Objective To observe the changes of CD_(28) and soluble CD_(27)(sCD_(27)) among HIV-1-infected Chinese patients receiving highly active antiretroviral therapy(HAART) and to explore their clinical significance.Methods The HIV-RNA viral load(VL) and cell counts of CD_3~+CD_4~+,CD_3~+CD_8~+,CD_4~+CD_(45)RA~+CD_(62)L~+,CD_4~+CD_(28)~+,CD_8~+CD_(28)~+ and the content of sCD_(27) were determined in 28 cases of HIV-1-infected patients at the baseline and 12,24 and 48 weeks respectively after the treatment with three drug regimens based on two nucleoside reverse transcriptase inhibitors(NRTIs) and an Non-NRTI(NNRTI) for 1 year.Results The average of HIV-RNA VL before the treatment was 5.25 logs,and decreased to 3.30,1.92 and 1.74 logs after the treatment for 12,24 and 48 weeks respectively.The cell counts of CD_3~+CD_4~+,CD_3~+CD_8~+,CD_4~+CD_(45)RA~+62L~+,CD_4~+CD_(28)~+,CD_8~+CD_(28)~+ before the treatment were(236?85),(917?345),(58?15),(175?63) and(285?87) cells/?l,and changed into(327?112),(803?343),(114?25),(286?132) and(387?201) cells/?l after the treatment for 48 weeks respectively.The content of sCD_(27) decreased from(475?123) U/ml to(174?77) U/ml after the treatment for 48 weeks.The HIV RNA VL had significantly negative correlation with cell counts of CD_4~+T,CD_4~+CD_(28)~+ and CD_8~+CD_(28)~+,and there was positive correlation between HIV RNA VL and the level of sCD_(27).Conclusion The immune reconstitution and activation depend on the efficacy of antiretroviral therapy.The HIV RNA VL and cell counts of CD_4~+T are the representative parameters of immunologic and viral responses of HAART.The cell counts of CD_4~+CD_(28)~+ and CD_8~+CD_(28)~+ and their percentages show concomitant changes with viral load,and play an effective role in the progression and prognosis of AIDS.The level of sCD_(27) is positively correlated with HIV RNA VL and negatively with counts of CD_4~+T cell.The detection of sCD_(27) by enzyme linked immunosorbent assay(ELISA),which is much more convenient and cheaper than RT-PCR and flow cytometry,may serve as an important parameter to judge the pathogenetic condition of HIV-1-infected patients and to examine the therapeutic effects of HAART in the medical resource-limited area.

objective Microbial limit test(MLT)for Dahuang Zhechong pill was established and verified to supply scientific foundation for the establishment of MLT for antibacterial and anti-inflammatory Chinese patent medicine,and to supply principle guidance for the verification of the method issued in Chinese pharmacopoeia(2005 edition);meanwhile the antibiogram of Dahuang Zhechong pill was probed,which can provide evidence for its clinical administration.Methods With centrifugal sedimentation method and culture medium diluted method,the recovery test was done on each tested bacterial strain to validate the accuracy and feasibility of MLT.Results Dahuang Zhechong pill had a stronger inhibitory effect on staphylococcus aureus and bacillus subtilis,had a weak inhibitory effect on escherichia coli,and had no effect on candida albicans and aspergillus niger,indicating that the antimicrobial spectrum of Dahuang Zhechong pill is Gram-positive bacteria.Conclusion Centrifugal sedimentation method combined with culture medium diluted method can eliminate the anti-bacterial effect of Dahuang Zhechong pill during the experiment,thus the microorganisms in the polluted pill can be detected successfully.

Objective To study the association between house dust endotoxin levels and the airway responsiveness as well as allergen sensitization in adolescence in the urban and rural areas of Guangdong province.Methods Middle school students,aged 13-14,188 (male 109,female 79) from Guangzhou,129(male 70,female 59) from Conghua with positive responses,and 231 (male 125,female 106) and 307(male 145,female 162) of their respective reference students are selected from questionnaire prescreening.A detailed questionnaire,the lung function test,histamine bronchial provocation test,skin prick test and peripheral blood eosinophil count are undertaken.Seventy-six students from Guangzhou and 80 from Conghua were selected randomly for mattress dust sample collection to detect the levels of endotoxin.Results The prevalence of wheeze symptom,diagnosed asthma,nasal symptoms and allergic rhinitis of Guangzhou was significantly higher than that of Conghua (P＜0.0001),with 28.6％,27.7％,66.0％,46.4％ for Guangzhou respectively,and 5.0％,2.5％,31.0％,6.2％ for Conghua respectively.The positive rates of bronchial provocation test and skin sensitization against Dermatophagoides pteronyssinus(Der.p),D.farinae (Der.f),cat and dog allergen in the students of Guangzhou were significantly higher than that of Conghua (P＜0.0001).The positive rate of cockroach of Conghua was significantly higher than that of Guangzhou (P＜0.0001).The levels of endotoxin (amount,concentration and load) of samples from Conghua were significantly higher than that from Guangzhou (Conghua:6452 EU,10.95 EU/mg,1794 EU/m2 ;Guangzhou:1591 EU,6.45 EU/mg,508.80 EU/m2,respectively).PD20-FEV1 was correlated positively with endotoxin amount(r=0.174,P＜0.05).The diameters of the skin wheal size for Der.p and Der.f were correlated inversely with endotoxin amount and load (P＜0.001).Conclusion The prevalence of wheeze symptom,diagnosed asthma,nasal symptoms and allergic rhinitis in 13-14 year-old adolescent of Guangzhou was significantly higher than that of Conghua.Cockroach was the second major allergen of Conghua followed by house dust mite.The level of endotoxin was correlated positively with PD20-FEV1,but correlated inversely with skin sensitization to Def.p and Der.f.The study suggests that environmental endotoxin may have effect to protect children from developing allergic diseases.

Objective To change methods of experiments for increasing Light Cycler's ability to seek out samples of function.Methods To increase the quantity of the reaction of substance,proceed to amplification in advance,and increase the seeking temperature etc.Results Above methods affects inapparent to the accuracy of the examination result,the method of increasing the seeking temperature is particularly simple and practical.Conclusion The inside clinical laboratory science process can adopt the above methods to increase the LightCycler's seeking-out function according to actual circumstance.

Purpurin is a common component ofRubia cordifolia L. The study on its molecular target was useful for elucidating the therapeutic material basis and action mechanism ofR. cordifolia. HT-29 cells were used in the cell culture. The highly expressed G Protein-coupled Receptor-35 (GPR35) agonist was used as target. The label-free optical biosensor cellular assay was used to investigate the agonist activity ofpurpurin at an endogenous receptor. The results showed thatpurpurin can cause DMR response in HT-29 cells. And the DMR response curve type was consistent with zaprinast. Its EC50 was 6.142± 0.189μmol·L-1. In addition,purpurinhad desensitization effect on GPR35 agonist zaprinast in HT-29 cells. GPR35 agonist ML145 blocked the DMR ofpurpurin. It was concluded thatpurpurinwas the GPR35 agonist.

Aim To investigate the axonal regeneration effect of salidroside in MCAO rats and its potential mechanism.Methods Thirty-six healthy adult male Sprague-Dawley rats were randomly divided into three groups: sham, MCAO, MCAO+Sal groups.The rats were subjected to focal cerebral ischemia/reperfusion with suture-occluded method.Neurological deficit testing was performed with Zea Longa scale.The protein expression of p-Akt(Ser473), Akt, p-GSK-3β(Ser9), GSK-3β, p-CRMP-2(Thr514) and CRMP-2 in side cerebral ischemic tissues were determined using Western blot analysis.NF200 immunofluorescence staining was used to evaluate axonal regeneration.Results Compared with MCAO group,salidroside significantly improved the neurological deficit,up-regulated the protein expression of NF200,p-Akt and p-GSK-3β,and inhibited the protein expression of p-CRMP-2.Conclusions Salidroside improves neurological function recovery after focal cerebral/ischemic injury in rats,which may be associated with the up-regulation of phosphorylated Akt and GSK-3β and inhibition of phosphorylated CRMP-2,thereby promoting axonal regeneration.

Objective To evaluate the effects of sub-acute alcoholism on cardiac structure and function, and investigate the mechanisms of aldehyde dehydrogenases-2 (ALDH2)alleviating the damage of heart caused by acute myocardial infarction.Methods The wild mice with ALDH2 (+/+) (WT group) and mice with knockout type ALDH2 (-/-) genotypes (KO group) were raised and then divided into four groups according to the presence or absence of sub-acute alcoholism: WT group (n=10), KO group (n=16), WT+alcoholism group(WT+E,n=10) and KO+alcoholism group(KO+E,n=10).The mice of WT+E group and KO+E group were fed with high-dose of ethanol(2 g/kg per day for 8 days), while the mice of WT group and KO group were treated with equal amount of saline instead.Acute myocardial infarction models were established in all mice after ethanol administration,and blood ethanol concentration, cardiac function, myocardial infarct size, the activity of ALDH2, and the key molecules of PI3K/Akt signal pathway and caspase-3 mRNA were detected one week after modeling.Statistical analysis was performed using SPSS 17.0.Differences in levels of detected biomarkers between groups were assessed using Chi-squared or One way ANOVA, and P<0.05 was considered to be statistically significant.Results (1) The mortality rates of WT group, KO group, WT+E group and KO+E group were 20.0%, 30.0%, 31.3% and 37.5%, respectively.(2)Compared with WT group and KO group, the blood ethanol concentration was higher and the damage of liver was more severe in WT+E group and KO+E group(P<0.05).(3)The fraction shortening of short axis of left ventricle(FS) and left ventricular ejection fraction were higher in WT group and WT+E group compared with KO group and KO+E group(P<0.05).(4) The area of myocardial infarction was largest in KO+E group, followed by KO group, WT+E group, and WT group (all P<0.05).(5) The activity of ALDH2 in WT group was higher than that in other groups, and the ALDH2 activity in KO+E group was lower than that in KO group (P<0.05).(6) There was no significant difference in expressions of PI3K among four groups.The level of p-Akt was highest in WT group, followed by WT+E group, KO group, and KO+E group (all P<0.05).The levels of caspase-3 mRNA was highest in KO+E group, followed by KO group, WT+E group, and WT group (all P<0.05).Conclusions Myocardial damage caused by acute myocardial infarction can be aggravated by sub-acute alcoholism, while ALDH2 protection can effectively alleviate the damage effects of sub-acute alcoholism on myocardial infarction.The mechanism of protective effects of ALDH2 on acute myocardial infarction may be related to attenuation of cardiocytes apoptosis mediated by PI3K/Akt signal pathway.

BACKGROUNDThe bovine herpersvirus structural protein BVP22 exhibits the remarkable property of intercellular trafficking whereby the protein fused to BVP22 spreads from the cell in which it is synthesized to surrounding cells. This function of BVP22 might be exploited to overcome the low efficiency of genes and gene products delivery, which is a major hurdle in gene therapy. The aim of this study is to investigate the cellular localization and intercellular trafficking of BVP22 in vitro and in vivo and provide scientific data for its application in gene therapy of human lung cancer.

METHODS801D cells were transfected respectively with plasmids pEYFP and pEYFP-BVP22 mediated by Lipofectamin and were selected by G418 to establish clone cell lines. The expression and cellular localization of BVP22 were examined by direct observation of YFP. Intercellular trafficking of BVP22 in vitro was detected by fluorescence-activated cell sorting (FACS) and immunocytochemical staining with YFP-antibody. Subcutaneous 801D tumors in nude mice were established to investigate the intercellular trafficking of BVP22 in vivo.

RESULTSClone cell lines pEYFP-801D and pEYFP-BVP22-801D were established successfully. Cellular localization of BVP22 displayed heterogenic- ity. BVP22 was present in nuclei in most cells and only a few cells showed filamentous cytoplasm pattern. The results of FACS showed that the ratios of YFP-positive cells in mixed cells did not enhanced significantly. Immunocytochemical staining demonstrated that the nuclei of almost all cells were stained positively after pEYFP-BVP22-801D and 801D were cultured for 24h. Intercellular trafficking of YFP-BVP22 could be observed in subcutaneous 801D tumors in nude mice by immunohistochemical staining.

CONCLUSIONSBVP22 displays nuclear localization in most cells. Its nuclear localization might be related to cell mitosis and intercellular trafficking. BVP22 can mediate intercellular trafficking of fusion protein in vitro and in vivo.

Brucellosis is far more frequent in a pasturing area in the northern part of our country and it has many clinical manifestations. It may cause multiple organ damage and its features lack specificity. It is rare in the south, so it is extremely easy to be misdiagnosed or overlooked. The retrospective analysis of a case with Brucellosis misdiagnosed as osteoarthrosis provides a guide for clinical doctors to understand Brucellosis, so that early diagnosis would be accessible, and prognosis could be improved.
Brucellosis ; diagnosis ; Diagnostic Errors ; Humans ; Osteoarthritis ; Retrospective Studies

Parkinson's drug-induced dyskinesia is a common complication after levodopa treatment,and its clinical manifestations are complex.A case of Parkinson's L-dopa-induced dyskinesia misdiagnosed as epilepsy in our hospital is reported.The patient,a 69-year-old male,was hospitalized on May 2,2018 due to "15 years of retardation,recurrent limb twitching with breathing for more than 6 months,sudden loss of consciousess for 10 minutes".The patient took madopar for a long time and had obvious drug complications.In July 2017,the use of rasagiline improved manifestations slightly.In November 2017,repeated limb twitches began to occur.The onset time ranged from several minutes to one hour,and there was a gradual occurrence of twitching accompanied by obvious breathing.On May 1,2018,the patient was sent to our hospital for emergency treatment after limb twitching again.Due to the long-term history of hormone use,and tranquilizers and antiepileptic drugs were used to control the condition of the patient,the patient was misdiagnosed as epilepsy at first.With the adjustment of drugs and the change of condition,the patient was finally diagnosed as Parkinson's L-dopa-induced dyskinesia.The clinical manifestations of the disease include peak dyskinesia,bipolar dyskinesia and dystonia.The pathogenesis of Parkinson's disease is not completely clear and its clinical manifestations are diverse.However,there are few reports of Parkinson's disease,whose manifestation is loss of consciousness due to paroxysmal breath holding and hypoxemia.Therefore,in order to diagnose epilepsy in patients with Parkinson's disease,the possibility of dyskinesia should be excluded first.Therefore,the rational use of drugs is particularly important in the middle and late period of Parkinson's disease.