OBJECTIVE To develop new 2?5 type of piezoelectric immunoglobulin microarray immunosensor for determination of immunoglobulin.The energy converters are 10MHz AT-cut quartz crystals with gold-coated(electrodes).The monoclonal antibodies of immunoglobulin are immobilized onto the surfaces of crystals gold(electrodes) by staphylococcal protein A(SPA).METHODS The standard substance and serum were detected to find the detection time,temperature and specificity of the immunosensor.RESULTS The piezoelectric quartz(crystal) immunoglobulin microarray immunosensor could complete the detection in 15 min without nonspecific(response).Under the optimized conditions,the experimental results showed that the piezoelectric immunosensor had good(response) to immunoglobulin whose frequency shifts were linearly dependent on immunoglobulin (concentration) in different range.The piezoelectric microarray immunosensor had been used to detect(immunoglobulin) in serum,the analytical results given by this method were in satisfactory agreement with those given by traditional (immunoassay),with correlation coefficient of 0.94,0.94,0.94,0.92 and 0.91.(CONCLUSIONS) Piezoelectric microarray immunosensor for the determination of immunoglobulin is of high(sensitivity),high specificity,high analysis speed,unnecessary labelling,simple operation,real-time detection,(repeated) use,etc.It is suitable for (detecting) of immunoglobulin and should be used for clinical detection.

OBJECTIVE To have a research on the detection of Mycobacterium tuberculosis by fluorescence chip of molecular beacon probe.METHODS Fluorescence chip for the detection of M.tuberculosis was designed,based on the better bonding force and specificity of molecular beacon and target molecule.Hybridization efficiency was optimized,which included the concentration and purity of molecular beacon probe,hybridization temperature,hybridization time,the prescription of hybridization solution and the comparing of molecular beacon fluorescence signals of 4 molecules.RESULTS Molecular beacon fluorescence signal approached the highest when the final concentration of Mg was 5 mmol/L and the amplification efficiency of PCR approached the highest.Half extended molecular beacon probe could bind nice with chip when it was middle labeled with quench group.The results of 3 hybridization solutions were as follows: 0.15% SDS better than 0.2% SDS,10?SSC better than 5?SSC.Strong signal of hybridization reaction could be detected when the concentration of Cy3-BDH molecular beacon probe was 15 ?mol/L,the concentration of FAM-DABCLYE probe was 9 ?mol/L and the reaction time was 7 h.The efficiency of fluorescence quenching was higher when the molecule was consisted of Cy3-BDH,other than Cy3-DABCLYE.CONCLUSIONS The design of half extended molecular beacon probe tactfully makes use of the advantages of the high bonding force and specificity of molecular beacon probe on the chip technology.Fluorescence chip of molecular beacon for M.tuberculosis has satisfactory detecting rate,specificity and sensibility.

OBJECTIVE To develop a new type of piezoelectric immunoglobulin microarray immunosensor for(determination) of immunoglobulin.METHODS The energy converters were 10MHz AT-cut quartz crystals with gold-coated electrodes.The monoclonal antibodies of immunoglobulin were immobilized onto the surfaces of(crystals) gold(electrodes) by SPA to study the best immobilization yield of the antibodies at different condition.Gold-(protein) A complex was highly stable and was believed to be Van der Waals in nature.We did experiments to find the best temperature-time condition about the binding of SPA and the gold surface and to explore the influence(about) the immobilization of antibody by the concentration and the pH of the antibody solution.RESULTS At the same antibody concentration, it had the highest immobilization yield of SPA at a low-temperature-long-time(combination).The most immobilization yield was obtained at the 5mg/ml concentration of antibody solution.The optimum pH for antibody immobilization was 7.2.CONCLUSIONS SPA for the immobilization of antibody is of(simple) operation,and time-saving.It is a good way to immobilize antibody on the gold electric surface of the quartz(crystal).

OBJECTIVE To design molecular beacon probe of 43 Codon of high mutation site in rpsL of STR resistant MTB and its amplification system, meanwhile, to find the way of detecting the fluorescent light and making a qualitative judgment by use of fluorescence microscope and image analysis software. METHODS The software, Beacon designer, was used to design the 43 Codon molecular beacon probe and the amplification system, and the fluorescence microscope and image analysis software were used to detect the fluorescent light and make a qualitative judgment. RESULTS The strap of amplification product was shown clearly. The difference between PCR products from standard strain and STR resistant one was obvious in detecting the fluorescent light by use of fluorescence microscope. The rate of resistant STR detected was about 80% (P

OBJECTIVE: To investigate the changes of GATA-1 protein expression during erythroid differentiation of K562 cells under hypoxia and how GATA-1 can regulate erythroid differentiation by up-regulating the expression of miR-451a and inhibiting the expression of 14-3-3ζ. METHODS: K562 cells were divided into 2 groups: the normoxia group and the hypoxia group, after the induction of hemin for 96 h, the positive cells rate of the benzidine staining, the mRNA expression of γ-globin and the expression of CD235a were detected, and the success of the model was verified. The changes of GATA-1 and miR-451a expression in the above-mentioned 2 groups, the changes of miR-451a expression after over-expressed GATA-1 were detected by Western blot and qRT-PCR. The cells in normoxic group and hypoxia group were divided into negative control group (NC group) and miR-451a over-expression group respectively, and the degree of erythroid differentiation in the four groups was judged according to the corresponding erythroid differentiation indexes, and the expression of 14-3-3ζ was detected by Western blot after over-expressed miR-451a. RESULTS: The positive cell rate of benzidine staining, mRNA expression of γ-globin and the expression of CD235a after 96 h induction by K562 cells under hypoxia were significantly higher than 0 h, suggesting that the erythroid differentiation model of K562 cells under hypoxia was replicated successfully. The expression levels of GATA-1 protein and miR-451a in the hypoxic group were significantly higher than that in the normoxic group (P<0.05). The expression level of miR-451a in hypoxia group was significantly higher than that in NC group after overexpressed GATA-1 (P<0.05). After over-expressed of miR-451a under hypoxia, the positive cell rate of benzidine staining, the mRNA expression level of γ-globin and the expression of CD235a were significantly higher than those in NC group (P<0.05). The expression level of 14-3-3ζ protein in miR-451a over-expressed group was lower than that in NC group under hypoxia (P<0.05). CONCLUSION Hypoxia can significantly increase the expression of GATA-1 protein, and the increase of GATA-1 expression can up-regulate the expression of miR-451a, thereby inhibiting the expression of 14-3-3ζ protein, which hinders the cell proliferation in erythroid differentiation model of K562 cells and plays an important role in promoting erythroid differentiation.
14-3-3 Proteins ; Cell Differentiation ; Erythroid Cells/metabolism* ; GATA1 Transcription Factor/metabolism* ; Humans ; Hypoxia ; K562 Cells ; MicroRNAs/genetics*

Chronic hypoxic lung diseases are major causes of disability and mortality worldwide, which are typically aggravated by hypoxic pulmonary hypertension.The pathogenesis of hypoxic pulmonary hypertension is complex, and its mechanism has not been fully elucidated.The previous studies have shown abnormally elevated levels of free Ca + in the cytoplasm of pulmonary artery smooth muscle cells to be the predominant drivers of pulmonary hypertension , causing continuous contraction and remodeling of the pulmonary vessels.This article briefly summarizes the mechanism of hypoxia-induced imbalance in calcium homeostasis in pulmonary artery smooth muscle cells, together with its related drug research, based on the existing literature.Hypoxia induces an imbalance in calcium homeostasis in pulmonary artery smooth muscle cells by regulating hypoxia-inducible factor-1, K+ , store-operated calcium channel, receptor-operated calcium channel, the Ca +-sensing myosin contractile mechanism by binding to calmodulin, leading to pulmonary vasoconstriction.Ca + can also activate PKC/ MAPKs and PI3K/Akt/mTOR pathways, leading to pulmonary vascular remodeling.

OBJECTIVE: To investigate the clinical significance of bone marrow microvessel density(MVD) and angiogenesis related factors in multipic myeloma(MM). METHODS: Twenty cases of MM and 20 cases of simple fracture were selected and enrolled in MM group and control group respectively. The clinical data and results of laboratorial tests were collected; the bone marrow MVD of patients was detected by using the modified plastic-embedded pathologic sections of bone marrow tissue and histochemistry staining, the expression levels of amgiogenesis-related factors including VEGF, TNF-α, HGF, TGF-α, TGF-β1, bFGF, Ang-Ⅰ, Ang-Ⅱ in bone marrow supernatant were detected by ELISA; the mRNA expression levels of above-mentioned cytokines in bone marrow mononuclear cells were detected by real time-PCR; the pearson correlation analysis was used to analyze the correlation of MVD with VEGF, HGF and bFGF levels. RESULTS: The MVD in MM group was significantly higher than that in control group (P＜0.001); the mRNA expression of VEGF, TGF-α, TGF-β1 and HGF in bone marrow mononuclear cells of MM group was higher than that of control group(P＜0.001); the levels of VEGF, HGF, bFGF and THF-α in bone marrow supernatant of MM group were higher than those in control group(P＜0.05), moreover, the MVD positively correlated with levels of VEGF, HGF and bFGF in bone marrow(r=0.488, 0.472 and 0.457). CONCLUSION The MVD and levels vessel-related factors in bone marrow supernatant of MM patients increase, among which the levels of VEGF and HGF in bone marrow supernatant are consistant with those mRNA expression level in bone marrow mononuclear cells, moreover, the MVD possitively cerrelates with levels of VEGF, HGF and bFGF in bone marrow supernatant, suggesting that the changes of bone marrow microenvironment vassel-related factors play an important role in angiogenesis and pathogenesis of multiple myeloma.
Bone Marrow ; Bone Marrow Cells ; Humans ; Microvessels ; Multiple Myeloma ; Neovascularization, Pathologic ; Tumor Microenvironment

OBJECTIVE: To explore the pathogenesis of erythrocytosis by detecting the key enzymes of glucose metabolism and glucose transporter in bone marrow erythrocytes of chronic mountain sickness (CMS), and analyzing its correlation with hemoglobin. METHODS: Twenty CMS patients hospitalized in Qinghai Provincial People's Hospital from January 2019 to December 2020 were selected as CMS group. Twenty males with leukocyte count > 3.5×10⁹/L who had accepted bone marrow aspiration and had normal result were taken as control group. The mRNA and protein expression of key enzymes and glucose transporter in glucose metabolism in bone marrow CD71⁺ erythrocytes were detected by real time qPCR and Western blot, respectively. Glucose, lactic acid and 2,3-diphosphoglycerate in the bone marrow supernatant and serum were tested by ELISA. The mRNA and protein expression of key enzymes and glucose transporter, glucose, lactic acid and 2,3-diphosphoglycerate of the two groups were compared. Pearson correlation was used to analyze the correlation between key enzymes, glucose transporter in glucose metabolism in bone marrow CD71⁺ erythrocytes and hemoglobin. RESULTS: The expression of HK2, GLUT1 and GLUT2 mRNA in the CMS group were higher than those in the control group (P<0.001), while the expression of HK1, OGDH and COX5B mRNA were not different. The expression of HK2, GLUT1 and GLUT2 protein in the CMS group were higher than those in the control group (P<0.05). The levels of glucose and lactic acid in the bone marrow supernatant and serum in the CMS group were not different from those in the control group, while the level of 2,3-diphosphoglycerate was higher (P<0.001). Both HK2 and GLUT2 proteins were positively correlated with hemoglobin (r=0.511, 0.717). CONCLUSION CMS patients may increase glycolysis by increasing the expression of HK2, and promote the utilization of glucose through high expression of GLUT1 and GLUT2 to meet the need of energy supply.
Male ; Humans ; Altitude Sickness/metabolism* ; Glucose Transporter Type 1 ; 2,3-Diphosphoglycerate ; Hemoglobins ; Chronic Disease ; RNA, Messenger ; Phenotype ; Glucose

A total of 72 patients with first-episode mild-and-moderate depression took Qishen Fukang Capsules.The change of event-related potential P3 was determined before and after treatment for 6,12 and 24 weeks.And another 70 healthy controls received the same tests simultaneously.Compared with before treatment,HAMD scores after treatment were lower in the patient group [(29.1 ± 5.1) score vs.(10.4 ±4.1) score].P3 latencies were shorter and amplitudes became higher after treatment (P ＜0.01).And the difference of before and after treatment for 6 weeks was larger than the difference of 12 weeks and after treatment for 6 weeks (P ＜ 0.01).The curative effect is definite.And the clinical symptoms and brain evoked potential index have improved in patients with first-episode mild-and-moderate depression after medication.

Objective To investigate the method and effectiveness of the digital artery series and parallel of island flap for repairing degloved injuries of the fingertip of thumb or adjacent finger Methods Between September 2008 and July 2012,the finger artery series retrograde island flaps tiled in the repair of finger degloving injury in 13 cases,repair of adjacent finger tip degloving injury in 11 cases; 8 cases of degloved injuries of the fingertip were tiled with the digital artery parallel island flap from ulnaris middle finger and radialis ring finger of arteria digitalis communis pedicled which were from the same palm side.The size of skin and soft tissue defect ranged from 2.0 cm × 1.8 cm-7.2 cm ×5.5 cm,gutted flap minimum ranged from 1.1 cm × 1.0cm-1.5 cm × 1.3cm,and the maximum ranged from 3.0 cm × 2.2 cm-5.5 cm × 4.5 cm.The donor sites were repaired with the intermediate split thickness free skin grafts and performed with pressure dressing.Results The group of 32 cases,in 1 case the flap vasospasm occurred in operation of free process,the symptoms disappeared after local application of papaverine and hot compress ; 1 case of flap occurred disturbance of blood circulation after operation because of tight suturing for pedicle,the symptoms relieved after removing the stitches at intervals; 2 cases blisters appeared after operation,disappeared after a week.All skin flaps were survived,incision and skin graft donor sites healed by first intention.Twenty patients were followed-up from 6 to 12 months after operation.All flaps presented the satisfactory appearance and texture,recovered protective feeling.At last follow-up,the two-point discrimination was 7 to 10mm,and the flexion and extension function of wounded fingers recovered to normal.According to the functional assessment criteria of upper limb formulated by the Hand Surgery Branch of Chinese Medical Association,there were excellent in 19 cases,good in 10 cases,and moderate in 3 cases.No significant loss to the donor shape and function.Conclusion The surgery by adopted the digital artery series or parallel of island flap for repairing degloved injuries of the fingertip of thumb or adjacent finger,not only has the advantages of simpleness,safety and reliability,but also can satisfy the patients who aren't willing to accept or because of physical reasons can't accept the treatment of abdominal skin tube or nail flap from hallux toe,which performs in both high-end and low level hospitals,and deserves of general application.