The incidence of α-thalassemia and β-thalassemia is high in Guangxi,Guangdong,Sichuan and other province in China.Because no effective approach to thalassemia treatment could be used clinically now,the most cost-effective strategy to control this disease is to prevent the birth of babies with severe form of thalassemia.It is important to make effective screening and correct diagnosis of thalassemia by laboratory test.Laboratory diagnosis of thalassemia includes routine diagnosis and genetic diagnosis.The laboratory routine tests are some hematology examination,comprising red blood cell indices,erythrocyte osmotic fragility test,hemoglobin analysis,and others.Anyone alone of these laboratory parameters can not be used to diagnose the carrier of thalassemia.It is necessary to combine these tests to make screening diagnosis.The final diagnosis of thalassemia need to perform the gene mutation examination or globin train analysis.Technologies for gene mutation detection have been the main and gold standand method of diagnosing thalassemia now.

Objective In this paper,major issues for all those who have been selected in China national treatment program for patients with chronic Keshan disease (CNTP-CKD) were uncovered through evaluation of the annually reported data from participating provinces,in order to improve the performance quality of the program.Methods The datasets 2005-2012 were merged after cleaning them,and the composition of the treated patients was statistically analyzed,including gender and age distribution,diagnosis evidence for chronic Keshan disease (ECG,cardiothoracic ratio by X-ray,heart function grade of NYHA),and proportion of cases who had received treatment more than once.Results ①A total of 2 649 patients participated in the treatment,of them 1 115 patients were males accounting for 42.1％ (1 115/2 649),1 534 patients were females accounting for 57.9％ (1 534/2 649).Age of the patients were mainly distributed in 41 to 70 years old,and 24 CKD patients under 10 years accounting for 0.9％ (24/2 649).②2 313 cases of the involved patients were diagnosed with sufficient evidence,accounting for 81.9％ (2 313/2 823) and 121 cases with full misdiagnosis,accounting for 18.1％ (121/2 823).③There were 881 patients been treated for more than once,accounting for 38.3％ (881/2 301) of the number of treatment.Conclusions ① Diagnosis for CKD remains a key problem,suggesting that medical record for each patient diagnosed by province-level doctors' needs to be built up as early as possible.The rate of patient treatment for more than once is low which is not beneficial to the patients.② Treatment period for CKD patients is highly recommended to expand to at least one year,and the disease should be enrolled in the free cost list of the new rural cooperative medical system (NCMS).

Objective:To explore the role CD4+CD45RO+memory T cells in the pathogenesis of Hashimoto's thyroiditis (HT) by detecting the percentages of CD4+CD45RO+ memory T cells in peripheral blood mononuclear cells in peripheral blood of newly diagnosed HT patients. Methods:53HT patients and 43 matched healthy controls (HC) were included in this study. According to the thyroid functions,HT patients were divided into euthyroid subset(HT-A,n =15) ,subclinical hypothyroidism(HT-B,n=14) and overt hypothyroidism subset (HT-C,n=24). The percentages of CD4+CD45RO+memory T cells in PBMCs,as well as the level of serum IFN-γ and IL-17,and thyroid functions,and the titers of thyroid-specific autoantibodies (TPOAb,TgAb) were respectively detected by flow cytometry,ELISA,and ECLIA. Results:The percentages of CD4+CD45RO+ memory T cells in PBMCs,as well as the level of serum IFN-γ and IL-17,the titers of TPOAb,TgAb were all significantly higher than that in HC(P<0. 01). Bivariate correlation revealed that the percentages of CD4+CD45RO+ memory T cells positively correlated with the level of serum IFN-γ,TPOAb and TgAb(P<0. 01,P=0. 015,P<0. 01) in HT patients. Conclusion:The significant increase of CD4+CD45RO+memory T cells in peripheral blood of patients with HT suggested a role of CD4+CD45RO+ memory T cells in the pathogenesis of this disease.

Objective:To study the effect of microRNA (miR)-330-3p on hepatic ischemia-reperfusion injury (IRI) in mice, meanwhile, and to determine potential molecular mechanism.Methods:Eighty male C57BL/6 mice, aged 7-8 weeks, 23-25 g, specific pathogen free, were randomly divided into 8 groups (10 mice in each group) using random number table: reperfusion 2 h group, 6 h group, 12 h group, 24 h group, sham group, miR-330-3p agomir group (preoperative injection of agonist), miR-330-3p antagomir group (preoperative injection of inhibitor) and miRNA-NC group. Except for the sham group, the hepatic IRI model were established in mice. Polymerase chain reaction (PCR), Western blot and immunohistochemistry were used to detect the expression of miR-330-3p and phosphoglycerate mutase family member 5 (PGAM5), cleave caspase-1 and GSDMD. Luciferase reporter assay was performed to investigate whether miR-330-3p directly targets PGAM5. At the same time, AML12 cells were also treated with miR-330-3p mimics/inhibitor or PGAM5 siRNA, then the expression of PGAM5, NLRP3, cleave caspase-1 and GSDMD were detected by Western blot analysis.Results:Level of miR-330-3p gradually decreased after reperfusion, however, mRNA level of PGAM5 was increased thereafter ( P<0.05) as compared with the sham group. Serum level of AST and ALT were decreased in miR-330-3p agomir group while that of were increased in miR-330-3p antagomir group as a function of time following reperfusion, and the differences were statistically significant (all P<0.05). Cleave caspase-1 expression was decreased in miR-330-3p agomir group but was increased in miR-330-5p antagomir group ( P<0.05). Luciferase reporter assay was performed to determine PGAM5 was a target gene of miR-330-3p. SiRNA-mediated knockdown of PGAM5 decreased level of GAM5 (0.24±0.09), NLRP3(0.12±0.07), cleave caspase-1 (0.15±0.07) and GSDMD (1.08±0.08) as compared with the siRNA-NC group (1.17±0.14), (0.36±0.09), (0.68±0.09), (1.36±0.08), and the differences were statistically significant (all P<0.05). Conclusion:MiR-330-3p can alleviate hepatic IRI in mice, which may be related to inhibition of PGAM5-induced pyroptosis.

Objective To study the effect of subarachnoid hemorrhage (SAH) on voltagedependent potassium channels (Kv) currents of smooth muscle cells,which is hypothesized to be different between cerebral pial arteries and penetrating arteries.Methods Smooth muscle cells from cerebral pial arteries and penetrating arteries in rats were enzymatically isolated 72 h after SAH,and patch clamp was used to test the relative cell size,resting potential and Kv currents.Results Resting potential of either pial ((45.63 ±1.18) mV) or penetrating artery ((41.55-± 1.19) mV) was shifted positively by SAH,even more significantly in latter (F =8.24,P ＜ 0.05 ; F =9.36,P ＜ 0.01) ; Resting potential of pial artery of control ((38.76 ± 1.03) mV),penetrating artery of control ((38.53 ± 0.67) mV),pial artery of SAH ((36.87 ± 1.49) mV) and penetrating artery of SAH((37.89 ± 1.24) mV) were shifted positively to the same level by 1 μmol/L 4-aminopyridine (4AP; F =3.08,P ＞0.05).Maximum Current Density (Imax) of either pial ((20.82 ±0.59) pA/pF) or penetrating artery ((15.15 ±0.37) pA/pF) was compromised by SAH,also more significantly in latter (F =6.22,P ＜ 0.05) ; Imax of pial artery of control (9.15 ± 0.16),penetrating artery of control (9.04 ± 0.36),pial artery of SAH (8.77 ± 0.26) and penetrating artery of SAH (9.12 ± 0.17) were decreased to the same level by 1 μmol/L 4AP (F =2.96,P ＞ 0.05).Conclusions SAH probably shares the similar pathway with Kv blocker (4AP) in Kv currents inhibition.Further,SAH differently inhibits smooth muscle cells Kv currents and resting potential of cerebral pial arteries and penetrating arteries,which may be related with their different sensitivity towards cerebral vasospasm following SAH.

Objective To observe the specific changes of content of cyclic guanosine monophosphate (cGMP) around meridian skin and specific influences of electroacupunture Neiguan (PC6) on cGMP of rats with acute myocardial ischemia (AMI). Methods With 6 male Wistar rats as normal group, 18 rats were made AMI model and divided into model group, high-frequency group and low-frequency group (n=6). The high-frequency and low-frequency group were given electroacupuncture in bilateral PC6 with 100 and 2 Hz for 20 min respectively. The content of cGMP in skin around PC6, Ximen (PC4), Tianquan (PC2), Waiguan (TE5) and Zusanli (ST36) was detected by using enzyme-linked immunosorbent method after treatment. Results In model group, cGMP content increased significantly around PC6, PC2, TE5 and ST36 compared with normal group (P <0.05,P <0.001). After the treatment with high-frequency or low-frequency electroacupunture in PC6, cGMP content decreased significantly in skin around PC6, PC2, TE5 and ST36 compared with model group (P <0.05, P <0.001). Conclusion High-frequency electroacupunture or high-frequency electroacupunture used in PC6 can protect rats from the injury of AMI. The mechanism may be related to the decrease of cGMP content around the acupoint areas in heart-related meridian.

Objective To illustrate the composition ratio of ERβ isoforms in paired cancerous and adjacent normal tissues from breast cancer patients.Methods Eighty-seven pairs of cancerous and adjacent normal tissues were obtained from breast cancer patients.RT-qPCR was used to determine the relative mRNA expression levels of ERβ isoforms (ER[β1,ERβ2 and ERβ5),and the composition ratios of ERβ isoforms were analyzed.Results The expression levels of all tested ERβ isoforms (ERβ1,ERβ2 and ERβ5) in breast cancer tissues were much lower than those in adjacent normal breast tissues (P ＜ 0.01).Isoform ratio analysis showed that ERβ5 was the dominant isoform in both cancerous and adjacent normal tissues with a positive detection rate of 54.02 ％ and 75.84 ％,respectively.Meanwhile,ERβ1 had the lowest detection rate (9.74 ％ and 6.77 ％ in cancerous and adjacent normal tissues,respectively).The positive rates for both ERβ1 and ERβ2 were much lower in adjacent normal tissues than those in cancer tissues (Z =-2.24,P =0.025 and Z =-4.85,P ＜ 0.01,separately),while more cancerous tissues were ERβ5-positive in comparison to adjacent normal tissues (Z =-5.32,P ＜ 0.01).Conclusions The expression levels of all the ERβ isoforms are differentially down-regulated with significant alterations in their composition ratios during breast carcinogenesis.Further understanding on molecular mechanisms underlying the differential down-regulation of ER[β isoforms will shed new light on breast carcinogenesis.

10.3969/j.issn.1000-3606.2013.06.005

Mass spectrometry imaging (MSI), a label-free molecular imaging technique, has been applied widely in the spatial localization of small molecule metabolites, lipids, peptides, and proteins, with its unique advantage of high spatial resolving power compared to traditional liquid chromatography-mass spectrometry (LC-MS).With the nonstop advancement of its achievable sensitivity and spatial resolution, MSI technique has been providing novel perspectives into the preclinical studies of drugs, such as in vivo localization of drugs and their metabolites, visualization of drug metabolism, and drug delivery tracking.This review introduces the basics of MSI techniques, including basic principles, key features, technical advantages, and limitations, with particular highlight of the recent applications of MSI in drug efficacy and safety evaluation, drug distribution research, drug delivery research, and analysis of Chinese medicine from recent publications, aiming to promote the utilization and further expansion of MSI in the research and development of drugs.

Objective According investigate the expression of NLRP3 in liver tissues of mice with hepatic ischemia-reperfusion injury (HIRI),to determin the role of NLRP3 in the process of HIRI.Methods Established mice model of partial HIRI.Forty-two male C57BL/6 mice (aged 7 to 8 weeks,weight 20 to 25 g) were respectively divided into 7 groups:no-treatment control group,sham operation group,HIRI groups (2、6、12、24 h) and CY-09 group,6 mice in each group.The injury of the hepatic tissues in the 7 groups was analyzed based on detecting the levels of alanine transaminase (ALT),aspartate transaminase (AST),interleukin-1β (IL-1β),interleukin-18 (IL-18),tumor necrosis factor-α (TNF-α) by ELISA.HE and TUNEL staining were used to observe the pathological changes of liver tissues after HIRI.Western blotting assay were carried out to detect the expressions of NLRP3 and Caspase-1.Measurement data were expressed as mean ± standard deviation (Mean ± SD),and one-way variance analysis was used for comparison between groups.If the variance was not uniform,Dunnett C test was used.Results Serum ALT,AST,IL-1 β,IL-18 and TNF-α of mice detected in HIRI groups were higher than no-treatment control group and sham operation group at all endpoints (P ＜ 0.05).The relative expression of NLRP3 and Caspase-1 in the liver tissues of mice in the HIRI groups were significantly higher than that in the no-treatment control group and sham operation group.Serum ALT,AST,IL-1β,IL-18 and TNF-α of mice detected in CY-09 group were lower than HIRI groups at all endpoints (P ＜ 0.05).Less hepatocellular necrosis were exhibited in CY-09 group,comparing to HIRI groups.The hepatocyte apoptosis rate of mice in the CY-09 group was significantly lower than that in the 12 h HIRI group (P ＜ 0.05).The relative expression of NLRP3 in the liver tissues of mice in the CY-09 group was significantly lower than that in other groups.The relative expression of Caspase-1 in the liver tissues of mice in the CY-09 group was significantly lower than that in other groups except the no-treatment control group and sham operation group.Conclusions HIRI cause an increase in NLRP3 expression.The inhibition of NLRP3 can reduce HIRI.