Adenovirus-mediated Expression of B-domain-Deleted Human Coagulation Factor Ⅷ in ADSC of SD Rats / 中国实验血液学杂志

OBJECTIVE@#To transinfect SD adipose tissue-derived stem cell (ADSC) in vitro with a recombinant adenoviral vector containing human B-domain-deleted FVIII (BDDhFⅧ), so as to lay the foundation for the treatment of hemophilia A by using ADSC combined with BDDhFⅧ gene.@*METHODS@#ADSCs were isolated from the inguinal adipose tissue of SD rats and passed to third passage for identification. Third passage ADSCs were transfected in vitro with recombinant adenovirus vector Ad-BDDhFⅧ-GFP. The experiments were divided into Ad-BDDhFⅧ-GFP-transfected ADSCs group (A), Ad-GFP-transfected ADSC group (B), and untransfected ADSC group (C). CCK-8 method was used to detect the proliferation of transfected cells in three groups, and the expression level of hFⅧ antigen in cell supernatant was detected by ELISA. RT-PCR and Western blot respectively were used to detect the mRNA and protein expression of BDDhFⅧ in the three groups after transfection.@*RESULTS@#The growth curve of third passage cells isolated and cultured showed an inverted "S" shap; the flow cytometry detection showed the positive expression of CD29, CD90, CD44, and the negative expression of CD45 in third passage cells. After the adipogenic and osteogenic induction, the cells could transformed to adipogenic and osteogenic directions. CCK-8 detection showed that the proliferation of cells in 3 groups not was influenced. ELISA showed that the expression of hFⅧAg in group A was significantly higher than that in group B and C (P＜0.05). RT-PCR showed that compared with group A, there was no target band in B and C groups, and BDDhFⅧ gene was not expressed. The results in group A were consistent with the length of amplified fragments, and BDDhFⅧ target gene was expressed. Western blot analysis showed that the expression of hFⅧ protein in group A was significantly higher than that in group B and C. (P＜0.05).@*CONCLUSION@#Recombinant adenovirus Ad-BDDhFⅧ-GFP can effectively transfect rat ADSC in vitro, which lays an experimental foundation for gene therapy of hemophilia A.

A Preliminary Exploration on the Pathogenesis of Osteopenia in Patients with Hemophilia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the influencing factors and pathogenesis of osteopenia in the patients with hemophilia.</p><p><b>METHODS</b>Twenty-three patients with hemophilia were admitted in the hospital affiliated to North China University of Science and technology from March to August 2015, including 13 severe cases, 10 mild and moderate cases. All the patients accepted the detection of serum I collagen cross-linking N terminal peptide (NTX I), osteoprotegerin (OPG), bone alkaline phosphatase (BALP), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF) and transforming growth factor-β1 (TGF-β1), the score scale of activity ability was recorded according to the criteria published by the U.S. Center for disease prevention and control in 2002, and 21 patients received the measurement of bone mineral density. According to the World Health Organization (WHO) definition, the clinical significance of bone mineral density (BMD) was assessed by measuring the Z level.</p><p><b>RESULTS</b>Z level>-2 was recorded in 10 cases, Z≤-2 was recorded in 11 cases; the levels of body mass index (BMI) and human bone alkaline phosphatase (BALP) reflecting bone formation in 11 cases (Z≤-2) were lower than there in 10 cases (Z>-2) (P<0.05); the levels of BALP (r=0.489, P<0.05), IGF (r=0.538, P<0.05) and BMI (r=0.572, P<0.01) positively correlated significantly with BMD (P<0.05); the levels of bFGF (r=0.570, P<0.01) and OPG (r=0.505, P<0.05) positively correlated with NTX I, indicating bone destruction (P<0.05); the score of activity ability of severe patients was significantly lower than that of mild and moderate cases (P<0.05), BMD levels of these 2 groups were not statistically different (P>0.05).</p><p><b>CONCLUSION</b>The BMD level does not correlate with the clinial grouping of hemophilia, the low body mass index may be a risk factor for bone lose; the mechanism of hemophilia patient's bone lose may be related with the decrease of osteogenic activity, the IGF can prevent bone lose in hemophilia, the bFGF and OPG can promote bone metabolism of the patients with hemophilia.</p>

Correlation study on effects of electroacupuncture on the muscle condition and MNCV in rabbits with lumbar nerve root compression / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture on the muscle condition and electrophysiology of the muscle in rabbits with lumbar nerve root compression and to explore potential mechanisms.</p><p><b>METHODS</b>Twenty-four New Zealand white rabbits were randomly divided into a normal group, a model group, a medication group and an electroacupuncture group. The rabbit model with lumbar nerve root compression was established in the model group, the medication group and the electroacupuncture group. The model and the treatment were not produced in the normal group. The medication group was treated with oral administration of Loxoprofen tablets at a dose of 30 mg each day for 14 days. The electroacupuncture group was treated with electroacupuncture at "Jiaji" (EX-B 2) of L5 and L6 for 14 days. The hibateral muscle condition before and after making model and after treatment was detected and the electromyogram and the nerve conduction velocity (NCV) were observed.</p><p><b>RESULTS</b>The spontaneous potential and the insertion potential were increased in the model group, the medication group and the electroacupuncture group after making model. The value of the activation and the relaxed condition of the strength-creep (ARCSC) in the right muscle was significantly decreased. The motor nerve conduction velocity (MNCV) and the wave amplitude of evoked potential (WAEP) were significantly decreased in the model group, the medication group and the electroacupuncture group after treatment, but the value of MNCV and WAEP in the electroacupuncture group, and ARCSC in the right muscle in the electroacupuncture group and the medicine group were closed to those level in the normal group, among them, the relaxed condition in the electroacupuncture group was more close to the level in the normal group.</p><p><b>CONCLUSION</b>Electroacupuncture can improve the strength-creep condition of the muscle that damage nerve place controls and the electrophysiology of the muscle can recover the MNCV and the wave amplitude of evoked potential in the compression nerve root.</p>

Observation on therapeutic effect of simple obesity treated with acupuncture, auricular point sticking and TDP / 中国针灸

<p><b>OBJECTIVE</b>To probe into the mechanism and the best program of acupuncture for slimming.</p><p><b>METHODS</b>The patients were randomly divided into a control group (n = 60) treated with diet control and exercise, and a treatment group treated with acupuncture, auricular point sticking, TDP radiation. Changes of body weight, cholesterol (TC), triglyceride (TG) and blood sugar (BS) before and after treatment were investigated.</p><p><b>RESULTS</b>The effective rate of the treatment group (91.6%) was better than that of the control group (28.3%).</p><p><b>CONCLUSION</b>Acupuncture, auricular point sticking and TDP radiation combined with diet control and exercise are a better program for slimming, and decreasing appetite, promoting lipid decomposition metabolism are the mechanism of acupuncture in slimming.</p>