Strain NCIMB 10467, a lignin degrader, was reclassified as genus Burkholderia according to its 16S rDNA sequence. It seems that the metabolism of protocatechuate by this strain is diverse under the induction of various substrates. A 9505-bp DNA fragment extending from α conserved region of the gene, which encodes β subunit of orthocleavage protocatechuate 3, 4-dioxygenase(P34D; EC 1.13.11.3),was obtained by genome walking. Sequence analysis revealed two deduced open reading frames, pcaG and pcaH, encoding the α and β subunits of P34D respectively in this fragment. The P34D activity could be detected when pcaGH were expressed in E. Coli and the disruption of pcaH in strain NCIMB 10467 has lead to loss of its ability to catabolize protocatechuate. It was proved that the cloned pcaGH were encoding a functional protocatechuate 3, 4-dioxygenase which was necessary for the protocatechuate metabolism in this strain.

Objective To evaluate the effects of 131I treatment for Graves hyperthyroidism patients with leucopenia and the alteration of WBC levels after treatment.Methods A total of 257 Graves hyperthyroidism cases were retrospectively studied after 131I treatment.Based on baseline WBC count,119 cases with WBC count ＜4.0 × 109/L before 131I treatment were diagnosed with leucopenia and 138 cases had normal WBC.There were no significant differences in age,weight of thyroid,131I uptake rate in 24 h,dose of 131I and course of the disease between the two groups (all t ＜ 0.972,all P ＞ 0.05).WBC,lymphocyte,neutrophil and platelet counts were recorded before and 1,3,6 and 12 months after 131I therapy.The therapeutic effects were graded as clinical cure,improvement,invalidation and hypothyroidism.Statistical analyses,including independent samples t test,x2 test and one-way analysis of variance,were performed using SPSS 13.0.Results The WBC levels in the leucopenia group were (3.49 ± 0.43) × 109/L,(4.06 ±0.98) × 109/L,(4.20 ±1.04) × 109/L,(4.37 ±0.93) × 109/L and (4.88 ± 1.20) × 109/L before 131I treatment and 1,3,6,12 months after 131I treatment,respectively; while,those in the normal WBC group were (5.70 ± 1.08) × 109/L,(5.50 ± 1.14) × 109/L,(5.74 ±0.99) × 109/L,(5.95 ± 1.14) × 109/L and (6.07 ± 1.17) × 109/L,respectively.There was no statistically significant difference of WBC levels before and 1 month after 131 I treatment (t =1.662,P ＞ 0.05) in the normal WBC group,but WBC levels at those timepoints were significantly different in the leucopenia group (t =3.816,P ＜ 0.05).In the leucopenia group,there was no significant change of lymphocytes before and after 131I treatment,while the average neutrophil count showed an increasing trend during the 1,3,6 and 12 months after 131I treatment (F =40.583,t:1.468-11.264,all P ＜ 0.05).The average platelet counts at 6 and 12 months after 131I treatment were (187.80 ± 36.03) × 109/L and (206.88 ± 26.04) × 109/L respectively,which were higher than that before 131I treatment (F =9.735,t =2.604,4.892,all P ＜0.05).In the normal WBC group,there were no statistical differences of WBC changes before and after 131I treatment except for a lower lymphocyte count at 1 month after 131I treatment than that at baseline ((1.79 ± 0.37) × 109/L vs (1.99 ±0.63) × 109/L;F =12.868,t =3.284,both P ＜ 0.05).The treatment effects of the two groups were not significantly different (x2 =0.739,P ＞ 0.05).Conclusions 131I treatment presents similar therapeutic effects on Graves hyperthyroidism patients with leucopenia and those with normal WBC levels.WBC levels in patients with leucopenia may recover to baseline during the post-treatment follow-up.Thus 131I treatment is a safe and effective method for Graves hyperthyroidism patients with leucopenia.

BACKGROUND: Odontogenic infection factor has been given much importance in the study of the etiology of secondary trigeminal neuralgia,and the theory of jaw bone cavities is proposed. OBJECTIVE: To study the relationship of the jaw bone cavities and the etiology of trigeminal neuralgia.DESIGN: A self-controlled trial.SETTING: Department of oral and maxillofacial surgery of a university hospital.PARTICIPANTS: The patients with the trigeminal neuralgia were treated in Department of Oral and Maxillofacial Surgery, Affiliated Hospital of the Medical College of Qingdao University from February 1994 to December 2003, from whom 45 were selected for this study, including 15 males and 30 females with altogether 74 jaw bone cavities.METHODS: Curettage of the jaw bone cavities was performed in these cases, and visual analogue scale(VAS) was adopted for evaluation of the postoperative pain.MAIN OUTCOME MEASURES: ① VAS; ② Pathological examination and bacteria culture of the specimens.RESULTS: Pain relief was achieved in 33 cases(73.3% ) after the first surgery and in 10 cases(22.2% ) after a second or third surgery. In 2 cases (4.5%), the pain was alleviated but medication was still needed for pain control. Pathological examinations in most cases identified predominantly in flammatory and granulation tissues.CONCLUSION: Jaw bone cavities may be one of the major etiologic factors of trigeminal neuragia.

Objective:To detect the immune response in mice inoculated by DNA vaccine expressing chimeric gene gag gp120 of HIV 1.Methods:After BALB/C mice were immunized by the eukaryotic expression plasmid pVAXGE,the numbers of T lymphocyte subgroups of spleen,the killing rate of specific CTL from the immunized mice and the titers of serum antibodies during the different time were observed.Results:The lymphocytes of the immunized group by recombinant plasmid pVAXGE proliferated,the specific CTL killing rate of spleen was higher significantly than those of the control group( P

Objective To analyze the pregnancy outcomes and effect on fertility in women with Graves' disease after 131I therapy and the health status of their newborns.Methods From year 2009 to 2012,a total of 102 female patients with Graves' disease,who were pregnant during propylthiouracil (PTU)/L-thyroxine (euthyrox) maintenance treatment,were retrospectively recruited.The subjects were divided into 131I therapy group (n=57) and non-131I therapy group (n=45).The time interval between 131I (non-131I) therapy and getting pregnant were at least 6 months.They were followed for their pregnancy outcomes.During the pregnancy period,recorded parameters included serum thyroid biochemistry,dose of PTU/euthyrox,health conditions of the newborns and termination of pregnancy.Medication compliance,delivery mode,baby gender and body weight between the 2 groups were compared byx2 test,Mann-Whitney u test,Fisher exact test and two-sample t test.Results Ninety-seven patients had normal labor.In 131I therapy group and non-131I therapy group,15 vs 22 patients were treated by PTU,31 vs 18 by euthyrox and 11 vs 5 without any medication during pregnancy (x2=5.69,P＞0.05).The medical compliance (PTU and euthyrox: u=163.50,233.50,both P＞0.05) was similar between the 2 groups.The delivery mode (natural labor/cesarean: 36/19 vs 28/14,x2 =0.016),baby gender (male/female: 32/23 vs 18/24,x2 =2.239) and body weight ((3.18t0.47) vs (3.07±4.17) kg,t=1.154) were also similar between the 2 groups (all P＞0.05).Condusion Women of reproductive age with Graves' disease were found to have normal pregnancy after 131I therapy,on the condition that their thyroid function could be reasonably controlled and maintained by the proper medication before and during pregnancy.

Objective To investigate the inhibition effects of an hTERT-promoter-dependent oncolytic adenovirus Ad-VT that expresses apoptin on human prostatic carcinoma cell PC-3. Methods MTT assay was used to measure viability of PC-3 cell which was infected by recombinant adenovirus.The viability was measured at time points of 12,24,36,48,60,72,84 and 96 h after infection.AO/EB staining,DAPI staining,Annexin V assay were used to investigate the lethal effect and style of Ad-VT on PC-3 cell in vitro.The Caspases were measured by whole cell extraction of PC-3 cells 48hrs after infection. Results Ad-VT,Ad-VP3 and Ad-GT inhibited the proliferation of PC-3 cell in vitro.Ad-VT and Ad-GT were more effective than Ad-VP3 on cell growth,P ＜ 0.05.At 48,72,96 h time points,the inhibition effect of Ad-VT on PC-3 cell exhibited a dose related manner.When infection at MOI 100,the inhibition effect of Ad-VT on PC-3 cells exhibited time related manner.The AO/EB staining,DAPI staining,Annexin V assay,Annexin V assays and Caspase assays showed that Ad-VT inhibited the proliferation of PC-3 cells by inducing apoptosis of prostate cancer cells,Loss of cytoplasmic membrane integrity. Conclusions The hTERT-promoterdependent oncolytic adenovirus Ad-VT could effectively suppress prostate cancer cells PC-3 growth.

Objective:A new recombinant baculovirus transfer vector was constructed, in whic h a recombinant gene fragment encoding HIV-1 gag-gp120 chimeric gene was inse rted.Methods:After HIV-1 gag gene and gp120 gene were linked,the recombinant baculov irus vector was constructed,and the DNA recombinant technique and the E coli /baculovirus system were used.Results:Gel electrophoresis of DNA analysis showed that the genes were recombine d correctly.Electron microscope showed that the recombinant baculovirus reproduc ed in a great quantity.Conclusion:Recombinant baculovirus vector which HIV-1 gag-gp120 chimeric gene fragment was inserted in was constructed successfully.This vector is useful in study of the expression and the biological function of the HIV-1 Gag-gp120 ch imeric protein.

Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1)and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of infiuenza virus(H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus(H1N1)positiVe serum samples of mice.Results:Three Th/B cell epitopes containing HA_(73-87),HA_(125-139),HA_(188-205) were acquired Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus(H1N1)isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1)positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus(H1N1).Our study also establish the foundations for the further research of influenza virus infectlon and immunity mechanism,the recognition of influenza virus(H1N1)functional epitope and the development of epitope vaccines.

Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1) and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of influenza virus (H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus (H1N1) positive serum samples of mice.Results:Three Th/B cell epitopes containing HA73-87,HA125-139,HA188-205 were acquired.Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus (H1N1) isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1) positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus (H1N1).Our study also establish the foundations for the further research of influenza virus infection and immunity mechanism,the recognition of influenza virus (H1N1) functional epitope and the development of epitope vaccines.

BACKGROUND: Embryonic stem cells fESCs) develop from early blastular inner cell mass.Under proper condition,ESCs can maintain undifferentiated state in vitro,normal diploid nuclear type,and proliferative potential.In addition,ESCs possess multi-directional differentiation capacity and can differentiate into all sorts of cells of three germ layers. OBJECTIVE: To investigate the feasibility of directed differentiation of ESCs towards thyroid cells in vitro as well as related molecular expression. DESIGN,TIME AND SETTING: A cell-based observational experiment was performed at the Bank of Cord Blood,Second Hospital Affiliated to Sun Yat-sen University between January 2004 and December 2006.MATERIALS: Balb/c pregnant mice at pregnancy 12.5-14.5 days were used for preparation of embryonic fibroblast feeder layer.E14 mouse embryonic stem cell (ESC) strains were gifted by professor Xu from Harvard University.METHODS: Murine El4 ESCs were cultured in methylcellulose semisolid medium to form embryoid bodies (Ebs).These Ebs were transferred for further inductive culture with the stepwise addition of growth factors- thyrotropin (TSH),insulin and kalium iodidum (KI).The cultured thyroid cells of adult Kunming mice were taken as positive control.MAIN OUTCOME MEASURES: ①Cellular morphological change in the process of differentiation.② Detection of expression levels of TSHR,PAX8,TTF-2,and TIF-1 by immunofluorescence assay.③ Detection of expression levels of TSHR,PAX8,NIS,TPO,and Tg by reverse transcription-polymerase chain reaction (RT-PCR).RESULTS: The differentiated cells had clear boundary,exhibiting round,oval,shuttle-shaped,or polygon adhesive growth.On day 6 of inductive ifferentiation,the differentiated cells showed the expression of PAX8,NIS,TPO,Tg,and TSHR,the specific gene of thyroid ceils.On day 8 of inductive differentiation,the expression of TSHR,TIF-1,PAX8,and TIF-2 was detectable in the differentiated cells with morphous similar to thyroid cells.CONCLUSION: ESCs can differentiate towards thyroid cells under given inductive conditions.