The high mutation rate during hepatitis B virus (HBV)replication leads to HBV quasispecies.The study of HBV quasispecies has an important significance for the hepatitis B pathogenesis,prognosis,and outcome prediction.Recently,the next-generation sequen-cing (NGS)is extensively used in many biological and medical fields due to its high throughput,ultra-deep coverage,and high sensitivity, which also brings new strategies to HBV quasispecies studies.This article describes the working principles and features of several commonly used NGS technologies,reviews the application of NGS technologies in HBV quasispecies detection in recent years,and particularly discus-ses the variations in different regions of HBV genome and the population characteristics of HBV quasispecies.For now,NGS technologies used in HBV variation detection mainly presents the advantages of high sensitivity and high throughput.However,how to lower the cost,in-crease the accuracy of sequencing,and standardize the procedure of massive sequencing data management and personalized analysis are still challenging problems.The issue about NGS that matters at present and urgently needs to be solved is how to overcome its limitations,and then put it into HBV-related studies and ultimately clinical application.

Objective To evaluate methodologically particle‐enhanced turbidimetric immunoassay (PETIA ) for determining quantitatively Cystatin C .Methods Cystatin C was determined quantitatively with PETIA by Cobas 8000 automatic biochemical analyzer ,and imprecision ,veracity ,sensitivity ,linearity range of the method were investigated .The correlation of fresh serum sam‐ple results between PENIA and PETIA was analyzed .The reference interval was verified .Results The within‐run CV imprecision of samples with low and high concentration of adiponectin was 3 .67% and 1 .15% respectively ,the between‐run CV imprecision of samples with low and high concectration of adiponectin was 4 .08% and 1 .53% respectively .The accuracy of the measurement devi‐ation was only -1 .25% .The sensitivity was 0 .07 mg/L .When the adiponectin concentration was between 0 .2 -8 .0 mg/L ,the method showed good linearity .A strong correlation between PETIA and PENIA was confirmed (Y = 0 .945 8X+ 0 .048 6 ,r2 =0 .991 3 ,r=0 .995 6) .Among 40 cases of healthy physical examination sample ,2 cases were with cystatin C level out of the refer‐ence range supplied by reagent kit .The values that were more than 90% in validation interval .Conclusion The PETIA assay is a convenient ,rapid ,cheap ,accurate and reliable method for quantitating the human serum concentration of adiponectin .This assay is suitable for clinical routine analysis .

Polycyclic aromatic hydrocarbons(PAHs) is a class of organic pollutants widely distributed in various environmental medium,and human exposure can occur by many routes (inhalation,ingestion,and skin contact),regardless of the source. PAHs can seriously impact human health and environment due to their high toxicity,stability. Studies on the toxicity of PAHs have involved in many fields,such as genotoxicity,hepatotoxicity,developmental toxicity,carcinogenesis and so on. The development of studies on genotoxicity of PAHs was reviewed in this paper,including DNA damage,chromosome damage and genetic susceptibility.

OBJECTIVE To investigate systematically the existence of bactericidal/permeability increasing protein(BPI) in normal rat organs and tissues,for providing important information about BPI in clinical application.METHODS The total RNA was extracted from organs and tissues homogenates.Then the first-strand cDNA was synthesized and BPI DNA was amplified by PCR.RESULTS BPI mRNA was found in kidneys,ovary,testes,liver,small intestine,large intestine,thymus,spleen and neutrophils of 21 various organs and tissues.Among them,BPI mRNA content was the richest in testes,relatively rich in liver and large intestine.CONCLUSIONS BPI mRNA presents in many organs and tissues of rat and it indicates that the role of BPI in host defense against bacterial infection is relatively widespread.

Objective To analyze the changes of the quality of life of patients with advanced lung cancer accompanied with dyspnea before and af?ter methylprednisolone therapy,so as to investigate the effects of methylprednisolone on the quality of life of the patients. Methods Totally 134 ad?vanced lung cancer patients with dyspnea were divided into two groups,the methylprednisolone group and the control group,according to whether they consented to the use of hormone therapy. All of them were followed with visual analog scale(VAS),the quality of life questionnaire?core 30 (EORTC QLQ?C30)and the quality of life questionnaire?lung cancer module 13(QLQ?LC13)questionnaire. Results At one hour after treatment compared to before treatment,the VAS scores for both groups were significantly decreased(P<0.01),and the score for the methylprednisolone group were lower than the control group(P<0.05). At two weeks after treatment compared to before treatment,social function,emotional function, and overall quality of life scores for both groups were decreased(P<0.05). But the scores of pain,dyspnea,insomnia,loss of appetite,constipation, dysphagia were increased(P<0.05). Compared to the methylprednisolone group,the dyspnea and pain score for the control group was higher(P<0.05). Conclusion Methylprednisolone may improve the quality of life of advanced lung cancer patients with dyspnea in short term and slow down the deteriorating process of their quality of life.

OBJECTIVE:To determine plasma concentration of caffeine,dapsone and chlorzoxazone in rats,and to calculate pharmacokinetic parameters. METHODS:6 rats were given the mixture of caffeine,dapsone and chlorzoxazone intragastrically, 1.5,2 and 3 mg/kg,respectively. 0.2-0.3 ml blood were collected before medication and 0.5,1,2,3,4,6,8,12,24 h after medication.The plasma sample was treated with solid phase extraction. The plasma concentration of caffeine,dapsone and chlorzoxa-zone were determined by HPLC using N-(2-Hydroxyethyl) phthalimide as internal standard. The pharmacokinetic parameters were calculated using DAS 2.0 software. RESULTS:The linear ranges of caffeine,dapsone and chlorzoxazone were all 0.2-30 μg/ml (r were 0.996 4,0.996 1,0.998 8,respectively). The limit of quantitation were 0.2 μg/ml. The recoveries of low-concentration, medium-concentration and high concentration were(84.8±3.6)%-(111.4±10.2)%(RSD were 4.3%-9.8%,n=3),(107.0±13.3)%-(113.5±8.1)%(RSD were 7.1%-14.0%,n=3),(104.2±10.8)%-(111.1±12.2)%(RSD were 8.0%-11.0%,n=3). Pharmacoki-netic parameters were as follows as tmax(1.70±0.99),(1.50±1.00),(1.92±0.80)h;t1/2(0.73±0.22),(2.77±1.35),(2.78±2.34) h;cmax (2.60 ± 0.50),(5.78 ± 1.19),(9.76 ± 1.37) mg/L;AUC0-t (8.43 ± 0.79),(20.68 ± 1.91),(26.71 ± 2.45) mg·h/L(n=6). CONCLUSIONS:The method is simple,sensitive and accurate,and can be used for the plasma concentration determination and pharmacokinetic study of caffeine,dapson and chlorzoxazone.

Hepatitis B virus ; classification ; genetics ; High-Throughput Nucleotide Sequencing

Objective To observe the clinical efficacy of naloxone plus aminophylline in the treatment of children with acute respiratory failure.Methods 72 patients with acute respiratory failure were randomly divided into the study group and the control group.36 cases in the study group were given naloxone plus aminophylline therapy, 36 cases of the control group received conventional therapy.And the clinical efficacy was compared.Results The total effective rate of the study group was 91.67%,which was significantly higher than 66.67% of the control group (χ2 =6.82,P <0.05).In the comparison of the effect of improving blood indicators,arterial blood pressure of the study group was (67.51 ±4.11)mmHg,which was significantly higher than (61.03 ±4.08)mmHg in the control group (t =2.64,P <0.05).The oxygen saturation of the study group was (93.55 ±8.05)%,which was significantly higher than (79.62 ±10.22)% of the control group (t =2.29,P <0.05).The arterial carbon dioxide partial pres-sure of the study group was (69.03 ±5.71)mmHg,which was also higher than (61.52 ±4.09)mmHg of the control group (t =2.22,P <0.05).The incidence rate of adverse reactions of study group was 2.78%,which was lower than 22.22% in the control group (χ2 =6.22,P <0.05).Conclusion Naloxone plus aminophy -lline used in children with acute respiratory failure obtain the desired therapeutic effect,not only can effectively improve blood indicators of children,and without significant adverse reactions,drug safety is high,it is worthy of promoting.

Objective: To summarize surgical result of single ventricle with total anomalous pulmonary venous connection (TAPVC) in relevant patients.
Methods:A total of 25 patients of single ventricle with TAPVC treated in our hospital from 2008-09 to 2016-03 were summarized. The patients were between 5 days to 14 years of age with body weight at (3.3-49.5) kg. There were 22 patients with supracardiac type TAPVC, 1 with infracardiac type TAPVC and 2 with mixed type TAPVC;in addition,17 cases having complete atrio-ventricular canal defect, 2 having tricuspid atresia, 4 having more than moderate atrio-ventricular valve regurgitation, 5 having pulmonary atresia and 9having pulmonary venous obstruction.
Results:There were 5/25 (20%) patients died at the early stage for low cardiac output syndrome. Followed-up study was conducted in 20 survivals for (1-65) months and 7 patients had re-surgery including 5 with modified Fontan operation, 1 with atrio-ventricular valve replacement due to severe atrio-ventricular valve regurgitation, 1 with pulmonary venous stenosis repair+atrio-ventricular valve replacement due to pulmonary venousobstruction and severe atrio-ventricular valve regurgitation. 3 patients died during followed-up period because of low cardiac output syndrome, protein lost intestine disease and neurological injury respectively. The rest survivals had good general condition, no residualpulmonary venous obstruction was found.
Conclusion: The patients of single ventricle with TAPVC were frequently associated with right atrial isomerism and complex cardiac anomaly;surgical treatment still having the high mortality.

BACKGROUND:Occipitocervical fusion is a major method for malformation of craniocervical junction. In patients without osteoporosis, the degeneration of cervical vertebra mainly presents in the intervertebral disk. The height of the vertebral body is constant basical y. Thus, the ratio (S value) of the height of cervical disc and the height of cervical vertebra can be used to measure the degeneration of cervical vertebra. The smal S value indicates severe degeneration of cervical vertebra.
OBJECTIVE:To measure the lateral radiograph of cervical vertebra in patients with craniocervical malformation undergoing occipitocervical fusion, to analyze the relationship between occipitocervical fixed angle during fusion and lower cervical spine degeneration after fusion, and to identify an optimal angle of occipitocervical fusion.
METHODS:A total of 21 patients with craniocervical malformation undergoing occipitocervical fusion were included. According to the occipitocervical angle (0c-C2 angle) immediately after fusion, the patients with craniocervical malformation undergoing occipitocervical fusion were assigned to three groups:occipitocervical angle 9°-22° group, occipitocervical angle<9° group, and occipitocervical angle>22° group. Immediate postoperative 0c-C2 angle in 9°-22° belonged to the normal angle range. S value and JOA score in each group were measured before and after fusion, during final fol ow-up. The statistics were compared.
RESULTS AND CONCLUSION:JOA scores in the occipitocervical angle 9°-22° group, occipitocervical angle<9° group, and occipitocervical angle>22° group, were respectively, (7.3±1.7) points, (7.2±1.6) points, and (7.3±1.5) points, before fusion, and (14.2±1.5) points, (13.5±1.6) points and (13.3±1.5) points after fusion. JOA scores were improved significantly in the three groups. JOA improvement was significantly better in the occipitocervical angle 9°-22° group than that in the occipitocervical angle<9° and>22° groups. Preoperative S values were respectively 0.440±0.017, 0.441±0.016, and 0.440±0.018 in the occipitocervical angle 9°-22° group, occipitocervical angle<9° group, and occipitocervical angle>22° group, and no significant difference was detected among the three groups. No significant difference in S value was detectable in the occipitocervical angle 9°-22° group between postoperative final fol ow-up and pre-operation. The S value was significantly smal er at postoperative final fol ow-up than pre-operation in the occipitocervical angle<9° and>22° groups. These results indicated that during occipitocervical fusion, occipitocervical angle should try to be normal, more than or less than normal range wil accelerate the degeneration of lower cervical spine.