Objective To evaluate the effects of nucleoside/nucleotide analogue treatment on immunoglobulin and complement in patients with chronic hepatitis B (CHB).MethodsA total of 157 CHB patients were recruited and divided into CHB group,liver cirrhosis (LC) group and severe hepatitis B (SHB) group.There were 50 patients who received oral antiviral treatment (lamivudine 100 mg/d,or entecavir 0.5 mg/d,or telbivudine 600 mg/d).Serum levels of complement 3 and 4 (C3,C4),C-reaction protein (CRP),hemolytic complement (CH50),immunoglobulin G,M,A (IgG,IgM,IgA),hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) were detected by enzyme-linked immunosorbent assay (ELISA) or immunoturbidimetry.Hepatitis B virus (HBV) DNA was quantified by real-time polymerase chain reaction (RT-PCR) before and 1,2,3 and 4 weeks after nucleoside antiviral therapy.Comparison of means was done by t test and Mann-Whitney test.The correlation was analyzed by Pearson correlation coefficient test.ResultsSerum IgA and IgM levels of SHB and LC patients were significantly higher than those of CHB patients (P＜0.01).Levels of C3,C4,CH50 and CRP were significantly different among three groups.Levels of C3,IgM,IgG and HBV DNA in HBeAg positive patients were significantly different from those in HBeAg negative patients.There was a statistically significant difference of IgA,IgM,C3 and CH50 levels between patients with high HBV DNA level and low HBV DNA level in HBeAg-positive patients.While in the HBeAg-negative patients,only the IgA level was significantly different with HBV DNA levels.After anti-viral treatment,immunoglobulin and HBV DNA levels were all decreased in three groups,while the serum complement level was increased compared to baseline,and the differences became significant at week 4 of treatment. HBV DNA level was negatively correlated with C3 (r=-0.78,P=0.021) and HBeAg titer was positively correlated with C3 (r=0.87,P=0.015).ConclusionsThe immunoglobulin,CRP,C3,C4,and C H50 could reflect the inflammatory activity in liver.The changes of C3 level can predict the efficacy of antiviral therapy.

Objective To investigate the susceptibility of bone marrow mesenchymal stem cell (BMSC) to hepatitis B virus (HBV) infection during induction toward hepatocyte and the role of asialoglycoprotein receptor (ASGPR) in BMSC HBV infection. Methods BMSC obtained from hepatitis B patients were tested for HBV infection and then cultured with HBV infectious serum in vitro and induced to differentiate into hepatocyte through exposure to hepatocyte growth factor (HGF), fibroblast growth factor-4(FGF-4), and epidermal growth factor(EGF). Subsequently these cells were determined for the presence of hepatitis B virus e antigen( HBeAg), hepatitis B virus surface antigen(HBsAg) and ASGPR. All experiments were repeated for 3 times in 5 different samples. The results were analyzed by non-parametric test. Results After 6 days of exposure, BMSC-derived hepatocyte-like cells expressed hepatic special genes and proteins, including alpha fetoprotein(AFP),cytokeratin18 (CK18), albumin (Alb), and manifested hepatocyte functions, including glycogen synthesis, urea secretion and albumin synthesis. Expressions of CK18 and Alb were increased, and AFP was decreased with time of induction. The BMSC were resistant to HBV infection both in vitro and in vivo or after induction toward hepatocyte. ASGPR expression level was low in BMSC, which was increased in the induced BMSC but still lower than that of the control HepG2 cells. Conclusions BMSC are resistant to HBV infection both in vitro and in vivo. The low level expression of ASGPR may be a reason for this.

Objective To construct nursing needs scale for school-age children with acute leukemia (AL) during lumbar puncture by using Delphi technique.Methods A certain number of experts were chosen for consulting to form a consulting scale.Two-round consultations were conducted and the results were analyzed.Results The effective response rate of the experts' questionnaire for 2 rounds was 100％ (17/17).The specialist group authority coefficient was 0.85.The coordination coefficients for 2 rounds were 0.437 and 0.516 respectively (P ＜ 0.01).The nursing needs scale for school-age children with AL during lumbar puncture comprised of 5 first-level indicators and 30 second-level indicators.Conclusions This scale can scientifically and quickly evaluate the nursing needs of school-age children with AL during lumbar puncture,it provides a research tool to develop individualized interventions with demand as the direction.

Objective To compare the immunoregulatory effects of Radix Millettia Speciosa (RM Speciosa) and Radix Millettia Championi (RM Championi)on immunosuppressed mice.Methods Kunming mice were randomly divided into normal control group,CTX model group,LMS positive group,RM Speciosa groups and RM Championi groups (20,10 and 5 g/kg).The mice were treated respectively with drug or NS once a day for consecutive 20 days.Mice in the other groups were injected intraperitoneally with CTX at days 8,10 and 12 to establish immunosuppressed mice model except the normal group.The changes of body weight,immune organ weight,white blood cell (WBC)number,carbon particle clearance capability of macrophages and delayed type hypersensitivity (DTH)of mice in all groups were determined and compared.Results Compared with that in CTX group,the WBC number was significantly increased (P<0.01 or P<0.05)and the degree of DTH (P<0.01) was enhanced at different doses in RM Speciosa groups and RM Championi groups.The activity of macrophages and the index of thymus and spleen were also improved in the treatment groups (P<0 .0 5 or P<0 .0 1 ).The index of thymus and the degree of DTH in RM Speciosa groups (20 and 10 g/kg)were slightly higher than those in the corresponding RM Championi groups (P<0.01 or P<0.05).For other indicators,RM Speciosa groups did not significantly differ from RM Championi groups (P>0.05).Conclusion Both RM Speciosa and RM Championi can improve the immune function of CTX-induced immunosuppressed mice,and RM Speciosa is slightly superior to RM Championi in improving specific cellular immunity.

Objective We aimed to explore the real experience of parents of children with acute leukemia during lumbar puncture,so as to provide the theoretical and practice evidence for development of individualized nursing measures.Methods Twenty parents of children with acute leukemia participated in semi-structured interviews regarding their experience of lumbar puncture.The data were colected and analyzed by the Colaizzi phenomenological methodology.Results Three themes were extracted including heavy psychological burden,lack of related knowledge of lumbar puncture and demand of multipolarity.Conclusions Attention should be paid closely to parents of children with acute leukemia during lumbar puncture.The individualized nursing measures should be taken by medical workers in order to truly meet their needs,make them stay in a good state and cooper-ate with medical workers to complete the treatment of children.

Antipsychotic Agents ; poisoning ; Clozapine ; blood ; poisoning ; Exchange Transfusion, Whole Blood ; methods ; Humans ; Infant, Newborn ; Male ; Treatment Outcome

Objective To explore the value of manganese-enhanced MRI in locating the rat visual nuclei.Methods The visual nuclei of thirty-six rats were located by 3 different ways including individual MEMRI locating (group A,n= 1 6),anatomical atlas locating (group B,n=1 6)and direct puncture by using the data obtained in MEMRI (group C,n=4).After unilateral intra-vitreal injection of MnCL2 (30 mmol/L×3 μL)in group A,the brain MRI was performed 24 h later.The location coordinate of lateral geniculate nucleus (LGN) and superior colliculus (SC)were recorded individually.The nuclei injections (3% fluorogold solution,1 μL)were performed by using different location coordinate in groups A and B.The rat’s retinas were examined under fluorescence microscope 5 days later,and the results were compared between the two groups.After brain nucleus puncture injection (30 mmol/L MnCL2 solution,0.5 μL),MRI was performed 1 h later in group C.Results The success rate was 93.8% (1 5/1 6)in group A,and 65.5% (10/1 6)in group B.The difference between groups was statistically significant (P<0.05).All the injection locations of C group were agreed with atlas.Conclusion MEMRI in the visual nucleus stereotactic can improve the accuracy of location.

The M and S segment cDNAs of hantavirus Z5 strain was amplified by RT-PCR,and the purified PCR products were cloned into vector pGEM-T and then sequenced.It was demonstrated that the M genome segment of Z5 was found to be 3 616 nucleotides in length with a single open reading frame encoding 1 135 amino acids.And the S genome segment was 1700 nucleotides in length with a single open reading frame encoding 429 amino acids.As demonstrated by the homologous analysis of nucleotides and amino acids,it was showed that the Z5 strain belonged to hantaan viruses HTN type and was the same subtype of the Z10 strain.It is conclouded that difference in nucleotide sequence exists between Z5 strain with other Hantavirus strains but high level of homology in amino acid sequences is still present.

Objective:To analyze the characteristics of clinical manifestation, auxiliary examination and gene mutation of 3-hydroxy-isobutyryl-coenzyme A hydrolase (HIBCH) deficiency to better understand this disease.Methods:The clinical manifestations and genetic results of a patient with HIBCH deficiency were analyzed. The clinical features and genetic characteristics of HIBCH deficiency were summarized based on the literature review.Results:The proband, female, one year and four months old, was admitted to Children′s Hospital Affiliated to Zhengzhou University for “vomiting and diarrhea for 15 days, dyspnea and intermittent convulsions for 13 days after digestive tract infection”. The intelligence was normal, however, the motor development was slightly delayed before onset. Physical examination showed light coma, poor response and insensitivity to light. She also had shortness of breath, weak positive three concave signs and coarse breath sound in both lungs with sputum purrs. In addition, the muscle tension of extremities was increased. Bilateral Brudzinski′s sign, Babinski′s sign and Kernig′s sign were negative. Serum hydroxybutyryl carnitine (C4OH) was increased. Cranial magnetic resonance imaging (MRI) showed atrophy in bilateral cerebral hemispheres and abnormal symmetry signals in bilateral globus pallidus and cerebral peduncle. Novel compound heterozygous variants of HIBCH, c.489T>A (p. C163*) and c.740A>G (p. Y247C), were found in the patient, which respectively inherited from her healthy parents. Her symptoms were relieved after“cocktail”therapy and symptomatic treatment. Literature related to HIBCH deficiency published all around the world was reviewed. As a result, 17 articles, including 24 cases, had been reported. The majority of patients presented with poor feeding, dystonia and progressive motor developmental delay in early infancy. Cranial MRI showed lesions in bilateral basal ganglia. Serum C4OH concentration was elevated. And compound heterozygous or homozygous variants of HIBCH gene were found in patients with HIBCH deficiency.Conclusions:The detection of serum amino acids and acylcarnitine profiles on HIBCH deficiency was relatively specific and it was helpful to make a clear diagnosis by combining with cranial MRI and genetic tests. In this study, a case of HIBCH deficiency was confirmed, which expanded the mutation spectrum of HIBCH gene. Meanwhile, summarizing the clinical and genetic characteristics of cases reported improved understanding of HIBCH deficiency.

Objective To compare the immune effects of Coxsackievirus B3 (CVB3) capsid protein VP1 expressed bacterially, recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1which express VP1 protein in mice. Methods After expressed in prokaryotic cells, VP1 protein was purified. Recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1 were amplified and extracted. Six to 8-week-old, male BALB/c mice were divided into four groups randomly. Each group contained 18 mice. The mice of pcDNA3/VP1 group or VP1 protein group were immunized intramuscularly with three injections at three weeks apart, of recombinant plasmid pcDNA3/VP1 at a dose of 100 μg/mouse or recombinant protein VP1 at a dose of 50 μg/mouse. The mice of rAd/VP1 group were immunized intramuscularly twice at two weeks interval with rAd/VP1 at a dose of 1.2 × 107 PFU. The control group was mock-immunized with 100 μl of PBS intramuscularly. Mice were bled from the retroorbital sinus plexus every two weeks after each immunization. ELISA and micro-neutralization test were used to detect levels of CVB3-specific IgG antibody and neutralizing antibody titers in the sera of immunized mice. Three weeks after the last immunization, the cytotoxic T lymphocyte(CTL) killing activity of spleen lymphocytes was detected with CCK-8 assay. Subsequently, virus titers in the sera of immunized mice were determined by the 50% cell culture infective dose( CCID50 ) assay on HeLa cell monolayers and percentage of animals surviving were observed after lethal CVB3 attack over a period of 21 days. Results The titers of specific IgG antibody and neutralizing antibody in sera of VP1 protein immunized mice were higher than other groups( P ＜0.05 ). While CTL killing activity of spleen lymphocytes of VP1 protein immunized mice was lower than mice in rAd/VP1 group( P ＜0. 05). Virus titers in sera of VP1 protein immunized mice were lower than the mice in pcDNA3/VP1 or rAd/VP1 groups ( P ＜ 0.05 ), while survival rate was significantly higher than these two groups ( P ＜ 0.05 ).Conclusion VP1 protein induced higher level of humoral immune response and acquired obvious immune protection effects in mice. The immunizing potency of VP1 protein vaccine surpassed plasmid pcDNA3/VP1or recombinant adenovirus rAd/VP1. It appeared to be a promising candidate among the three different vaccines.