The build-in low-pressure monolithic column combined with hydride generation atomic fluorescence spectrometry ( HG-AFS ) was employed for speciation analysis of fish meat. The sample pretreatment and separation approach could be accomplished within 30 min. The proper amount of fish sample was weighed and smashed into puree. The extraction solution composed of 10% HCl, 1% thiourea, and 0. 15% KCl was added before loaded into the automatic temperature controlled vertex system with 2000 r/min. The sample solution was separated through Merck monolithic column, with 3% ( V/V) acetonitrile, 30 mmol/L amonium acetate and 0. 03%(V/V) 2-mercaptoethanol (2-ME) as the eluent. The after-column eluent was digested by novel UV digestion device with pipeline sintered into the lamp, and then detected by hydrid-generation AFS. The rapid LC separation enabled fast mercury speciation of fish sample within 10 min. The different UV lamp digestion effects, eluent components, carrier gas, shield gas, lamp current, as well as PMT working power was optimized. Under the optimal conditions, the robust system achieved detection limits (DL) of 0. 15 μg/L and 0. 14 μg/L for methylmercury and HgⅡ, respectively. The RSD (n=7) was less than 5%, the linear correlation coefficient was 0 . 999 , and the matrix spiked recovery was in the range of 85%-110% for Hg speciation. This method was used for the determination of Hg speciation in fish and soil samples, and was proofed to be a reliable, easy approach for daily inspection.

Objective To explore the imaging differences of vertebral multiple myeloma(VMM)and vertebral osteolytic metastasis on CT. Methods Review the imaging signs of 32 patients with VMM and 52 patients with vertebral osteolytic metastasis,then record and finally statistical analysis was carried out.Results Compare to 1 67 vertebras involved in 52 cases of vertebral metastasis,the patents with VMM had 220 vertebras involved.The incidence of the multiple small circular type of bone destruction in VMM was 42.66%(93/218),which was higher than that in vertebral metastasis 0.00% (0/165)(χ2 =92.963,P =0.000).The incidence of the irregular shape type of bone destruction in VMM was 23.39% (5 1/218),while it was higher in patients with vertebral metastasis 45.45%(75/165)(χ2 =20.704,P=0.000).It was also found that the incidence of the fragmentary type of bone destruction and the involvement of the unilateral pedicle in VMM were lower than that in patients with vertebral metastasis 8.7% (19/218 )& 27.27% (45/165)(χ2 =23.238,P =0.000), 6.82% (1 5/220)& 1 7.96% (30/1 67)(χ2 =1 1.477,P =0.001).The incidence of the crest protrusion type of bone destruction in patients with VMM was 16.06% (35/218),however it was 9.10% (15/165)(χ2 =4.013,P =0.045)in vertebral osteolytic metastasis. Conclusion The imaging features of VMM and vertebral osteolytic metastasis had certain characteristic.And they can be differentiated from each other,combining with clinical traits.

Objective To monitor the expression patterns of bcr-abl in chronic myeloid leukemia (CML) patients during treatment with imatinib mesylate and evaluate the detection of MRD by RQ-PCR method. Methods The ABI Prism 7500 Sequence Detection System using Taqman fluorogenic probes was used to quantify target gene. bcr-abl mRNA was detected by RQ-PCR in 106 CML patients. The normalized quotient (NQ) of bcr-abl mRNA was calculated as followings: NQ=bcr-abl mRNA copy numbers/abl mRNA copy numbers. Results The NQ of BCR-ABL mRNA was well correlated with the progression of disease and the number of Ph+ cell (r =0.9824 and 0.9346, respectively). The NQ was decreased rapidly in 62 patients and kept in low level for a long time, and only 2 of them were relapsed. For 8 patients, after treatment the NQwere decreased initially and increased sharply, 7 of them were relapsed after 5-9 months. After treatment the NQ of 31 patients were still>0.1, 11 patients were relapsed after a short remission and 7 were ineffective or progression. Out of 5 patients whose NQ were fluctuated and had little regularity, but all of them had a continuing remission. Conclusion RQ-PCR is a more sensitive technique in the detection of bcr-abl fusion gene.It is an important method to monitor the tumor cell during the treatment with imatinib mesylate in CML patients.

Objective: To evaluate the relationship between serum mRNA level of heparin binding epidermal growth factor (HB-EGF) and acute coronary syndrome (ACS) occurrence. Methods: Our research included in 2 groups: ACS group,n=50 patients and Control group,n=100 normal subjects. Serum HB-EGF mRNA level was examined by RT-PCR and the relationship between HB-EGF mRNA and ACS occurrence was assessed by Logistic regression analysis. Results: Compared with Control group, the serum HB-EGF mRNA level of ACS group was higher (0.22±0.73) vs (0.46±0.14),P<0.05. With adjusted meaningful factors of hypertension, smoking, TG, TC, LDL-C, HDL-C and BMI by single factor analysis, multi Logistic regression analysis indicated that serum HB-EGF mRNA was related to ACS occurrence (OR=5.813, 95% CI 2.342-14.426,P<0.001) which meant that upon 0.1 grey value of HB-EGF mRNA elevation, the risk of ACS occurrence may increase 4.813 folds accordingly. Conclusion: Serum HB-EGF mRNA level was related to ACS occurrence.

Objective To explore the effect of arsenic trioxide (As2O3) on the migration of neurons and the potential mechanism through the establishment of primary neuron culture from the brains of neonatal rats.Methods Brain tissues were selected from SD neonatal rats for primary neuron calture.The cells were divided into 4 groups based on the addition of As2 O3:normal control group,1 μmol/L As2O3 group,10 μmol/L As2O3 group and 20 μmol/L As2O3 group.The primary neurons were treated with different concentrations of As2O3 and cultured for 24 hours.Boyden chamber assay was used to detect the effect of As2O3 on neuronal migration.Immunofluorescence laser confocal microscope was used to observe the structure of actin.Results In the control group,the cultured neurons showed a regular pattern of distribution.In the 3 groups treated with As2O3,the distribution of neurons was loose and disordered,which was most obvious in the 20 μmol/L As2O3 group.The results showed that the higher concentration of As2O3,more difficult it was for the neurons to survive.The number of neuronal migration was 64.6 ± 4.3 for normal control group,63.0 ± 7.0 for 1 μmol/L As2O3 group,54.8 ± 3.6 for 10 μmol/L As2O3 group,and 21.6 ± 3.9 for 20 μmol/L As2O3 group.The results showed that As2O3 might inhibit the migration of primary neurons in a dose-dependent manner (F =49.31,P ＜0.001).The normal actin skeleton was destroyed under the laser confocal microscope in 10 μmol/L As2O3 group and 20 μmol/L As2O3 group,while they remained unaffected in normal control group and 1 μmol/L As2O2 group.Conclusion As2 O3 exposure can reduce the neuron migration in a dose-independent manner probably through disrupting the organization of acting cytoskeleton.

Objective This study was to investigate the computed tomography(CT)features differentiating tuberculous spondyli-tis from pyogenic spondylitis.Methods The CT features in 32 patients with tuberculous spondylitis and 30 patients with pyogenic spondylitis were retrospectively reviewed,and statistically analyzed.Results In 32 cases of tuberculous spondylitis,71 vertebra were involved.In 30 cases of pyogenic spondylitis,59 vertebra were involved.The incidence of thoracic vertebra involvement in tuberculous spondylitis was 60.56% (43/71),which was higher than that in pyogenic spondylitis (25.42%,1 5/59)(P <0.05).The incidence of lumbar vertebra involvement in tuberculous spondylitis was 33.80% (24/71 ),which was lower than that in pyogenic spondylitis (61.02%,36/59)(P <0.05).The incidence of the worm-eaten type of bone destruction in tuberculous spondylitis was 9.90% (7/71),which was lower than that in pyogenic spondylitis (44.07%,26/59)(P <0.05).The incidence of the fragmentary type of bone destruction in tuberculous spondylitis was 1 9.72%(14/71 ),which was higher than that of in pyogenic spondylitis (3.39%,2/59) (P <0.05).The incidence of large osteosclerosis in tuberculous spondylitis was 52.1 1%(37/71),which was higher than that in pyo-genic sp-ondylitis (22.03%,13/59),(P <0.05).The incidence of the involved vertebral height on sagital CT scan (less than the 1/2 of the normal vertebral height)in tuberculous spondylitis was 1 6.9% (12/71),which was lower than that in pyogenic spondylits (62.71%,37/59)(P <0.05).The incidence of patchy high density shadow in tuberculous spondylitis was 50.7% (36/71 ),which was higher than that in pyoge-nic spondylitis (20.34%,12/59)(P <0.05.)The incidence of the involvement of the appendages in tu-berculous spondylitis was 25.35% (18/71),which was higher than that in pyogenic spondylitis (8.47%,5/59)(P <0.05).The inci-dence of paravertebral abnormal soft tissue with calcification in tuberculous spondylitis was 60.00%(18/30),which was higher than that in pyogenic spondylitis(20.00%,5/25 )(P < 0.05 ).Conclusion Tuberculous spondylitis and pyogenic spondylitis have some characteristic imaging features,combined with the clinical signs differentiation diagnosis can be made each other.

[Objective] : To explore the clinical characteristics and methods for syndrome differentiation prediction, as well as to construct a predictive model for Qi deficiency and blood stasis syndrome in patients with acute ischemic stroke (AIS). [Methods] : This study employed a retrospective case-control design to analyze patients with AIS who received inpatient treatment at the Neurology Department of The First Hospital of Hunan University of Chinese Medicine from January 1, 2013 to December 31, 2022. AIS patients meeting the diagnostic criteria for Qi deficiency and blood stasis syndrome were stratified into case group, while those without Qi deficiency and blood stasis syndrome were stratified into control group. The demographic characteristics (age and gender), clinical parameters [time from onset to admission, National Institutes of Health Stroke Scale (NIHSS) score, and blood pressure], past medical history, traditional Chinese medicine (TCM) diagnostic characteristics (tongue and pulse), neurological symptoms and signs, imaging findings [magnetic resonance imaging-diffusion weighted imaging (MRI-DWI)], and biochemical indicators of the two groups were collected and compared. The indicators with statistical difference (P < 0.05) in univariate analysis were included in multivariate logistic regression analysis to evaluate their predictive value for the diagnosis of Qi deficiency and blood stasis syndrome, and the predictive model was constructed by receiver operating characteristic (ROC) curve analysis. [Results] : The study included 1 035 AIS patients, with 404 cases in case group and 631 cases in control group. Compared with control group, patients in case group were significantly older, had extended onset-to-admission time, lower diastolic blood pressure, and lower NIHSS scores (P < 0.05). Case group showed lower incidence of hypertension history (P < 0.05). Regarding tongue and pulse characteristics, pale and dark tongue colors, white tongue coating, fine pulse, astringent pulse, and sinking pulse were more common in case group. Imaging examinations demonstrated higher proportions of centrum semiovale infarction, cerebral atrophy, and vertebral artery stenosis in case group (P < 0.05). Among biochemical indicators, case group showed higher proportions of elevated fasting blood glucose and glycated hemoglobin (HbA1c), while lower proportions of elevated white blood cell count, reduced hemoglobin, and reduced high-density lipoprotein cholesterol (HDL-C) (P < 0.05). Multivariate logistic regression analysis identified significant predictors for Qi deficiency and blood stasis syndrome including: fine pulse [odds ratio (OR) = 4.38], astringent pulse (OR = 3.67), superficial sensory abnormalities (OR = 1.86), centrum semiovale infarction (OR = 1.57), cerebral atrophy (OR = 1.55), vertebral artery stenosis (OR = 1.62), and elevated HbA1c (OR = 3.52). The ROC curve analysis of the comprehensive prediction model yielded an area under the curve (AUC) of 0.878 [95% confidence interval (CI) = 0.855 – 0.900]. [Conclusion] This study finds out that Qi deficiency and blood stasis syndrome represents one of the primary types of AIS. Fine pulse, astringent pulse, superficial sensory abnormalities, centrum semiovale infarction, cerebral atrophy, vertebral artery stenosis, elevated blood glucose, elevated HbA1c, pale and dark tongue colors, and white tongue coating are key objective diagnostic indicators for the syndrome differentiation of AIS with Qi deficiency and blood stasis syndrome. Based on these indicators, a syndrome differentiation prediction model has been developed, offering a more objective basis for clinical diagnosis, and help to rapidly identify this syndrome in clinical practice and reduce misdiagnosis and missed diagnosis.

To identify SJCHGC01743 gene of Schistosoma japonicum and evaluate the potential of the recombinant protein as a new vaccine candidate for schistosomiasis, polymerase chain reaction (PCR) technique was used to amplify the cDNA of the gene and real-time RT-PCR was used to analyze the transcription profiles of SJCHGC01743 at different development stages. Recombinant plasmid was successfully constructed and transformed into competent Escherichia coli BL21 (DE3). Then the recombinant protein was expressed, purified and emulsified with ISA206 adjuvant to immunize BALB/c mice for three times. The immunogenicity was confirmed by Western blotting and tissue localization was detected by indirect immunofluorescent assay. The specific antibody level was detected by ELISA. The immunoprotection of rSjOST48 was evaluated by the reduction in worm and egg counts in mice. A cDNA with 1 248 nucleotides was isolated from 28-day-old schistosomes cDNAs by PCR. Sequence analysis revealed that SJCHGC01743 was a 48-kDa subunit of the oligosaccharyltransferase complex (OST48) and named as SjOST48. Real-time PCR analysis indicated that this gene was expressed in all investigated stages and had the highest expression level in 28 d worms, the level of gene transcription in female worms was significantly higher than that of male worms. Then recombinant plasmid pET28a(+)-SjOST48 was successfully constructed and expressed in E. coli BL21 (DE3). Western blotting analysis showed that rSjOST48 had good immunogenicity. Indirect immunofluorescent analysis revealed that SjOST48 was mainly distributed on the tegument of the worms. The result of ELISA indicated that the rSjOST48 vaccinated group could induce a significant increase in the level of specific IgG, IgG1 and IgG2a. An immunoprotection experiment showed that the vaccination of rSjOST48 in mice induced 32.62% (P < 0.05) reduction in the numbers of worms and 57.61% (P < 0.01) in eggs in liver, compared with that of the control group. This study provides the foundation for proceeding further research on the biological function of SjOST48 and screening new vaccine candidates for schistosomiasis.
Animals ; Antibodies, Helminth ; blood ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Female ; Genes, Helminth ; Helminth Proteins ; genetics ; immunology ; Immunoglobulin G ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Schistosoma japonicum ; genetics ; Schistosomiasis japonica ; prevention & control ; Vaccination

Objective:To explore the characteristics of immunological reconstitution of T-cell subsets and the role of Cornus,a Chinese medicinal herb on T lymphocytes in mice after irradiation (IR). Methods:Irradiated model mice were exposed to a single dose of X-ray radiation (2. 6 Gy) with or without Cornus treatments. Bloodroutine was examined before or after irradiation. CD3+,CD4+, CD8+ T cells and Th1, Tc1, Th2, Th17, Treg from spleen or peripheral blood were determined by flow cytometry. Results: Total lymphocytes and CD3+T cells including CD4+T and CD8+T cells,were significantly reduced 3 days after irradiation (P<0. 05). The re-constitution of CD3+ T cells ( especially CD8+T cells ) started from 5 days post irradiation, CD4+T cells increased 8 days after irradiation. However,the production of IFN-γ by Th1 or Tc1 cells were evidently decreased compared with control group even 8 days post irradiation (P<0. 05). In contrast to Th1,Th2/Th17/Treg were observably increased in irradiation group (P<0. 05). But the percentage of Th1 was obviously increased, and Th2, Th17, Treg were markedly decreased in cornus treated mice compared with irradiated mice (P<0. 05). Conclusion: The immunological reconstitution of T lymphocytes started from 5 days after total irradiation with rapid recovery of CD8+ T cells but not CD4+T cells. Cornus effectively improved the imbalance of T cell subsets by promoting the proliferation of Th1 and suppressing Th17 and Treg.

Objective To clone cDNA encoding troponin T of Schistosoma japonicum(SjTnT),and evaluate the protective efficacy induced by recombinant SjTnT in BALB/c mice against S. japonicum challenge infection. Methods The SjTnT gene was amplified from 28-day-schistosome cDNAs by PCR and then subcloned into pET28a(+). The recombinant SjTnT protein (rSjTnT)was expressed in Escherichia coli BL21(DE3)cells. The serum specific to rSjTnT was prepared by immunized BALB/c mice with the recombinant antigen,and the immunogenicity of rSjTnT was detected by Western blotting and ELISA. The immuno-protective efficacy induced by rSjTnT in BALB/c mice was evaluated according to the reduction in worm and egg counts. Results The cDNA encoding SjTnT was successfully cloned and expressed in E. coli. Western blotting showed that rSjTnT had a good immunogenicity. The high level of specific IgG antibodies was detected,and 33.89% worm reduction and 43.94% liver egg reduction were obtained in mice vaccinated with rSjTnT combined with Seppic 206 adjuvant compared with those in the adjuvant control group. Conclusions rSjTnT could induce partial immuno-protection against S. japonicum infec-tion in BALB/c mice. This study provided a basic for understanding the biological function of SjTnT.