Effect of miR-125b on T Cell Activation in Aplastic Anemia by Targetting B7-H4 / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of miR-125b on T cell activation in patients with aplastic anemia (AA) and its molecular mechanism.@*METHODS@#A total of 30 AA patients were enrolled in department of hematology, Binzhou Medical University Hospital from January 2018 to October 2021, as well as 15 healthy individuals as healthy control (HC) group. Peripheral blood mononuclear cells (PBMCs) were isolated, in which the levels of miR-125b and B7-H4 mRNA were detected by RT-qPCR. Immunomagnetic beads were used to separate naive T cells and non-naive T cells from AA patients and healthy people to detect the levels of miR-125b and B7-H4 mRNA. Lentivirus LV-NC inhibitor and LV-miR-125b inhibitor were transfected into cells, and T cell activation was detected by flow cytometry. The dual-luciferase reporter gene assay was used to detect the targetting relationship between miR-125b and B7-H4. RT-qPCR and Western blot were used to detect the levels of miR-125b, CD40L, ICOS, IL-10 mRNA and B7-H4 protein.@*RESULTS@#Compared with HC group, the expression of miR-125b was up-regulated but B7-H4 mRNA was down-regulated in PBMCs of AA patients (P <0.05), and the proportions of CD4+CD69+ T cells and CD8+CD69+ T cells in PBMCs of AA patients were higher (P <0.05). The expression of miR-125b was significantly up-regulated but B7-H4 mRNA was down-regulated in both naive T cells and non-naive T cells of AA patients (P <0.05), and non-naive T cells was more significant than naive T cells (P <0.05). Compared with NC inhibitor group, the expression of miR-125b was significantly decreased, the expression level of CD69 on CD4+ and CD8+ T cells in PBMCs was also significantly decreased, while the luciferase activity was significantly increased after co-transfection of miR-125b inhibitor and B7-H4-3'UTR-WT in the miR-125b inhibitor group (P <0.05). Compared with NC inhibitor group, the mRNA and protein levels of B7-H4 were significantly increased in the miR-125b inhibitor group (P <0.05). Compared with miR-125b inhibitor+shRNA group, the expression levels of CD69 on CD4+ and CD8+ T cells were significantly increased, and the levels of CD40L, ICOS and IL-10 mRNA were also significantly increased in the miR-125b inhibitor+sh-B7-H4 group (P <0.05).@*CONCLUSION@#MiR-125b may promote T cell activation by targetting B7-H4 in AA patients.

Analysis of surveillance results of Aedes albopictus in Hainan, 2017-2021 / 中国热带医学

@#Abstract: Objective　To understand the density changes and seasonal fluctuation of larvae and adults of Aedes albopictus in Hainan Province from 2017 to 2021, and to provide a scientific basis for the prevention and control of dengue fever and other vector-borne diseases in Hainan Province. Methods　The Breteau index method was used to monitor the larvae of Aedes mosquitoes, and the breeding sites of all small indoor and outdoor water containers and their larvae was checked and recorded. The adult mosquitoes were monitored by double lamination and light trap. Results　A total of 1 962 adult female Aedes albopictus mosquitoes were captured in Hainan Province from 2017 to 2021, accounting for 2.17% of the total captured mosquitoes. There were significant differences in mosquito species composition in different habitats in different years (χ2=312.258, P<0.01). In 2017, the density of mosquitoes in livestock sheds was the highest (9.71 per lamp·night), but the composition ratio of Aedes albopictus was the lowest (1.29%). In 2020, the mosquito density in hospitals was the lowest (2.22 mosquitoes per lamp·night), and in 2019, the proportion of Aedes albopictus in residential areas and hospitals was the highest (3.27% on average).The mosquito density in different habitats was the lowest in 2020. The proportion of Aedes albopictus in residential areas and livestock sheds showed an increasing trend from 2017 to 2021, while the proportion of other habitats showed significant fluctuations. The proportion of Aedes albopictus in hospitals showed an obvious decreasing trend in 2020. From 2017 to 2021, the average mosquito density was 5.97, 4.26, 3.82, 3.39 and 3.68 mosquitoes per lamp·night, respectively, showing a decreasing trend year by year except 2021. Except for 2019, the lure index also showed a decreasing trend, with an average value of 1.5/(hour·net), showing a decreasing trend year by year. From 2017 to 2021, the annual average density of Aedes albopictus in Hainan Province was 0.09 mosquitoes per lamp·night, showing obvious seasonal fluctuation. The Breteau index showed a slow decline from 2017 to 2021, and its annual peak was between June and August, which was 1-2 months later than the peak of adult mosquito density. Conclusions　The population density of Aedes albopictus in Hainan shows obvious seasonal characteristics. Mosquito control activities should be carried out according to the seasonal characteristics of mosquitoes to reduce the risk of dengue fever and other infectious diseases.

Effect of Nrf2/HO-1 signaling pathway in intestinal protection by Sishen Pills against ulcerative colitis in mice / 中国中药杂志

The present study aimed to explore the effect of nuclear factor erythroid 2 related factor 2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway in intestinal protection by Sishen Pills against ulcerative colitis(UC). After the UC model was induced by 3% dextran sodium sulfate(DSS), experimental animals were randomly divided into control group, model group, salazosulfapyridine(SASP) group, and low-and high-dose Sishen Pills groups. Drug intervention(ig) was performed for seven consecutive days during modeling. On the 7 th day, the mice were euthanized. The body weight and colon length were recorded, and the histopathological changes of the colon were observed by HE staining. Serum interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), total antioxidant capacity(T-AOC), malondialdehyde(MDA), and reactive oxygen species(ROS) were detected by ELISA. The protein and mRNA expression of Nrf2, HO-1, and NADPH quinine oxidoreductase-1(NQO-1) was determined by Western blot and reverse transcription-polymerase chain reaction(RT-PCR). Compared with the normal group, the model group exhibited reduced body weight, colon length, and T-AOC, increased IL-6, TNF-α, MDA, and ROS, and diminished protein and mRNA expression of Nrf2, HO-1, and NQO-1 in the colon tissues. Compared with the model group, the SASP group and high-dose Sishen Pills group showed elevated body weight, colon length, and T-AOC, lowered IL-6, TNF-α, MDA, and ROS levels, and increased protein and mRNA expression of Nrf2, HO-1, and NQO-1 in the colon tissues. As assessed by HE staining, Sishen Pills could improve the pathological changes of the colon. The findings suggested that Sishen Pills could protect the colon against UC induced by 3% DSS. The specific mechanism of action may be related to the anti-inflammatory and anti-oxidative stress effects by the activation of the Nrf2/HO-1 signaling pathway.

Property-efficacy Relationship of Scutellariae Radix and Atractylodis Rhizoma in Drying Dampness in Rats with Spleen-stomach Dampness-heat Syndrome / 中国实验方剂学杂志

Objective:To observe the difference in drying dampness between Scutellariae Radix and Atractylodis Rhizoma in model rats with spleen-stomach dampness-heat syndrome and clarify their property-efficacy relationship. Method:Sixty-four healthy male SD rats were randomized into the blank group， model group， high-， medium-， and low-dose Scutellariae Radix groups， as well as high-， medium-， and low-dose Atractylodis Rhizoma groups. The rats were exposed to high-fat and high-sugar diet and external dampness-heat environment for 20 days for inducing the spleen-stomach dampness-heat syndrome. The macroscopic manifestations of rats were observed and the morphological changes in stomach and colon were detected under a light microscope after hematoxylin-eosin （HE） staining， followed by the calculation of pathological scores. The serum tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-4 （IL-4）， and interferon-<italic>γ</italic> （IFN-<italic>γ</italic>） levels were determined by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA expression levels of aquaporin-4 （AQP4） in the gastric tissue were measured by Western blot and Real-time polymerase chain reaction （Real-time PCR）， respectively. Result:Rats in the model group presented with the manifestations of dampness-heat syndrome. The inflammatory reaction in stomach and colon was obvious， and the pathological score was significantly increased （<italic>P</italic><0.01）. The serum IFN-<italic>γ</italic>， IL-4， and TNF-<italic>α</italic> levels were elevated （<italic>P</italic><0.05）， and so were the AQP4 protein and mRNA expression levels in the gastric tissue except that there was no statistical difference. The clinical symptoms of rats in the medication groups were alleviated. Scutellariae Radix significantly relieved the gastric and colonic inflammation in model rats. Atractylodis Rhizoma inhibited the colonic inflammation in model rats to a certain extent， but it had no obvious effect on gastric inflammation. The pathological score of each Scutellariae Radix group was decreased. In terms of the pathological score of gastric tissue， only the high-dose Scutellariae Radix produced a significant difference （<italic>P</italic><0.01）， and the pathological scores of the three Atractylodis Rhizoma groups were not significantly different from that in the model group. As for the pathological score of colonic tissue， all the medication groups except for the low-dose Atractylodis Rhizoma group exhibited a significant difference in comparison with that of the model group （<italic>P</italic><0.01）. Scutellariae Radix and Atractylodis Rhizoma at each dose reduced not only the serum IFN-<italic>γ</italic>， IL-4， and TNF-<italic>α</italic> levels （<italic>P</italic><0.05， <italic>P</italic><0.01）， but also the AQP4 protein expression in gastric tissue of model rats （<italic>P</italic><0.01）. The AQP4 mRNA expression in the gastric tissue of model rats declined in the high- and low-dose Scutellariae Radix groups， while that in the medium-dose Scutellariae Radix group and each Atractylodis Rhizoma group rose without statistical difference. Conclusion:Scutellariae Radix and Atractylodis Rhizoma display a certain property-efficacy relationship in drying dampness of rats with spleen-stomach dampness-heat syndrome. Specifically， the efficacy of drying dampness is related to their cold/heat property， and the resulting outcome of bitter-cold Scutellariae Radix is better than that of bitter-warm Atractylodis Rhizoma.

Inhibitory Effect of Duanteng Yimu Decoction on Pannus Formation in Mice with Collagen-induced Arthritis Based on VEGF Pathway / 中国实验方剂学杂志

Objective:To investigate the mechanism of Duanteng Yimu decoction （DTYM） in the inhibition of pannus formation in collagen-induced arthritis （CIA） mice. Method:Twenty-four SPF-grade DBA/1 male mice were randomly divided into the following four groups： a blank group （NC group）， a model group （CIA group）， a methotrexate group （MTX group）， and a DTYM group， with six mice in each group. The mice， except for those in the NC group， were modeled. From the second immunization， the medium， MTX （1 mg·kg^-1）， and DTYM （15.4 g·kg^-1） were administered at an equal volume by gavage for 35 days. Mice were observed for general condition and the arthritis index. The knee and ankle joints were scanned by microcomputed tomography （micro CT）. Hematoxylin-eosin （HE） and safranin O/fast green staining were performed to observe pathological changes. Immunohistochemistry was performed to detect the expression of platelet/endothelial cell adhesion molecule-1 （CD31）， vascular endothelial growth factor-<italic>α</italic> （VEGF-<italic>α</italic>）， vascular endothelial growth factor receptor 2 （VEGFR2）， and phosphorylated（p）-VEGFR2. Result:Compared with the NC group， the CIA group showed red and swollen ankle joints， increased arthritis index scores （<italic>P</italic><0.05， <italic>P</italic><0.01）， manifest injury in the knee and ankle joints， reduced cartilage thickness， elevated Micro CT bone destruction scores of knee and ankle joints （<italic>P</italic><0.01）， and up-regulated absorbance values of synovial CD31， VEGF-<italic>α</italic>， VEGFR2， and p-VEGFR2 （<italic>P</italic><0.01）. Compared with the CIA group， the DTYM group showed relieved ankle joint redness and swelling， reduced arthritis index scores of mice three weeks after administration （<italic>P</italic><0.05， <italic>P</italic><0.01）， intact joint surfaces of the knee and ankle joints， thickened cartilage， declining Micro CT bone destruction scores in both the knee and ankle joints （<italic>P</italic><0.05， <italic>P</italic><0.01）， and lowered absorbance values of CD31， VEGF-<italic>α</italic>， VEGFR2， and p-VEGFR2 in the synovium （<italic>P</italic><0.01）. Conclusion:DTYM can inhibit the pannus formation in CIA mice presumedly by regulating the VEGF pathway.

Mechanism of Duanteng Yimu Decoction on Human Umbilical Vein Endothelial Cell Model / 中国实验方剂学杂志

Objective:To explore the effect of Duanteng Yimu decoction （DTYM） on the activation of the human umbilical vein endothelial cell （HUVEC） model and the effect on related activated proteins and vascular endothelial growth factor （VEGF） signaling pathway. Method:After DTYM （200， 400 g·mL^-1） treatment of HUVEC induced by VEGF and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， cell proliferation， migration， and tubulogenesis were detected by cell counting kit-8 （CCK-8） assay， 5-ethynyl-2'-deoxyuridine （EdU） assay， transwell migration assay， phalloidin staining， and matrix gel card method. The mRNA expression of adhesion factors， including E-selectin， intercellular adhesion molecule-1 （ICAM-1）， and vascular cell adhesion molecule-1 （VCAM-1） was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of von Willebrand factor （VWF）， platelet-endothelial cell adhesion molecule-31 （CD31）， angiogenic factor cysteine-rich-61 （CYR61）， angiopoietin-1 （ANG-1）， VEGF， and VEGF receptor-2 （VEGFR2） was detected by Western blot. Immunofluorescence was used to determine CD31 expression. Result:Compared with the normal group， the model group showed potentiated proliferation， migration， and tubulogenesis of HUVEC （<italic>P</italic><0.05， <italic>P</italic><0.01）， elevated mRNA expression of E-selectin， ICAM-1， and VCAM-1 （<italic>P</italic><0.01）， up-regulated protein expression of VWF， CD31， ANG-1， CYR61， VEGF-<italic>α</italic>， and phospho （p）-VEGFR2 （<italic>P</italic><0.05，<italic> P</italic><0.01）， and increased CD31 immunofluorescence intensity （<italic>P</italic><0.01）. Compared with the model group， the DTYM groups displayed blunted proliferation， migration， and tubulogenesis of HUVEC （<italic>P</italic><0.05，<italic> P</italic><0.01）， decreased mRNA expression of E-selectin， ICAM-1， and VCAM-1 （<italic>P</italic><0.05， <italic>P</italic><0.01）， down-regulated protein expression of VWF， CD31， ANG-1， CYR61， VEGF-<italic>α</italic>， and p-VEGFR2 （<italic>P</italic><0.05， <italic>P</italic><0.01）， and weakened CD31 immunofluorescence intensity （<italic>P</italic><0.01）. Conclusion:DTYM inhibits HUVEC proliferation， migration， adhesion， and tubulogenesis， which is associated with the regulation of CD31， VWF， CYR61， and ANG-1 expression in HUVEC and the VEGF signaling pathway.

Polymorphism of P66 in / 生物医学与环境科学（英文）

Objective@#To study the polymorphism in P66 and its human B-cell epitopes of @*Methods@#Polymerase chain reaction (PCR) and sequencing were used to obtain the P66 sequences of 59 Chinese @*Results@#Results showed that genetic and amino acid diversity presented in the 66 kD protein of all 59 Chinese strains, especially in @*Conclusion@#In P66 of 59 Chinese strains, polymorphisms were widely distributed. More importantly, the P66 amino acid sequences of

Bufalin-induced cardiotoxicity: new findings into mechanisms / 中国天然药物

Bufalin is one of the main pharmacological and toxicological components of Venenum Bufonis and many traditional Chinese medicine preparations. The cardiotoxicity clearly limits its application to patients living in countries. Hence, an investigation of its toxicological mechanism is helpful for new drug development and treatment of the related clinical adverse reactions. We investigate the cardiotoxicity of bufalin using human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) (0.003-0.1 μmol·L), human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) (0.03-0.3 μmol·L) and eight human cardiac ion channel currents (0.01-100 μmol·L) combined with an impedance-based bioanalytical and patch clamp method. Biphasic effect of bufalin on the contractility in hiPSC-CMs, which has been shown to strengthen myocardial contractility, accelerate conduction, and increase beating rate at the earlier stage of administration, whereas weakened myocardial contractility, abolished conduction, and ceased beating rate at the later stage of administration. Bufalin decreased the action potential duration (Action potential duration at 30%, 50% and 90% repolarization), cardiac action potential amplitude, and maximal depolarization rate and depolarized the resting membrane potential of hiPSC-CMs. Spontaneous beating rates of hiPSC-CMs were markedly increased at 0.03 μmol·L, while were weakened at 0.3 μmol·L after application. Bufalin blocks I in a concentration-dependent manner with half maximal inhibitory concentration of 74.5 μmol·L. Bufalin respectively increased the late sodium current and Na-Ca exchange current with a concentration for 50% of maximal effect of 2.48 and 66.06 μmol·L in hiPSC-CMs. Whereas, bufalin showed no significant effects on other cardiac ion channel currents. The enhancement of the late sodium current is one of the main mechanism for cardiotoxicity of bufalin.

Effect of Tongxie Yaofang with or Without Saposhnikoviae Radix on Water Metabolism and 5-HT Pathway in Irritable Bowel Syndrome Rats / 中国实验方剂学杂志

Objective:To investigate the effect of Saposhnikoviae Radix on Tongxie Yaofang in regulating water metabolism and 5-serotonin（5-HT） signal system in diarrhea type irritable bowel syndrome（IBS-D）rats. Method:The 40 IBS-D SD rats were randomly divided into model group, Tongxie Yaofang wituout Saposhnikoviae Radix group (26 g·kg-1), Tongxie Yaofang decoction group (30 g·kg-1), Tongxie Yaofang with double Saposhnikoviae Radix group (34 g·kg-1),another 10 normal SD rats were selected as the normal group.Except for normal group, the rats in other groups were separated from their mothers and induced by acetic acid to establish D-IBS model. These rats in the each group were administered with corresponding drugs for 14 days. The diarrhea indexs and the water contents of the feces were observed and calculated. The Na+-K+-ATPase activities in the intestinal mucosa were detected by micro method. The contents of 5-HT were detected by enzyme linked immunosorbent assay (ELISA). The activities of monoamine oxidase A (MAO-A) were detected by chemiluminescence method. The expressions of aquaporin 4 (AQP4) in colon were detected by immunohistochemistry. Protein expression levels of HT receptor 3 (5-HT3R), HT receptor 4 (5-HT4R) and serotonin transporter (SERT) in hypothalamus and colon were detected by Western blot. Result:Compared with normal group, the fecal water contents, contents of 5-HT in hypothalamus and colon, activities of MAO-A, expressions of 5-HT3R were significantly increased in model group (P<0.01), and the activities of Na+-K+-ATPase, expressions of AQP4, 5-HT4R and SERT were reduced significantly (P<0.01). Compared with model group, the diarrhea indexs and fecal water contents, contents of 5-HT in hypothalamus and colon, activities of MAO-A, expressions of 5-HT3R were significantly decreased in each experimental group, and the activities of Na+-K+-ATPase, expressions of AQP4,5-HT4R and SERT were significantly increased(P<0.05，P<0.01),however, there was no significant difference in the expression of 5-HT3R protein in Tongxie Yaofang wituout Saposhnikoviae Radix group.Compared with Tongxie Yaofang wituout Saposhnikoviae Radix group,the diarrhea index, fecal water content, 5-HT content, MAO-A enzyme activity, and protein expression of 5-HT3R were significantly decreased in Tongxie Yaofang group and Tongxie Yaofang with double Saposhnikoviae Radix group(P<0.05，P<0.01), and the activity of Na+-K+-ATPase and protein expression of SERT were significantly increased(P<0.05，P<0.01). Conclusion:Saposhnikoviae Radix can enhance the effects of Tongxie Yaofang on improving the water metabolism of rats with IBS-D and regulating the multi-target of 5-HT signaling system, further confirming the synergistic effect of Saposhnikoviae Radix.

Effect of Saposhnikoviae Radix on pharmacokinetics and tissue distributions of active components in Tongxie Yaofang in rats / 中国中药杂志

Tongxie Yaofang (TXYF) is a famous formula that has been used for treating gastrointestinal diseases in traditional Chinese medicine(TCM). Saposhnikoviae Radix is considered as a meridian guiding drug in TXYF and could enhance the effectiveness of prescription. However, the scientific evidence for this effect is still not clear. To reveal the interactions of Saposhnikoviae Radix with other herbs, we conducted this study on the pharmacokinetic profile and tissue distribution of active ingredients of TXYF in rats. The concentrations of four components in blood and tissues were determined by UPLC-MS/MS after oral administration with TXYF. The detection was carried out by electrospray ionization mass spectrometry in the multiple reaction monitoring (MRM) mode. The positive and negative ion switching technique was performed in the same analysis. The results revealed that Saposhnikoviae Radix could enhance Cmax, AUC0-t, and AUC0-∞ of paeoniflorin and hesperidin, and increase the distribution of atractylenolide-I, paeoniflorin and hesperidin in liver, spleen, brain and small intestine. Saposhnikoviae Radix increased the ratio of brain to blood concentrations of atractylenolide-I, paeoniflorin and hesperidin. Meanwhile, it reduced the ratio of lung to blood concentrations of atractylenolide-I and paeoniflorin. Saposhnikoviae Radix, and may enhance the effectiveness of prescriptions by promoting distribution of other herbs in brain.

Protective effect of dexmedetomidine against perioperative inflammation and on pulmonary function in patients undergoing radical resection of lung cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the protective effect of dexmedetomidine against perioperative inflammation and on pulmonary function in patients undergoing radical resection of lung cancer.</p><p><b>METHODS</b>From May, 2014 to May, 2016, 124 patients with lung cancer receiving radical surgeries were randomized into experimental group (n=62) and control group (n=62). The patients in the control group received a single anesthetic agent for anesthesia, and additional dexmedetomidine was given in the experimental group. The levels of serum interleukin-1β (IL-1β), IL-10, and tumor necrosis factor-alpha (TNF-α) were measured before the operation (T), at 30 min (T) and 60 min (T) during one lung ventilation (OLV) and at the end of operation (T). Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of malondialdehyde (MDA), myeloperoxidase (MPO) and xanthine oxidase (XOD), and the arterial oxygen partial pressure (PaO), oxygenation index (OI), airway plateau pressure (APP) and airway resistance (AR) were also recorded.</p><p><b>RESULTS</b>At the time points of Tand T, IL-1β, IL-10, MDA, MPO, TNF-α, and XOD levels were significantly increased in both of the groups, but the levels of IL-1, IL-10, TNF-α and MDA were significantly lower and MPO and XOD levels significantly higher in the experimental group than in the control group (P<0.05). In both groups, PaOand OI decreased and APP and AR increased significantly at Tand T, but APP and AR were significantly lower and PaOand OI significantly higher in the experimental group than in the control group (P<0.05).</p><p><b>CONCLUSION</b>Anesthesia with dexmedetomidine in lung cancer patients undergoing radical surgery can effectively reduce the inflammatory response of the lungs and protect the lung function of the patients.</p>

Effects of live combined bifidobacterium, lactobacillus and enterococcus powder on IgE and interleukin-17 levels in atopic children with bronchiolitis / 中华实用儿科临床杂志

Objective To observe the effects of live combined bifidobacterium,lactobacillus and enterococcus powder on immunoglobulin E (IgE) and interleukin-17 (IL-17) in atopic children with bronchiolitis.Methods Sixty cases of atopic children with bronchiolitis were randomly divided into the therapy group (30 cases) and the control group (30 cases).Twenty-five healthy children were enrolled as the healthy control group.Both the therapy group and the control group were given traditional therapy.The therapy group received live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months.The change of IgE and IL-17 levels were observed during the acutestage,remission stage and after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2months.Results (1) The levels of IgE and IL-17 of therapy group[(132.36 ±9.50) μg/L and (77.76 ±7.95)μg/L] during acute stage were markedly higher than those in the healthy control group [(52.80 ±4.92) μg/L and (46.92 ±4.79) μg/L] (all P ＜0.001).The levels of IgE and IL-17 of control group [(128.83 ± 8.06) μg/L and (76.61 ±6.18) μg/L] during remission stage were markedly higher than those in the healthy control group [(52.80 ±4.92) μg/L and (46.92 ± 4.79) μg/L] (all P ＜ 0.001).(2) The levels of IgE of therapy group (56.67 ± 9.20)μg/L after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months were markedly lower than those in the control group (70.50 ± 11.38) μg/L (P ＜ 0.001).The levels of IL-17 of therapy group [(49.63 ± 6.35) μg/L] at the time after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months were markedly lower than these in the control group (54.77 ± 6.33) μg/L (P =0.003).Conclusion Receiving live combined bifidobacterium,lactobacillus and enterococcus powder for two months can decrease the IgE and IL-17 levels in atopic children with bronchiolitis.

Evaluation of Six Recombinant Proteins for Serological Diagnosis of Lyme Borreliosis in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>In this study, we evaluated the diagnostic efficiency of six recombinant proteins for the serodiagnosis of Lyme borreliosis (LB) and screened out the appropriate antigens to support the production of a Chinese clinical ELISA (enzyme-linked immunosorbent assay) kit for LB.</p><p><b>METHODS</b>Six recombinant antigens, Fla B.g, OspC B.a, OspC B.g, P39 B.g, P83 B.g, and VlsE B.a, were used for ELISA to detect serum antibodies in LB, syphilis, and healthy controls. The ELISA results were used to generate receiver operating characteristic (ROC) curves, and the sensitivity and specificity of each protein was evaluated. All recombinant proteins were evaluated and screened by using logistic regression models.</p><p><b>RESULTS</b>Two IgG (VlsE and OspC B.g) and two IgM (OspC B.g and OspC B.a) antigens were left by the logistic regression model screened. VlsE had the highest specificity for syphilis samples in the IgG test (87.7%, P<0.05). OspC B.g had the highest diagnostic value in the IgM test (AUC=0.871). Interactive effects between OspC B.a and Fla B.g could reduce the specificity of the ELISA.</p><p><b>CONCLUSION</b>Three recombinant antigens, OspC B.g, OspC B.a, and VlsE B.a, were useful for ELISAs of LB. Additionally, the interaction between OspC B.a and Fla B.g should be examined in future research.</p>

Effect of Huanshuai Recipe Oral Liquid (characters: see text) on renal dysfunction progression in patients with atherosclerotic renal artery stenosis / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Huanshuai Recipe Oral Liquid ([characters: see text], HSR) on retarding the progression of renal dysfunction in patients with atherosclerotic renal artery stenosis (ARAS).</p><p><b>METHODS</b>A total of 52 ARAS patients with the Chinese medicine (CM) syndrome of qi deficiency and blood stasis, phlegm and dampness retention were recruited and randomly assigned into the treatment group (36 cases) and the control group (16 cases). Both groups received a basic treatment (high-quality low-protein diet, blood pressure control, lipid-lowering, correcting the acidosis, etc.). In addition, the treatment group received 20 mL HSR and the control group received placebo, 3 times a day for 6 months. Renal function (serum creatinine, blood urea nitrogen and uric acid) and blood lipids (cholesterol, triglycerides and low density lipoprotein) were examined monthly. The estimated glomerular filtration rate (eGFR) and CM syndrome score were compared between groups.</p><p><b>RESULTS</b>After treatment, compared with the control group, the serum creatinine level, uric acid level and CM syndrome score of the treatment group were significantly decreased (P<0.05 or P<0.01), and the eGFR in the treatment group were significantly increased (P<0.05).</p><p><b>CONCLUSION</b>HSR can effectively improve the renal function and clinical symptoms of ARAS patients.</p>

Combination of Loop-Mediated Isothermal Amplification Assay and Nested PCR for Detection of Borrelia burgdorferi sensu lato in Human Serum Samples / 生物医学与环境科学（英文）

A set of universal loop-mediated isothermal amplification (LAMP) primers targeting the fla gene was designed to detect Borrelia burgdorferi sensu lato (B. burgdorferi s.l.) in human samples. The sensitivity of LAMP was 20 copies/reaction, and the assay did not detect false positives among 11 other related bacteria. A positive LAMP result was obtained for 9 of the 24 confirmed cases and for 12 of 94 suspected cases. The positive rate of LAMP was the same as that of nested PCR. The LAMP is a useful diagnostic method that can be developed for rapid detection of B. burgdorferi s.l. in human sera. Combination of the LAMP and nested PCR was more sensitive for detecting B. burgdorferi s.l. in human serum samples.

Hypercoagulable state in patients with multiple myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to detect the plasma thrombomodulin (TM), D-dimer and fibrinogen in patients with multiple myeloma (MM) and to analyze their relationship with morbid state, and also to investigate the relationship of the expression of coagulation factor with the ratio of myeloma cells.</p><p><b>METHODS</b>ELISA was used to detect the TM level in 45 cases of MM at different stages. The plasma level of D-dimer and fibrinogen was detected by STA automatic coagulation analyser.</p><p><b>RESULTS</b>The level of plasma TM in newly diagnosed patients was higer than that in normal control group and in platform stage group (P < 0.01; P < 0.05). There were significant differences between relapsed or refractory group and normal control group or those reached platform stage group (P < 0.05). Meanwhile, the level of plasma TM in the group of thalidomide combined with chemotherapy was higer than that in newly diagnosed patients (P < 0.05). The level of plasma D-dimer and fibrinogen of MM patients was higher than that in normal controls (P < 0.01;P < 0.05). The expression of D-Dimer in relapsed or refractory group reached the maximum. Also, the level of plasma D-Dimer in group of thalidomide combined chemotherapy was higer than in newly diagnosed patients (P < 0.05). The expression of coagulation factor did not correlate with the ratio of myeloma cells.</p><p><b>CONCLUSIONS</b>Level of plasma TM, D-Dimer and fibrinogen of MM patients is higher than that in control group. The level of plasma TM and D-Dimer can be elevated when thalidomide used, which indirectly suggested the tendency for thrombosis in MM patients.</p>

Establishment of multiple locus variable-number tandem repeat analysis assay for genotyping of Borrelia burgdorferi sensu lato detected in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Human Lyme Borreliosis (LB), which is caused by Borrelia burgdorferi sensu lato (B. burgdorferi), has been identified as a major arthropod-borne infectious disease in China. We aimed to develop a multiple locus variable-number tandem repeat (VNTR) analysis (MLVA) assay for the genotyping of Borrelia burgdorferi strains detected in China.</p><p><b>METHODS</b>B. garinii PBi complete 904.246 kb chromosome and two plasmids (cp26 and lp54) were screened by using Tandem Repeats Finder program for getting potential VNTR loci, the potential VNTR loci were analyzed and identified with PCR and the VNTR loci data were analyzed and MLVA clustering tree were constrcted by using the categorical coefficient and the unweighted pair-group method with arithmetic means (UPGMA).</p><p><b>RESULTS</b>We identified 5 new VNTR loci through analyzing 47 potential VNTR loci. We used the MLVA protocol to analyse 101 B. burgdorferi strains detected in China and finally identified 51 unique genotypes in 4 major clusters including B. burgdorferi sensu stricto (B.b.s.s), B. garinii, B. afzelii, and B. valaisiana, consistent with the current MLSA phylogeny studies. The allele numbers of VNTR-1, VNTR-2, VNTR-3, VNTR-4, and VNTR-5 were 7, 3, 9, 7, and 6. The Hunter-Gaston index (HGI) of five VNTR loci were 0.79, 0.22, 0.77, 0.71, and 0.67, respectively. The combined HGI of five VNTR loci was 0.96. Clustering of the strains of Xinjiang, Inner Mongolia and Heilongjiang was confirmed, and this situation was consistent with the close geographical distribution of those provinces.</p><p><b>CONCLUSION</b>The MLVA protocol esytablished in this study is easy and can show strains' phylogenetic relationships to distinguish the strains of Borrelia species. It is useful for further phylogenetic and epidemiological analyses of Borrelia strains.</p>

Seroepidemiological investigation of lyme disease and human granulocytic anaplasmosis among people living in forest areas of eight provinces in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Lyme disease and Human granulocytic anaplasmosis are tick-borne diseases caused by Borrelia burgdorferi and Anaplasma phagocytophilum respectively. We have investigated infection and co-infection of the two diseases in the population of forest areas of eight provinces in China by measuring seroprevalence of antibodies against B. burgdorferi and A. phagocytophilum.</p><p><b>METHODS</b>Forest areas in 8 provinces were chosen for investigation using whole sampling and questionnaire survey methods. 3 669 serum samples from people in the forest areas were tested for the presence of antibodies by indirect immunofluorescent assay (IFA).</p><p><b>RESULTS</b>Seroprevalence against B. burgdorferi was 3% to 15% and against A. phagocytophilum was 2% to 18% in the study sites in the 8 provinces in China. We also found co-infection of B. burgdorferi and A. phagocytophilum in 7 of the 8 provinces (the exception being the Miyun area in Beijing). The seroprevalence for both B. burgdorferi and A. phagocytophilum was significantly higher among people exposed to ticks than among people who were not exposed to ticks.</p><p><b>CONCLUSION</b>We conclude that both pathogens are endemic in the forest areas in the eight provinces, but the prevalence of B. burgdorferi and A. phagocytophilum differs between the provinces.</p>

A study of the technique of western blot for diagnosis of lyme disease caused by Borrelia afzelii in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure.</p><p><b>METHODS</b>FP1, which is the representative strain of B. afzelii in China, was analyzed by SDS-PAGE, electro transfer and immunoblotting assays. The molecular weights of the protein bands of FP1 were analyzed by Gel-Pro analysis software. In a study using 451 serum samples (159 patients with Lyme disease and 292 controls), all observed bands were recorded. The accuracy of the WB as a diagnostic test was established by using the ROC curve and Youden index.</p><p><b>RESULTS</b>Criteria for a positive diagnosis of Lyme disease were established as at least one band of P83/100, P58, P39, OspB, OspA, P30, P28, OspC, P17, and P14 in the IgG test and at least one band of P83/100, P58, P39, OspA, P30, P28, OspC, P17, and P41 in the IgM test. For IgG criteria, the sensitivity, specificity and Youden index were 69.8%, 98.3%, and 0.681, respectively; for IgM criteria, the sensitivity, specificity and Youden index were 47%, 94.2%, and 0.412, respectively.</p><p><b>CONCLUSION</b>Establishment of WB criteria for B. afzelii is important in validating the diagnostic assays for Lyme disease in China.</p>

Optimization of pulse-field gel electrophoresis for Borrelia burgdorferi subtyping / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To optimize the performance of Pulsed-Field Gel Electrophoresis (PFGE) for the comparison of inter-laboratory results and information exchange of Borrelia burgdorferi subtyping.</p><p><b>METHODS</b>A panel of 34 strains of B. burgdorferi were used to optimize PFGE for subtyping. In order to optimize the electrophoretic parameters (EPs), all 34 strains of B. burgdorferi were analyzed using four EPs, yielding different Simpson diversity index (D) values and the epidemiological concordance was also evaluated.</p><p><b>RESULTS</b>The EP of a switch time of 1 s to 25 s for 13 h and 1 s to 10 s for 6 h produced the highest D value and was declared to be optimal for MluI and SmaI PFGE of B. burgdorferi. MluI and SmaI were selected as the first and second restriction enzymes for PFGE subtyping of B. burgdorferi according to discrimination and consistency with epidemiological data.</p><p><b>CONCLUSION</b>PFGE can be used as a valuable test for routine genospecies identification of B. burgdorferi.</p>