Abstract: At present, nucleic acid detection technology based on the PCR principle is commonly used to detect malaria parasites, the existing Plasmodium detection methods mainly include microscopy, antigen immunoassay, and nucleic acid detection,but due to the long detection time, high personnel and equipment requirements, and other shortcomings, its popularization, and application at the grassroots level are limited. What challenges previous Plasmodium detection methods are the lack of experienced professionals and advanced equipment at the grassroots as well as the requirement of rapid detection of large samples under extreme conditions. The isothermal amplification technology developed in recent years has potential application prospects due to its simplicity, rapidity, high sensitivity, and high specificity. This article attempts to review the principles, characteristics, and prospects of various isothermal amplification technologies, and on this basis, focuses on the introduction of recombinase polymerase amplification (RPA) and recombinase⁃aided isothermal amplification (RAA) assay technologies and proposes the use of such recombinant enzyme amplification technologies to achieve rapid and accurate diagnosis of common Plasmodium species possibility and imagination.

Clinical study of modified Ganmai Dazao decoction in the treatment of yang deficiency climacteric depression and observe the effects of modified Ganmai Dazao decoction on neuroendocrine system in patients with yang deficiency climacteric depression. 86 cases were randomly divided into treatment group treated with modified Ganmai Dazao decoction and control group treated with Deanxit. The curative effect was evaluated with Hamilton's depressive scale (HAMD) and pittsburgh sleep quality scale (PSQI) before and at the end of the two and four weeks of the treatment, the serum levels of serotonin (5-HT), norepinephrine (NE), estradiol (E2), follicle stimulating hormone (FSH), luteotropic hormone (LH) were detected before and after the four weeks of the treatment The results showed that the total effective power of treatment group was 88.4% and the total effective power of control group was 81.4% after four weeks interference, with insignificant difference between the two groups. After two and four weeks of the treatment, the score of HAMD decreased remarkably in both groups (P < 0.01), with insignificant difference between the two groups in same phase. After two and four weeks of the treatment, the total score of PSQI decreased remarkably in both groups (P < 0.05), with significant difference between the two groups after four weeks (P < 0.01). After four weeks of treatment, the serum levels of 5-HT and NE increased (P < 0.01), with insignificant difference between the groups. After four weeks of treatment, the serum levels of E2 increased obviously (P < 0.05), the levels of FSH decreased obviously (P < 0.05), the levels of LH decreased insignificant, with insignificant difference between two groups. This study indicates that modified Ganmai Dazao decoction has obvious therapeutic effects in the treatment of climacteric depression, and showed equivalent efficacy with Deanxit, and modified Ganmai Dazao decoction has better effect on improving the sleep quality in patients than Deanxit, the effect of improved clinical symptoms may be through adjusted levels of 5-HT, NE, E2, FSH and LH of climacteric depression.
Adult ; Depression ; blood ; drug therapy ; etiology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Menopause ; drug effects ; psychology ; Middle Aged ; Neurosecretory Systems ; drug effects ; metabolism ; Norepinephrine ; blood ; Phytotherapy ; Serotonin ; blood

AIM:To investigate the effect of suppression of mel18 gene on the differentiation of human acute myeloid leukemia cell line HL 60 induced by cinnamaldehyde ( CA) .METHODS:HL60 cells were treated with low con-centration of CA or all-trans retinoic acid (ATRA).shRNAmel18 vector and shRNALuc control vector were employed to package lentiviruses which were then used to infect HL 60 cells.The virus-infected HL60 cells were treated with low con-centration of CA , and ATRA was used as a positive control of differentiation-inducing agent .The differentiation markers on the cell surface and cell cycle of virus-infected HL60 cells were analyzed by flow cytometry .Western blot was used to deter-mine the expression of MEL18, cyclin D1 and p27, as well as the phosphorylation level of Akt .RESULTS:Low concen-tration of CA and ATRA increased the expression of granulocytic differentiation marker CD 11b on the HL60 cells, with the decreased expression of MEL 18 in the HL60 cells.The expression of MEL18 decreased in shmel18 virus-infected HL60 cells (shmel18-HL60 cells), but did not change in shLuc-HL60 cells.The expression of CD11b on shmel18-HL60 cells increased with G 1-phase arrest , which went even higher after treatment with CA .The phosphorylation level of Akt and the expression of cyclin D1 decreased in shmel18-HL60 cells with the increase in the expression of p27.CONCLUSION:In-hibition of mel18 gene leads HL60 cell granulocytic differentiation .mel18 gene may affect the differentiation of HL 60 cells by regulating cyclin D1 and p27 via PI3K/Akt signaling pathway.PI3K/Akt signaling pathway is also involved in CA-in-duced differentiation of HL60 cells by suppressing mel18 gene expression.

OBJECTIVETo develop a RP-HPLC method to determine plumbagin in Plumbago zeylanica, and to investigate contents of plumbagin in different parts of. P. zeylanica.

METHODThe analysis was carried out at 30 degrees C on a Kromasil C18, column eluted with a mobile phase consisting of a mixture of methanol-water (65: 35). The flow rate was 1 mL x min(-1), the detector wavelength was 213 nm.

RESULTThe calibration curve was linear within the concentration ranges of 0.020 8-0. 104 microg (r = 0. 9999). The average recovery was 98.7%. The contents in the root, stem and leaf were 0.394 5%, 0.050 8%, 0.031 4% respectively.

CONCLUSIONThis method is simple, accurate, replicate and suitable for the determination of plumbagin in P. zeylanica.

Chromatography, High Pressure Liquid ; methods ; Naphthoquinones ; analysis ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Plumbaginaceae ; chemistry ; Quality Control ; Reproducibility of Results

OBJECTIVETo establish the digitized visible human with Jiuwei (CV 15) involved.

METHODSWith the virtual Chinese human (VCH) datasets and three-dimensional modeling software adopted, the knowledge on acupuncture and moxibustion as well as acupoint anatomy combined and the computer image processing software applied, the visualized browser software of Jiuwei (CV 15) was established.

RESULTSBy establishing the interactive three-dimensional visualization browser software of acupoints, the location of Jiuwei (CV 15) was enabled to be expressed directly from all the levels, as well as the main adjacent tissue structure. It was suggested that Jiuwei (CV 15) should be punctured obliquely downward to avoid injuring the vital organs such as the heart and liver, and the safe depth of insertion should be 1.0-1.5 cm.

CONCLUSIONThe technology of digital visible human enables the three-dimensional expression of acupoint, which can be the platform of the digital teaching pattern. The research on the angle and depth of needling insertion at acupoint can be conducted in combination with teaching materials.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Human Body ; Humans ; Imaging, Three-Dimensional ; Software ; Therapy, Computer-Assisted ; instrumentation ; User-Computer Interface

OBJECTIVETo evaluate the neuroprotective effect of tetramethylpyrazine (TMP) against focal cerebral ischemic injury in rats with diffusion-weighted magnetic resonance imaging (DWMRI).

METHODSRat models of focal cerebral ischemic injury were established in 16 male SD rats. They were randomly divided into the TMP group and the control group, eight in each group, and pretreated with TMP and normal saline respectively before modeling. Change of infarcted cerebral focus was observed with DWMRI at 1, 2, 6, 12 and 24 hrs after infarction, and the infarction volume (IV) at 24 hrs after modeling was estimated by triphenyltetrazolium chloride (TTC) stain.

RESULTSThe IV in all time points observed in the TMP group with DWMRI was significantly smaller than that in the control group (P<0.01). Compared with that at 1 hr after infarction, in the control group at 2, 6, 12 and 24 hrs after modeling, the IV enlarged by 13.3%, 29.7%, 50.3% and 57.3% respectively, while that in the TMP group 9.9%, 21.3%, 37.1% and 40.5% respectively. The cerebral IV estimated by TTC stain 24 hrs after modeling was larger than that estimated by DWMRI.

CONCLUSIONTMP pretreatment before modeling was effective in protecting brain against cerebral ischemic damage in rats. DWMRI dynamic scanning observation has important significance in observing the cerebral ischemic developing process and evaluating the effectiveness of brain protective measures.

Animals ; Diffusion Magnetic Resonance Imaging ; Infarction, Middle Cerebral Artery ; drug therapy ; pathology ; Male ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Phytotherapy ; Pyrazines ; pharmacology ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley

The pharmacokinetics of 6beta-naltrexol (6beta-NOL) following single intramuscular administration and multiple intramuscular injection once per day for seven days was studied in 4 Beagle dogs. Plasma concentration of 6beta-NOL in dogs was analyzed by a combination of high performance liquid chromatography (HPLC) and electrochemical detection with naloxone (NLX) as internal standard. After single intramuscular injection of 0.2 mg x kg(-1) 6beta-NOL, the plasma concentration-time curve of the drug was found to fit to a two compartment model with first-order absorption. The main parameters of single dosing were as follows: t1/2alpha was (0.26 +/- 0.23) h, t1/2beta was (4.77 +/- 1.65) h, C(max) was (81.65 +/- 5.61) ng x mL(-1), t(peak) was (0.27 +/- 0.07) h, CL(s) was (1.20 +/- 0.06) L x kg(-1) x h(-1), V/F(c) was (1.94 +/- 0.15) L x kg(-1), and AUC(0-t) was (166.82 +/- 7.68) ng x h x mL(-1), separately. After multiple intramuscular injection of 0.2 mg x kg(-1) 6beta-NOL once per day for seven days, the plasma concentration-time curve of the drug fitted to a two compartment model with first-order absorption too. The main parameters of the last dosing were as follows: t1/2alpha was (0.19 +/- 0.18) h, t1/2beta was (5.79 +/- 1.50) h, C(max) was (79.82 +/- 10.5) ng x mL(-1), t(peak) was (0.18 +/- 0.08) h, CL(s) was (1.12 +/- 0.07) L x kg(-1) x h(-1), V/F(c) was (2.10 +/- 0.27) L x kg(-1), and AUC(0-t) was (173.23 +/- 9.49) ng x h x mL(-1), separately. The difference of the parameters between the first and the last dosing was not significant, showing that the plasma kinetics of 6beta-naltrexol was not changed after multiple administrations. In the course of multiple administration, the peak and valley concentration of plasma 6beta-naltrexol were (79.03 +/- 10.3) and (1.50 +/- 0.93) ng x mL(-1), respectively. No clear adverse events were noted during this study. These results showed that plasma 6beta-naltrexol fits to a two compartment model with first-order absorption in dog after intramuscular administration and their pharmacokinetic parameters were reported. There was no remarkable change on plasma pharmacokinetics of 6beta-naltrexol after multiple intramuscular administrations.
Animals ; Chromatography, High Pressure Liquid ; Dogs ; Half-Life ; Injections, Intramuscular ; Male ; Naltrexone ; administration & dosage ; analogs & derivatives ; pharmacokinetics

Objective To develop a cellular shear stress loading system with an adjustable stress mode and relevant parameters, and subsequently verify the effectiveness and feasibility of this system. Methods The hardware of the system was developed by using a peristaltic pump and self-designed multi-channel flow chamber, and the mode control program of shear stress based on LabVIEW was designed to control the device via RS485 interfacing and Modbus protocol. Additionally, the relationship between the shear stress and system parameters was calibrated, and finite element analysis was also conducted. Finally, the feasibility of the system was confirmed via the in vitro cell experiment. Results The mode and magnitude of shear stress of the system could be controlled via either the peristaltic pump or computer, and the cellular long-term effect was also able to be detected. The calibration results of the system indicated that the level of shear stress exhibited significantly linear positive correlation with the revolution of the peristaltic pump (P<0.001). Finite element analysis demonstrated that the level of shear stress on the slide was uniformly distributed and the result of simulation was accordant with calibration. Cytoskeleton staining suggested that cellular morphology of MLO-Y4 cells was changed, and microfilament increased and arrayed along fluid flow direction. Conclusion The system is stable and reliable enough to provide different loading modes and magnitude of cellular shear stress to offer a convictive platform of the research for different cellular stress signal transduction mecha-nisms.

The aim of this study was to investigate the apoptosis-inducing effect of cinnamic aldehyde (CA) on chronic myeloid leukemic (CML) cells and its mechanism. K562 cells and primary bone marrow mononuclear cells (MNC) from patients with CML were treated by various concentrations of CA. Flow cytometry was employed to measure the apoptosis of K562 cells and primary CML bone marrow MNC. Western blot was used to determine the expression of C-MYC and the phosphorylation of CrkL in K562 cells, and real-time polymerase chain reaction (real-time PCR) was used to quantify the expression of BCR-ABL mRNA in K562 cells. The results indicated that CA induced the apoptosis of K562 cells in a time- and dose-dependent manner. CA induced apoptosis of CML MNC dose-dependently. CA inhibited the expression of BCR-ABL mRNA and C-MYC, reduced CrkL phosphorylation levels in K562 cells. It is concluded that CA induces apoptosis of CML cells in vitro. Down-regulation of the expression and function of BCR-ABL may be one of its most important anti-leukemia mechanisms.
Acrolein ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Fusion Proteins, bcr-abl ; metabolism ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology

OBJECTIVETo explore the immune tolerance induction (ITI) in a severe hemophilia A patient with inhibitor, and to improve the therapeutic efficacy for patient.

METHODSThe FVIII:C was assayed by one-stage method and FVIII antibody by Bethesda method. Mutation screening of FVIII gene intron 22 inversion was performed using LD-PCR.

RESULTSFVIII gene intron 22 inversion was detected in this patient. Clinical tolerance to FVIII was successfully induced after administration of the ITI regimen combined with immunosuppression. A fall of inhibitor titer from 8 BU to 0 BU after treatment for 3 months, and in vivo FVIII recovery (> 66%) was normalized. The patient had no bleeding episode in the following 6 months.

CONCLUSIONThis is the first case report on successful immune tolerance induction therapy in Chinese hemophilia A patient. ITI is the most effective therapy for hemophilia A with inhibitor.

Autoantibodies ; immunology ; Factor VIII ; genetics ; Hemophilia A ; genetics ; Humans ; Immune Tolerance ; drug effects ; Immunosuppression