Objective To observe the effect of Anshen Bunao syrup combined with escitalopram oxalate on nervous function defect in patients with post stroke depression,and to provide effective treatment for the disease with integrated traditional Chinese and western medicine.Methods 257 patients with post stroke depression were selected,and they were divided into observation group(130 cases) and control group(127 cases) according to the digital table.The two groups were given the same basic treatment,the control group was given escitalopram oxalate,and the observation group was given Anshen Bunao syrup combined with escitalopram oxalate.The NIHSS and HAMD scores of the two groups were evaluated before and 2,4,8 weeks after treatment.The adverse drug reaction was observed during the treatment period.The treatment effect was determined according to the HAMD reduction rate after 8 weeks.Results 2,4 and 8 weeks after treatment,the NIHSS scores of the observation group were (18.9 ±6.2)points,(15.6 ± 6.4) points,(12.2 ± 4.3) points,respectively,which were lower than (22.5 ± 7.1) points before treatment (t =4.355,8.230,14.148,all P ＜ 0.05).4 and 8 weeks after treatment,the NIHSS scores of the control group were (17.2 ± 7.5) points,(14.4 6.6) points,respectively,which were lower than (22.1 ± 8.4) points before treatment (t =4.904,8.123,all P ＜ 0.05).4 and 8 weeks after treatment,the NIHSS scores of the observation group were lower than those of the control group(t =3.640,1.855,all P ＜ 0.05).2,4 and 8 weeks after treatment,the HAMD scores of the two groups[(15.4 ± 5.1) points,(14.0 ± 5.2) points,(10.5 ± 4.7) points and (14.8 ± 4.6) points,(12.4 ± 5.8) points,(7.9 ± 3.8) points] were lower than before treatment [(23.9 ± 4.7) points,(23.2 ± 5.4) points] (t =13.501,15.539,26.419,13.812,4.748,4.748,all P ＜ 0.05).8 weeks after treatment,the HAMD score of the observation group was lower than that of the control group (t =3.640,P ＜ 0.05).8 weeks after treatment,the treatment effect was determined according to the HAMD reduction rate,the total effective rates of the observation group and the control group were 95.4％ (124/130),91.3 ％ (116/127),respectively.The curative effect of the observation group was better than the control group(Z =3.104,P ＜ 0.05).In the course of treatment,the incidence rate of adverse drug reactions of the control group was 6.30％ (8/127),which of the observation group was 5.38％ (7/130),there was no statistically significant difference between the two groups(x2 =0.098,P ＞ 0.05).Conclusion Anshen Bunao syrup combined with escitalopram oxalate can improve the nervous function defect in patients with post stroke depression,and the incidence rate of adverse drug reactions is low,which has good clinical value.

Objective To explore the value and applicability of the activity of glucose-6 phosphate dehydrogenase(G-6PD) for the auxiliary diagnosis of thalassemia.Methods Nine hundred and forty samples verified by the agar gel hemoglobin electrophoresis and(or) gene diagnosis,blood count measurement,serum ferritin and G-6PD activity test were divided into 3 groups [820 cases of thalassemia in group A;40 cases of iron deficiency anemia(IDA) in group B;80 cases normal control group in group C] and the G-6PD activities of them were analyzed statistically.Results The activity of G-6PD of those samples were(35.23?7.11),(34.95?10.72),(26.64?10.85),(23.86?7.68),(19.89?5.99),(18.65?6.67),(16.75?5.49) NBTU respectively in HbH disease,?-thalassemia major,?-thalassemia intermedia,IDA,?-thalassemia minor,? combine with ?-thalassemia,?-thalassemia trait,there were significant differences compared with normal control group(Pa0.05).Conclusions G-6PD activity increase in both thalassemia and IDA group,it can be used in auxiliary diagnosis of thalassemia but had its serviceable range.It′s suitable for the auxiliary diagnosis of HbH disease and ?-thalassemia major,but not for the discrimination of gene category in thalassemia.

Objective To construct, express and identify the anti-idiotypic antibody single chain variable fragment (scFv) against Edwardsiella tarda. Methods By using RT-PCR method, the variable regions of the heavy and light chain of the anti-idiotypic monoclonal antibody (mAb) 1E11 against Edwardsiella tarda were cloned and joined with a (Gly_4ser)_3 linker, and the scFv in the orientation of V_L-linker-V_H was constructed. It was then cloned into vector plasmid pET-28a, expressed in Escherichia coli BL21(DE3), and confirmed by SDS-PAGE, Western blot and ELISA. Results The recombinant scFv could be expressed in E.coli BL21 (DE3) in a fusion protein pattern. The expression product was in the form of an inclusion body and the purified fusion protein was obtained after being purified and refolded. The SDS-PAGE and Western blot analysis showed that the molecular had the binding activity to the antigen. Conclusion The recombinant anti-idiotype scFv has been successfully constructed and expressed in E.coli BL21 (DE3), providing the basis and potential for preparation of genetically engineered vaccine against Edwardsiella tarda.

BACKGROUND: Cytokine is a poly-functional and effective regulator factor to regulate growth of multiple cells. Researches suggest that, as an inflammatory medium, cytokines play a key role in inflammatory reaction of eye; however, there are rare studies on dynamic changes after traumatic cataract.OBJECTIVE: To investigate the relationship among inflammatory reaction and dynamic changes of interleukin 1 (IL-1),IL-6 and tumor necrosis factor alpha (TNF-α) in aqueous humor of rabbits after extracapsular cataract extraction.DESIGN: Randomized controlled animal study.SETTING: Shenzhen Municipal Ophthalmology Hospital, Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University.MATERIALS: The experiment was carried out in Institute of Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University from March 2004 to March 2006. Fifteen healthy adult New Zealand rabbits of 30 eyes,general grade, weighting 2.5-3.0 kg, of either gender, were provided by Animal Center of the Second Clinical Medical College of Jinan University [certification: SYXK (yue) 2005-006]. Eyes of rabbits were normal before experiment. All rabbits were randomly divided into normal control group, traumatic control group and operative group with 5 in each group and in total of 10 eyes in each group. IL-1, IL-6 and TNF-α kits were provided by Shenzhen Yawei Biotechnology Company Limited.METHODS: Rabbits in traumatic control group and operative group were totally anesthetized with intravenous injection of 10% 1 mi/kg urethan, and then, 5# needle was punctured from corneal limbus to anterior chamber to scarify anterior membrane of lens about 5 mm to establish animal models of traumatic cataract of oculus uterque. Rabbits in normal control group were fed normally. After successful modeling, common antibiotic eyedrops was used to clean conjunctival sac of rabbits in traumatic control group and operative group 3 times a day. On the 3rd day of successful modeling, rabbits in operative group were totally anesthetized with intravenous injection of 10% 1 mL/kg urethan, and then, they undertook extracapsular cataract extraction of oculus uterque: horizontally intercepting bladder or waterly separating with breakage of anterior bladder membrane, expulsing nucleus of lens, washing lens cortex and suturing incisions. On the operative day and on the 1st, 3rd, 7th and 14th days after operation, inflammatory reaction of anterior chamber in traumatic control group and operative group was measured and 0.2 mL aqueous humor was extracted from rabbits in three groups to count and classify cells; meanwhile, expressed level and dynamic changes of IL-1, IL-6 and TNF-α in cytokines of aqueous humor were measured with double-antibodies ELASA technique.MAIN OUTCOME MEASURES: Total numbors of leucocytes and contents of IL-1, IL-6 and TNF-α in aqueous humor of rabbits in three groups after operation.RESULTS: ① On the 1st, 3rd, 7th and 14th days after operation, numbers of leucocytes were (2.4±0.7)×106/L, (2,2±0.5)×106/L, (2.8±0.8)×106/L and (2.0±0.5)×106/L in aqueous humor; (19.7±7.3)×106/L, (28.1±9.6)×106/L, (14.2±5.6)×106/L and(8.4±3.8)×106/L in traumatic control group; (65.3±14.5)×106/L, (79.8±12.7)×106/L, (21.7±8.2)×106/L and (12.4±4.1)×106/L in operative group. In addition, numbers of leucocytes were more in traumatic control group and operative group than those in normal control group (F =22.5, 27.9, 11.6, 8.4;P＜0.05). ② Within 1-14 days after operation, contents of IL-1, IL-6and TNF-α in aqueous humo were higher in traumatic control group and operative group than those in normal control group (P＜0.05), and there was a significant difference between traumatic control group and operative group (P＜0.05);however, there was no significant difference among three groups on the operative day (P＞0.05). ③ Contents of cytokines reached peak on the 7th day after operation, decreased gradually, and reached the lowest value on the 14th day.Contents in traumatic control group and operative group were higher than those in normal control group (P＜0.05).CONCLUSION: The intraocular inflammation after lens extraction is closely related to the dynamic changes of IL-1, IL-6and TNF-α levels in aqueous humor. Cytokine may be one of crucially inflammatory agents in the eyes after traumatic cataract.

Objective To explore the value of brainstem auditory evoked potential(BAEP),visual evoked potential(VEP)and Revised Visual Retention Test(VRT)in detecting impairment of central nervous system in patients with acute leukemia(AL).Methods Neuropack-Ml type evoked potential machine(Japan)was used to check brainstem auditory evoked potential(BAEP)and visual evoked potential(VEP).BAEP,VEP and VRT were examined in 30 cases with acute leukemia(group AL)and in 30 age and sex matched healthy subjects as a control(groupC).Indexes of BAEP including peak latency(PL)of waves I ,IB ,V and inter-peak latency(IPL)of waves Ⅰ～Ⅲ,Ⅲ ～Ⅴ,Ⅰ～Ⅴand indexes of VEP(P100PL)were determined.VRT was accomplished meanwhile applying B method with C pattern,C method with D pattern,D method with E pattern.The correct scores and error scores were recorded separately.Results BAEP:the abnormal rate of the group AL was 23.3 %.Compared with control group,we found significant longer latency of wave Ⅲ、Ⅴ(P＜0.05)and significant elongation in the mean inter-peak latencies of Ⅰ～Ⅴ(P＜0.05).VEP:the abnormal rate of the group AL was 26.7% ,the P100 PL of the patients was longer than that in the control group(P＜0.05).VRT:the abnormal rate of the group AL was 33.3%.The results of VRT showed that the score in the group AL in D method with E pattern was lower than that in the control group(P＜0.05).Conclusion BEAP,VEP and VRT could detect the impairment of central nervous system in patients with acute leukemia sensitively,and give clues to early diagnosis of central nervous system leukemia(CNSL).

Objective To study inflammatory cytokines changes in brain tissue of neonatal rats with periventricular leukomalacia (PVL).Method A total of 80 neonatal SD rats (P3) were randomly assigned into 2 equal groups,sham-operated group and PVL group.Rats in each group were further assigned into four subgroups (12,24,48,72 h),with 10 rats in each subgroup.The hypoxic-ischemic PVL modal were established following the procedure:first,isolation and ligation of left common carotid artery,and then exposed to 8％ O2 and 92％ N2 for 2.5 h.The sham-operated rats were processed with isolation of left common carotid artery only.Rats of the four subgroups were sacrificed at 12,24,48 h and 72 h respectively,then the brains were rapidly removed in corresponding time.Pathological changes of brain tissues were observed using HE stain.The mRNA expression levels of tumor necrosis factor α(TNF-α)and inter leukin-1β (IL-1β) were assessed using real-time quantitative PCR assays,the protein levels of TNF-α and IL-1β were detected using enzyme linked immunosorbent assay method.Result The brains tissues of rats in PVL group showed remarkably hyperemia and edema,with left ventricle enlargement.Periventricular white matter structure was disintegrated comparing with sham-operated group.The expression of TNF-α and IL-1 β mRNA in PVL group increased significantly,reaching peak by 24 h and then gradually decreased 72 h after the procedure.The mRNA levels of TNF-c and IL-1 β were significantly different between each two time points of 12,24 h and 48 h in PVL group (P ＜0.05).However,there was no differences between 72 h and 48 h within PVL and sham-operated group group(P ＞0.05).In PVL group,the protein expression trends of TNF-α and IL-1 β were similar to mRNA expression trends.Moreover,the protein levels were significantly different between each two time points of TNF-α and IL-1 β,respectively (P ＜ 0.01).The protein expression levels of TNF-α were different at each time point between PVL group and sham-operated group[(189.2 ± 20.4) pg/ml vs.(131.4 ±5.2) pg/ml at 12 h,(213.8 ± 16.7) pg/ml vs.(127.7 ±7.4) pg/ml at 24 h,(181.7 ± 15.0) pg/ml vs.(126.3 ± 6.0) pg/ml at 48 h,(159.6 ± 25.3) pg/ml vs.(131.4 ± 6.0) pg/ml at 72 h;P ＜0.01].The protein levels of IL-1β were different between the two groups only at 24 h and 48 h.[(121.8 ±30.0) pg/ml vs.(67.4 ± 13.7) pg/ml,(83.3 ± 15.7) pg/ml vs.(65.3 ± 14.9) pg/ml;P ＜0.05].In sham-operated group,no differences of TNF-α and IL-1 β protein levels were found between any different time points (P ＞ 0.05).Conclusion Inflammatory cytokines such as TNF-α and IL-1β are involved in ischemic-hypoxia induced PVL.Dynamic detection of inflammatory factors is expected to be an important method of early diagnosis,assessment of treatment efficacy and prognosis of PVL.

Objective To evaluate the cytogenetic, molecular responses and safety of generic imatinib in newly diagnosed patients with chronic myelogenous leukemia in chronic phase (CML-CP) in 1-year at different stages. Methods From January 2014 to November 2014, 50 CML-CP patients received oral generic imatinib 400 mg/d. The cytogenetic examinations, bcr-abl transcript levels and safety were monitored after 3, 6, 9 and 12 months respectively. Results 46 of 50 patients insisted on oral generic imatinib and followed up 1 year. At 3-month, 52.0 % (26/50) patients reached the complete hematologic responses (CHR) rate, and patients at least achieved minor cytogenetic response (mCyR) and bcr-ablIS≤10 % were 84.0 % (42/50) and 42.0 % (21/50). At 6-month, patients at least achieved part cytogenetic response (PCyR) and bcr-ablIS≤10 %were 73.5 % (36/49) and 59.2 % (29/49). At 12-month, patients achieved complete cytogenetic response (CCyR), bcr-ablIS≤1 % and bcr-ablIS≤0.1 % were 60.9 % (28/46), 63.1 % (29/46) and 45.7 % (21/46). The grade 3 leukopenia, thrombocytopenia and anemia rates were 34 % (17/50), 40 % (20/50) and 30 % (15/50), respectively. No grade 4 hematologic toxicity occurred. The common non-hematologic toxicities included edema [84 % (42/50)], nausea [46 % (40/50)], muscle pain [20 % (10/50)], rash [16 % (8/50)], and impaired liver function [8 % (4/50)]. Conclusion Generic imatinib has a favorable effect in treatment of patients with CML-CP, and without serious adverse reactions.

Objective To construct,express and identify the anti-idiotypic antibody single chain variable fragment(scFv) against Edwardsiella tarda.Methods By using RT-PCR method,the variable regions of the heavy and light chain of the anti-idiotypic monoclonal antibody(mAb) 1E11 against Edwardsiella tarda were cloned and joined with a(Gly4Ser)3 linker,and the scFv in the orientation of VL-linker-VH was constructed.It was then cloned into vector plasmid pET-28a,expressed in Escherichia coli BL21(DE3),and confirmed by SDS-PAGE,Western blot and ELISA.Results The recombinant scFv could be expressed in E.coli BL21(DE3) in a fusion protein pattern.The expression product was in the form of an inclusion body and the purified fusion protein was obtained after being purified and refolded.The SDS-PAGE and Western blot analysis showed that the molecular weight of scFv protein was 27 ku.Indirect ELISA confirmed that the scFv had the binding activity to the antigen.Conclusion The recombinant anti-idiotype scFv has been successfully constructed and expressed in E.coli BL21(DE3),providing the basis and potential for preparation of genetically engineered vaccine against Edwardsiella tarda.

Objective To detect and analyse acquired pure red cell aplasia (PRCA) T lymphocyte subsets distribution and to assess its condition and cyclosporine immune function of T lymphocytes subsets.Methods Flow cytometry was applicated to detect peripheral blood T lymphocyte subsets of acquired PRCA patients before and after 3 months of treatment with cyclosporine-based immunosuppressive regimen and normal controls.Results Among 22 patients,17 cases of blood returning normal (three cases of bone marrow returning normal),the total effective rate was 95.5 ％ (3 cases of cure,14 cases of remission,4 cases of improvement,1 case of ineffectiveness).Th (CD3+ CD4+) cells and Th/Ts ratios in acquired PRCA patient group were lower than those in the normal control group,while Ts (CD3+ CD8+) cells was higher than that in the normal control group,the differences were statistically significant (P ＜ 0.05).After 3 months of treatment,Th cells and Th/Ts ratio were higher than those before,and Ts cells were decreased compared with the previous (P ＜ 0.05).Conclusion Disorders of T lymphocyte subsets in acquired PRCA patients lead to immune dysfunction,however,cyclosporine can improve T lymphocyte subsets in patients with imbalance,which is an effective way to treat the disease with significant curative effect and mild adverse reactions.

Objective To investigate the clinical features of rheumatoid arthritis (RA) associated with thyroid function abnormalities.Methods Serum thyroid function,anti-thyroglobulin antibodies (TGAb) and anti-thyroid peroxidase antibodies (TPOAb) were detected in 186 RA patients.Clinical data were then analyzed.Comparison of continuous data between groups was made by t test,while comparison of categorical data was made by x2 test.The non-normal distribution of the data was analyzed by Mann-Whitney U rank sum test.Results Fifty-six RA cases (30％) had abnormal thyroid function,including 25 low T3 syndrome cases,16 subclinical hypothyroidism cases,10 hypothyroidism cases and 5 Graves' disease cases.Among them 23 cases were diagnosed as Hashimoto's thyroiditis.The positive rate in the hypothyroidism group with chills (45 cases,80.3％,x2=99.94),lazy words (14 cases,25.0％;x2=7.896),lethargy (24 cases,42.9％;x2=7.433) and abdominal distension (21 cases,37.5％;x2=7.15) was higher than that in the group with normal thyroid function (all P＜0.05).Level of serum high density lipoprotein was decreased in hypothyroid group [(1.0±0.5) mmol/L vs (1.4±0.6) mmol/L,t=4.087,P＜0.05].Patients with abnormal thyroid function had higher positive rate of ANA [(10 cases,7.7％) vs (25 cases,44.5％),x2=34.98],anti-SSA [(3 cases,2.3％) vs (23 cases,41.1％),x2=48.91],anti-SSB [(2 cases,1.5％) vs (12 cases,21.4％),x2=22.25],TGAb [(20 cases,15.4％) vs (43 cases,76.7％),x2=65.88] and TPOAb [(13 cases,10.0％) vs (27 cases,48.2％),x2=35.90] than those with normal thyroid function (all P＜O.05).Conclusion Rheumatoid arthritis is often accompanied by thyroid disease.The positive rate of auto-antibodies in RA with abnormal thyroid function is higher than those with normal thyroid function and has no relationship with the disease activity.