Hepatitis B virus exists in the form of quasispecies.Quasispecies are an important basis for the pathogenesis and drug resistance of hepatitis B virus.Researches for quasispecies could contribute to evaluating the progress of disease, predicting drug resistance, adjusting the therapeutic regimen and performing individual treatment.

Long-term use of nucleoside ( nucleotide ) analogues for anti-HBV treatment care easily lead to drug resistance. As compared to genotypic resistance detection, the detection technology of phenotypic resistance is a method used to analysis the sensitivity of antiviral drugs in vitro cell model, the result can directly reflect the relationship between drug-resistance mutations and drug susceptibility, thus perform a better prediction of clinical resistance.Among those technologies, some analysis strategies like transient transfection assay, stable cell lines and enzymatic assay, have been developed for early diagnoses as well as dynamic monitoring of drug resistance, and contribute greatly to proper selection of therapeutic regimen, reduction of resistance rate and implementation of individual treatment.

With the rapid advancement of immunology and molecular biology,many new revelations and ideas were drew for clinical immunology in laboratory medicine.In the current circumstances of application of clinical pathway management and individual medical care,how to accurately grasp these changes to build perfect quality system for clinical immunology laboratory,clarify the immunology laboratory pathway and integrate that into clinical pathway management,make solid progress for the promotion and application of individual diagnosis and evaluate the diagnostic value of new technique and item for clinical immunology laboratory accurately might be worth pondering for every people working in clinical immunology laboratory.In this paper,the status quo and reflection of clinical immunology is reviewed.

Drug-resistance mutation is inevitable during long-term monotherapy with nucleus(t)ide analogues in hepatitis B patients.Early detection the drug-resistance mutations could contribute to adjusting the therapeutic regimen,reducing resistance rate,improving therapeutic efficacy and performing individual treatment.

With the development of molecular biology and molecular genetics,more attention has been taken to the advanced techniques which can be used to detect minority mutations.In recent years,many techniques such as amplification refractory mutation system,PCR clamping,pyrophosphorolysis-activated polymerization,co-amplification at lower denaturation temperature-PCR and digital PCR have been developed,which will make the hereditary disease diagnosis,cancer detection,noninvasive antenatal diagnosis step into a new stage in the near future.This review presents an overview of the principles,aoolications,advantages and disadvantages of these techniques.

Objective To establish a new approach for quantitative detection of VAD1 mRNA in cryptococcus neoformans by RT-FQ-PCR, and evaluate the treatment efficacy of CNM. MethodsThe primers and TaqMan probe were designed according to the published sequence of VAD1 mRNA (GenBank),and RT-FQ-PCR method to detect VAD1 mRNA was established. Cerebrospinal fluid from 25 CNM patients and 30 controls were detected and sensitivity and specificity of the method were evaluated. VAD1 mRNA concentration in cerebrospinal fluid from both acute phase and recovery phase of 25 CNM patients were also detected and significance of CNM treatment efficacy with VAD1 mRNA analysis was evaluated. Results Correlation coefficient of standard curve was - 0. 997 9 in detection of VAD1 mRNA by RT-FQ-PCR, and the detection limit was 101 copies/μl. The intra CV of plasmid standard for high, medium and low concentrations were 0. 65% ,0. 89% and 1.23% respectively, the sensitivity of cryptococcus neoformans detection by RT-FQPCR was 96% (24/25) ,while specificity was 100% (30/30). VAD1 mRNA concentration in acute phase were significant higher than that in recovery phase (3. 042 ±0. 906 vs 2. 187 ±0. 665 ,t =4. 583 ,P ＜0. 01).Conclusions The established RT-FQ-PCR method for the detection of VAD1 mRNA is provided with sound sensitivity, specificity and reproducibility, which might be fit for the detection of VAD1 mRNA. The expression level of VAD1 mRNA is relevant with the treatment efficacy of CNM.

Despite decades of the prevention and control.China is still one of the countries with high prevalence of hepatitis B virus (HBV).It is imperative for us to innovate.Lab diagnostic platforms for drug-resistant mutation and HBV genotyping to enhance the management level in patients with CHB.Specifically,the following three strategies should be included:firstly,choose an appropriate platform for drug-resistant mutation detection according to the actual situation in laboratory.Secondly,establish a practical and simple genotyping platform for real-time monitoring of HBV genotypes.Thirdly,combined with clinical,explore the correlation among genotypic resistance,phenotypic resistance and clinical resistance when different medications are adopted,confirm the percentage or threshold of resistant strains causing clinical resistance,and ascertain the influence of HBV genome polymorphism (genotype) on clinical prognosis.These strategies above mentioned will lay a foundation for CHB personalized treatment and prognosis.

Objective To investigate the effect of melatonin(MT)on tau hyperphosphorylation in hippocampus of mice with exposure to lead(Pb)at early development stage and its possible mechanism.Methods Healthy C57BL/6 mice were randomly divided into four groups:control group(received normal water),lead exposure group(exposed to 0.2% Pb acetate from postnatal day 1(PND 1)to PND 21 through drinking water),MT group(received 50 mg/mL MT through drinking water since 12-month-old for 3 months)and MT combined with lead exposure group(exposed to 0.2% Pb acetate from PND 1 to PND 21 and then given 50 mg/mL MT since 12-month-old for 3 months through drinking water).The Pb levels in the blood and hippocampus were determined by graphite furnace atomic absorption spectrometry.Morris water maze was used to detect the spatial learning and memory.The phosphorylation level of tau and the protein level of GRP78 and CHOP in hippocampus were detected by Western blotting.Results Compared with the control group,the Pb levels in the blood and hippocampus were significantly increased in lead exposure group(P<0.05),while MT treatment did not affect the Pb levels both in the blood and hippocampus.In the lead exposure group,the phosphorylation level of tau in the hippocampus was significantly increased compared with control group(P<0.01),and after treatment with MT,the phosphorylation level of tau in MT combined with lead exposure group was significantly decreased compared with the lead exposure group(P<0.05).Furthermore,the expression of GRP78 and CHOP in the hippocampus was significantly higher in lead exposure group than in control group(P<0.05),and after treatment with MT,the expression levels of GRP78 and CHOP were lower than those of the control group.Conclusion MT can ameliorate the phosphorylation level of tau in the hippocampus and defects of learning and memory in C57BL/6 mice with exposure to lead at early development stage,and this may be related with the modulation of endoplasmic reticulum stress.

Objective To explore the role of sCD44v6 in the diagnosis of colorectum cancer. Methods ELISA was used to detect the concentration of sCD44v6 of 81 patients after or before surgery.Results The level of sCD44v6 in the group of colorectum cancer was 200.5?31.1 ng/ml, which was higher than that of normal controls (45.6?9.5ng/ml) and that of patients with coloritis (48.3?7.6ng/ml). The concentration of sCD44v6 in the group of colorectum cancer with metastasis(280.1?26.9ng/ml) was higher than that of the group without metastasis(160.9?34.4ng/ml). The level of sCD44v6 was related to tumor burden.Conclusions The level of sCD44v6 is closely related to colorectal cancer and its metastasis. It would be useful in the diagnosis, prognosis, and forcasting of metastasis.

Innate immunity initially resists the infection of pathogenic microorganism in host immune response.Recent researches confirmed that mitochondria participated in a wide range of innate immune pathways,mainly including contributing to innate immune activation,regulating antiviral signaling pathways and antibacterial immunity.Therefore,further studies on the relationship among mitochondria,hepatitis B virus(HBV)infection and innate immune response might contribute to elucidate the mechanism of chronic HBV infection and explore the mechanism of host immune to clear HBV.Here,mitochondria playing a vital role in regulations of innate immune response,HBV infection tending to chronicity by suppressing innate immune response and chronic HBV infection by regulating the innate immune response through injuring mitochondria,were reviewed.