published in English were eventually included according to the inclusion criteria. The Meta-analysis results demonstrated that the incidence of SVR12 did not significantly differ between the combined RBV and not combined RBV groups (P=0.28). However, the incidence of anemia in the combined RBV group was significantly higher than that in the other group (P < 0.01). Both combined RBV and not combined RBV therapies were efficacious in treating HCV-GT1a and HCV-GT1b subtypes after liver transplantation with similar clinical efficacy (P=0.33). The incidence of SVR in HCV-GT1 recipients did not significantly differ after receiving 12- and 24-weeks therapy after liver transplantation (P=0.95). Conclusions When SOF-based DAAs regimen is adopted to treat HCV-GT1 in recipients after liver transplantation, combination with RBV not only fails to improve the virus clearance rate and bring clinical benefits, but also increases the risk of anemia in the recipients.

Varicella(chickenpox)is an acute infectious disease with high incidence in children. It is mainly transmitted through the airborne route and vaccination is the best measure for the prevention and control. Data from post-marketing studies show the effectiveness of varicella vaccines is 80%-85%, and two-dose regimen is significantly more effective than one-dose. After inclusion of varicella vaccines into the national immunization programme, there has been a clear decrease in varicella morbidity. Despite the lack of direct evidence, there remains the risk that varicella-zoster virus might latent in the dorsal route ganglia after vaccination. Therefore, more safe and effective novel varicella vaccines are under development. This paper reviewed the progress in varicella vaccine development and their long-term efficacy and safety.

OBJECTIVE: To analysis the characteristics of sudden sensorineural hearing loss (SSHL) patients with tinnitus, and explore the relationship of characteristics of tinnitus and audiology. METHOD: Patients diagnosed as SSHL with tinnitus were studied in the research. All patients' clinical features were analyzed, such as tinnitus frequency, pure tone audiometry, tinnitus, hearing loss degree, results of residual inhibition test. RESULT: Thirty cases were identified as mild degree hearing loss, 13 cases as moderate degree, 28 cases as severe degree and 34 cases as profound degree. And hearing impaired frequency of 13 cases was ascertained at low-frequency, 39 cases at middle-high-frequency, and 53 cases at full-range-frequency. The incidence of patients with low-frequency was about 41. 9% (44/105), and it was about 21. 9% (23/105) in those with middle-frequency. And it was 36. 2% (38/105) in cases of high-frequency tinnitus. The chi-square test show statistically significant differences between patients with the low-frequency, middle-frequency and high-frequency of the hearing loss (P<0. 05). In tinnitus residual inhibition test, positive rate of convergence type masking curve was about 72.0%, tinnitus separated type masking curve 20.0%, overlapping type was 57.9%, and the spacing type was 43.5%. There was a statistically significant difference among cases with different type masking curve (P<0. 05)with the spacing residual inhibition test positive rate. CONCLUSION There are individual differences of clinical characteristics among SSHL patients with tinnitus. Tinnitus frequency is consistent with the frequency of hearing loss. Patients had the more serious the degree of hearing loss, who had more serious tinnitus grading. Cases with the converged type curve will be fit for tinnitus masking. Therefore, combining the tinnitus detection with the audiological tests, we could obtain the clinical characteristics of SSHL patients with tinnitus.
Audiometry, Pure-Tone ; Deafness ; Hearing Loss ; Hearing Loss, Sensorineural ; complications ; pathology ; Humans ; Tinnitus ; complications ; pathology

Objective:To propose a strategy for detecting early gastric cancer (EGC) under high-definition gastroscopy.Methods:Data of 469 lesions of EGC or high grade intraepithelial neoplasia (HGIN) confirmed by pathology detected at The Seventh Medical Center of Chinese People′s Liberation Army General Hospital from January 2013 to January 2020 were collected and gastroscopic images were re-interpreted. The Helicobacter pylori ( HP) infection status, lesion location in the area of atrophy or at the cardia, morphological type of lesions, lesions with/without clear or regular boundary, and lesion color were analyzed for morphological characteristics of EGC and HGIN under high-definition gastroscopy. Results:Among the 469 lesions of EGC or HGIN, HP-negative lesions accounted for 2.1% (10/469) and ulcerative lesions for 7.7% (36/469). Among non-ulcerative lesions of suspected HP infection ( n=423), there were 28 lesions in the cardia outside the atrophic area and 82.1% (23/28) were reddish under white light imaging. There were 29 non-cardiac lesions outside the atrophic area and 82.8% (24/29) were white or showed clear border under white light imaging. Inside the atrophic area, there were 73 elevated lesions, 95.9% (70/73) of which had clear border or irregular depression on the top. There were 293 flat/depressed lesions in the atrophic area, and 90.8% (266/293) had irregular border or were brown under narrow band imaging. Conclusion:According to the status of HP infection, the location and morphological category of lesions, above endoscopic features can be used as clues to detect EGC and HGIN.

Bacterial Typing Techniques ; China ; Genotype ; Humans ; Mycobacterium tuberculosis ; Tuberculosis

Objective To explore the clinical significance of colonoscopy follow-up in Chinese Lynch syndrome mismatch repair (MMR) gene mutation carriers.Methods The results of colonoscopy follow-up was analyzed in 194 MMR gene mutation carriers of 50 Lynch syndrome families.The follow-up period was from April 2001 to November 2016.The detection rates of advanced adenomas and colorectal cancers,five-year survival rate and ten year survival rate were compared between 123 patients of regular follow-up group (colonoscopy interval less than two years) and 71 patients of irregular follow-up group (time colonoscopy interval more than two years).T test,chi-square test and Kaplan-Meier method were performed for statistically analysis.Results The incidence of colorectal cancer of irregular follow up group was significantly higher than that of regular follow-up group (57.7％,41/71 vs 22.8％,28/123);and the difference was statistically significant (x2 =24.00,P＜0.01).The average age at diagnosis for colorectal cancer in irregular follow up group was younger than that of regular follow up group ((45.3 ± 1.9) years vs (48.7±1.8) years);and the difference was statistically significant (t=4.10,P＜0.01).In regular follow-up group,28.6％ (8/28) advanced-stage colorectal cancer (TNM Ⅲ or Ⅳ) was found,while in irregular follow up group,73.2 ％ (30/41) advanced-stage colorectal cancer was found,and there was statistically significant difference in pathological stage between two groups (x2 =4.90,P =0.032).The five year and ten-year survival rates of regular follow-up group were 96.2 ％ and 85.1 ％,respectively,which were both higher than those of irregular follow-up group (46.3 ％ and 28.7 ％);and the differences were statistically significant (x2 =13.20 and 14.80,both P＜0.05).The incidence of advanced adenomas of irregular follow up group was significantly higher than that of regular follow-up group (49.3％,35/71 vs 18.7％,23/123);and the difference was statistically significant (x2 =20.10,P＜0.05).The detection rate of advanced adenomas of MMR gene mutation carriers was higher than those without MMR mutation gene (85.4％,35/41 vs 14.6％,6/41);and the difference was statistically significant (x2 =5.20,P＜ 0.05).Conclusion Regular colonoscopy surveillance may decrease the incidence and mortality of colorectal cancer in MMR mutation carriers of Lynch syndrome families,and increase five-year and tenyear survival rates.

Objective To analyze the correlation of clinical phenotype and genotype and gene mutation frequency characteristics of 21-hydroxylase deficiency,and to provide the basis for clinical diagnosis and methods for early intervention.Methods The clinical phenotypic signs and examination results of 14 cases with 21-hydroxylase deficiency were collected from September 2008 to December 2016 in Children's Hospital of Shanxi Province.Point mutations,deletions and conversion mutations for gene CYP21A2 coding 21-hydroxylase were detected through using next generation sequencing(NGS) and multiplex ligation-dependent probe amplification (MLPA).The captured mutations were further confirmed with Sanger sequencing.Furthermore,the family members underwent the co-segregation validation through the Sanger sequencing or MLPA in those captured mutated sites.Results Among the total 14 cases,9 cases were identified as the salt wasting,5 cases the simple virilizing;10 cases of compound heterozygous mutations,and 4 cases of homozygous mutations.Analysis of the 14 patients revealed 8 different kinds of mutations in CYP21A2 gene.The most frequent mutations of CYP21A2 gene were I2G [50％ (14/28)] and I173N [21.4％ (6/28)],followed by Arg357Trp[10.7％ (3/28)].Del[10.7％ (3/28)] mutations including E247fs,Gly1 1 1fs and R484fs.Q319X [3.6％ (1/28)] and Arg355His[3.6％ (1/28)] were rarely found.Missense mutation was found in 10 cases,splicing mutation in 14 cases,frameshifi mutations in 3 cases,nonsense mutations in 1 case.All of the mutations were inherited from their parents,and no new mutation was found.The most common mutations for salt wasting and simple virilizing were respectively I2G[50％ (9/18)] and I173N [50％ (5/10)].Collectively,genotypes and phenotypes were matched with each other.Conclusions The combination of clinical phenotypes with laboratory examination by gene sequencing and comprehensive analysis,is helpful to early diagnosis,differential diagnosis and optimized treatment,which will improve prognosis and provide guidance for genetic consultancy.