OBJECTIVETo study the effect of Buyang Huanwu decoction (BYHWD) inducing angiogenesis on the neuroblast migration from the subventricular zone and its mechanisms after focal cerebral ischemia.

METHODThe middle cerebral artery occlusion (MCAO) was performed to mice for 30 minutes to establish the model. The rats were divided into sham group, model group, BYHWD group and endostatin group. BYHWD (20 g x kg(-1), ig) and endostatin (10 μg, sc) were administered 24 h after ischemia once a day for consecutively 14 days. At 14 d after ischemia, the density of micro-vessel and the number of neuroblasts in the ischemia border zone were determined by immunofluorescence staining. The mRNA and protein expression of cell-derived factor-1 (SDF-1) and brain-derived neurotrophic (BDNF) were examined by real-time PCR and Western blot.

RESULTCompared with the model group, BYHWD significantly increased the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), and significantly increased the SDF-1 and BDNF mRNA and protein expression (P < 0.01). Compared with BYHWD group, endostatin significantly reduced the density of micro-vessel and the number of DCX positive cells in the ischemia border zone (P < 0.01), as well as the SDF-1, BDNF mRNA and protein expression (P < 0.01).

CONCLUSIONBYHWD could promote the neuroblast migration from the subventricular zone via inducing angiogenesis after cerebral ischemia, the mechanism may be correlated with up-regulating the expression of SDF-1 and BDNF.

Angiogenesis Inducing Agents ; pharmacology ; Animals ; Brain Ischemia ; pathology ; physiopathology ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Cell Movement ; drug effects ; Cerebral Ventricles ; pathology ; Chemokine CXCL12 ; analysis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Neurons ; drug effects ; physiology

OBJECTIVETo explore the effect of ligustrazine on the migration of bone marrow mesenchymal stem cells (BMSCs) and protein expressions of matrix metalloproteinase-2 and-9 (MMP-2 and MMP-9) in vitro.

METHODSBMSCs were in vitro isolated and cultured using whole bone marrow adherent method, and phenotypes [surface positive antigens (CD29 and CD90) and negative antigens (CD34 and CD45)] identified using flow cytometry. BMSCs were divided into the blank control group, 25, 50, 100 µmol/L ligustrazine group, and the GM6001 group (100 µmol/L ligustrazine +MMPs inhibitor GM6001 ). The migration of BMSCs was tested by Transwell chamber test and wound healing assay after treated with ligustrazine for 24 h. The protein expressions of MMP-2 and MMP-9 were detected by Western blot.

RESULTSThe third passage BMSCs grew well in uniform morphology. The expression rate of CD29, CD90, CD34, and CD45 was 96.9%, 97.3%, 0.2%, and 3.0%, respectively. Compared with the blank control group, the number of migrated cells and relative distance of cell invasion increased, and the protein expressions of MMP-2 and MMP-9 were elevated in each ligustrazine group (P < 0.05, P < 0.01). Compared with 100 µmol/L ligustrazine group, the number of migrated cells and relative distance of cell invasion decreased in 25 and 50 µmol/L ligustrazine groups and the GM6001 group (P < 0.01). Protein expression of MMP-2 decreased in 25 and 50 µmol/L ligustrazine groups (P < 0.01).

CONCLUSIONLigustrazine could promote the migration of BMSCs in vitro, and its mechanism might be related to up-regulating expression levels of MMP-2 and MMP-9 protein.

Cell Movement ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Pyrazines ; pharmacology ; Up-Regulation

The establishment of high specificity and sensitivity method of small molecule monoclonal antibody-based immunoassay has a great importance in the study of small molecule compounds in Chinese medicine, wherein synthesis of small molecule artificial antigen is a critical step in the preparation of small molecule antibodies. Oxidation method using sodium iodide was used to synthesize immunogenic antigen (FRn-BSA) and coating antigen (FRn-OVA) of forsythin. UV spectroscopy and thin layer chromatography showed that forsythin was successfully conjugated with BSA and OVA. After immuned FRn-BSA, the mice could specifically produce anti-forsythin antibodies with titer up to 1:8 000, and the linear range was from 1 mg x L(-1) to 100 mg x L(-1). In this paper, the artificial antigen of forsythin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.
Animals ; Antibodies ; immunology ; Antigens ; chemistry ; immunology ; Bridged Bicyclo Compounds, Heterocyclic ; chemistry ; immunology ; Drugs, Chinese Herbal ; chemistry ; Furans ; chemistry ; immunology ; Male ; Mice ; Mice, Inbred BALB C

Tetramethylpyrazine (TMP), an effective component of traditional Chinese medicine Chuanxiong, is commonly used to resolve embolism. Its possible therapeutic effect against atherosclerosis has received considerable attention recently. Angiotensin II (Ang II) is highly implicated in the proliferation of vascular smooth muscle cells (VSMCs), resulting in atherosclerosis. The mechanisms of TMP in the proliferation of VSMCs induced by Ang II remain to be defined. The present study was aimed to study the effect of TMP on Ang II-induced VSMC proliferation through detection of nuclear factor-kappaB (NF-kappaB) activity and bone morphogenetic protein-2 (BMP-2) expression. Primary cultured rat aortic smooth muscle cells were divided into the control group, Ang II group, Ang II + TMP group and TMP group. Cells in each group were harvested at different time points (15, 30 and 60 min for detection of NF-kappaB activity; 6, 12 and 24 h for measurement of BMP-2 expression). NF-kappaB activation was identified as nuclear staining by immunohistochemistry. BMP-2 expression was observed through Western blot, immunohistochemistry and in situ hybridization. The results showed that: (1) Ang II stimulated the activation of NF-kappaB. Translocation of NF-kappaB p65 subunit from cytoplasm to nucleus appeared as early as 15 min, peaked at 30 min (P<0.01) and declined after 1 h. (2) TMP inhibited Ang II-induced NF-kappaB activation (P<0.01). (3) Ang II increased BMP-2 expression at 6 h but declined it significantly at 12 and 24 h (P<0.01). (4) BMP-2 expression was also kept at high level at 6 h in Ang II + TMP group but maintained at the normal level at 12 and 24 h. (5) There was no significant difference in NF-kappaB activation and BMP-2 expression between the control group and TMP group. These results indicate that TMP inhibits Ang II-induced VSMC proliferation through repression of NF-kappaB activation and BMP-2 reduction, and BMP-2 expression is independent of the NF-kappaB pathway. In conclusion, TMP has therapeutic potential for the treatment of atherosclerosis.
Angiotensin II ; antagonists & inhibitors ; Animals ; Atherosclerosis ; drug therapy ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; analysis ; antagonists & inhibitors ; Immunohistochemistry ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; NF-kappa B ; analysis ; antagonists & inhibitors ; Pyrazines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; analysis ; antagonists & inhibitors

[Objective]To investigate the effect of KIF23 gene expression on the proliferation,migration and invasion of human hepatocellular carcinoma HepG2 cells in vitro,and to explore the possible mechanism.[Methods]The KIF23 siRNA was transfected into HepG2 cells by lipofectamine 3000.The expression of KIF23 mRNA and protein in HepG2 cells was de-tected by qRT-PCR and Western blot.The effect of silencing KIF23 on the proliferation of HepG2 cells was studied by CCK-8 assay and plate clone formation assay.The tumor cell abilities of migration and invasion after transfection were measured by scratch assay and Transwell assay.The expression of protein kinase B(PKB/Akt)and phosphorylated Akt(p-Akt)protein in HepG2 cells transfected with KIF23-siRNA2 was detected by Western blot.[Results]KIF23-siRNA could effectively si-lence the expression of KIF23 mRNA and protein in HepG2 cells(P<0.01).The results of CCK-8 assay,plate clone forma-tion assay,scratch assay and Transwell assay demonstrated that the cell proliferation,migration and invasion ability of the KIF23-siRNA2 interference group were significantly inhibited,compared to the negative control group and the blank control group(P<0.05).The expression level of total Akt protein in HepG2 cells was not changed,but the expression level of phos-phorylated Akt protein was down-regulated(P<0.05).[Conclusions]KIF23 may promote the proliferation,migration and in-vasion of human hepatocellular carcinoma cells by activating Akt signal transduction pathway.KIF23 is expected to be a new target for gene therapy of hepatocellular carcinoma.

Objective To investigate the expression of interleukin-3 receptor alpha(CD123)on bone marrow cells in acute myelocytic leukemia(AML)and its clinical significances.Methods By means of Fluorescence-activated cell sorer(FACS)and semi-quantity reverse transcripition polymerase chain reaction(RT-PCR),the expression of IL-3R?(CD123+)protein on CD34+CD38-cells and mRNA in BMMNCs of 62 AML patients of Tianjin Medical University General Hospital from March 2008 to January 2009 and 12 normal controls were detected respectively;Then the correlation between IL-3R? and the clinical stages of AML were analyzed.Results CD34+CD38-CD123+/CD34+CD38-and IL-3R? mRNA in BMMNC of 33 deno-vo or relapsed AML patients were higher than those of control group(P

OBJECTIVEThe novel influenza A (H1N1) virus firstly detected in April 2009 in Mexico rapidly spread to many countries including the United States and Canada where humans were infected with the H1N1 virus and deaths were reported. The pandemic virus strain had never been detected in specimen of human beings and swine. It was so highly contagious and widely spread that threatened life of humans globally. This study aimed to analyze clinical data of hospitalized children patients with 2009 novel H1N1 influenza A virus infection confirmed by etiologic tests, and compared with that of seasonal influenza A.

METHODClinical manifestations, laboratory and therapy data from the hospitalized children were collected by designed case report form and analyzed. All patients were enrolled from Capital Institute of Pediatrics from January 2003 to 2010. There were 152 cases in seasonal influenza A group, which was composed of 100 boys and 52 girls. Other 93 boys and 86 girls formed 2009 novel influenza A group.

RESULTInfluenza A was dominate from 2003 to 2008 and the peak season was December and January, while the peak hospitalized time of 2009 novel H1N1 influenza was from November 2009 to January 2010. The median age of seasonal influenza group was 35 months, which was lower than that of novel influenza group (Z = -6.702, P<0.01). Besides, 80.9% of the patients in seasonal influenza group were infants, while the novel influenza A group was mainly composed of infants and pre-school children (chi2 = 40.725, P<0.01). The cases of both groups had influenza-like symptoms at onset and the most common presentations were fever and cough. The duration of fever was much longer in 2009 novel influenza group (Z = -7.173, P<0.01). Patients in two groups nearly had the same symptoms except cough was more frequently presented by novel influenza A group cases (chi2 = 4.109, P<0.05). In laboratory examination, the novel influenza group had more cases with abnormality in blood platelet, CRP, ALT, and CK-MB than that of seasonal influenza group (chi2 = 7.562, 17.245, 4.398, 6.217, P<0.01). Patients in novel influenza A group had more changes in electrocardiogram (chi2 = 24.461, P<0.01). More patients had common underlying medical condition in novel influenza groups than those in seasonal influenza group (chi2 = 12.553, P<0.01). Furthermore, the groups had different age distribution in underlying medical diseases (chi2 = 7.231, P<0.05). Children with 2009 novel H1N1 virus infection tended to catch pneumonia (chi2 = 8.661, P<0.01) and became the severe cases (chi2 = 10.595, P<0.01). They had much higher ICU admission rate (chi2 = 12.873, P<0.01) and longer hospital stay (Z = -2.764, P<0.01).

CONCLUSIONAs a new variant of influenza virus A, 2009 novel H1N1 influenza A had stronger pathogenicity. Children with underlying medical conditions had the high risk to be infected and developed severe manifestations.

Adolescent ; Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H1N1 Subtype ; Influenza A virus ; Influenza, Human ; epidemiology ; virology ; Male

Objective:To explore the surgical method and therapeutic effect of repairing digital tip defect with free flap of proximal perforating branch of proper palmar digital artery.Methods:From March, 2009 to January, 2021, 15 patients with soft tissue defects at the tip of 16 digits were repaired with free perforator flap of proper palmar digital artery. The flap was obtained from the ulnar side of an index finger, on both sides of a middle finger and on the proximal side of the radial side of the ring finger. The size of flaps was 1.8 cm × 1.2 cm - 4.5 cm × 2.2 cm. The flap carried dorsal branch of proper palmar digital nerve and 0.5-4.5 cm of arteriae digitales palmares propriae. The donor digital artery was re-anastomosed in 3 cases 3 digits, transferred and anastomosed in 2 cases and un-anastomosed in 10 cases 11 digits. The dorsal branch of the proper palmar digital nerve in the flap was anastomosed with the proper palmar digital nerve of the finger stump at the recipient site to restore the sensation of flap, and the donor sites at the wrist transverse stripes or elbow transverse stripes were directly sutured. Regular follow-up via outpatient visit, telephone or WeChat interviews was conducted to observe the appearance, sensation and recovery of the flap and finger joint function.Results:After surgery, the flaps and donor site skin grafts of 15 cases with 16 digits were all survived, with first stage healing. A 4 months to 12 years follow-up showed that the flaps were in good texture and full shape with TPD at 7 - 11 mm. The joint function of digits was recovered well, and there was no complaint about uncomfortable donor site. According to the Michigan Hand Function Questionnaire, all 15 patients were satisfied with the overall appearance and function of the hands. According to TAM evaluation standard, all the digits of 15 patients were in excellent.Conclusion:Free flap of the proximal perforating branch of proper palmar digital artery is an ideal in the repair of digital tip soft tissue defect, as it has the advantages of an anatomical constant vessel, hidden donor site, less trauma caused, simple flap resection and good therapeutic effect.

Objective:To explore the tricuspid annular plane systolic excusion(TAPSE) in children with left-to-right shunt after congenital heart disease surgery and to understand the early systolic function of right heart in thesepatient.Methods:From June 2018 to December 2018, a prospective study was conducted in 20 infants after repair of left-to-right shunt congenital heart disease, including 10 males(50%) and 10 females(50%) , aged from 1 to 12 months, with a median of 4.5(2.0, 6.8) months, a body mass of 3.0-9.0 kg with median of 6.0(3.7, 7.7) kg.On the first postoperative day, blood was taken from central venous for N-terminal pro-B-type natriuretic peptide(NT pro-BNP) test, TAPSE and left ventricular ejective fraction(LVEF) was measured by echocardiography.The effects of aortic occlusion time, cardiopulmonary bypass time, preoperative pneumonia and preoperative heart failure on TAPSE were compared. The relationship between TAPSE and heart rate, systolic pressure, central venous pressure, vasoactive drug score, endotracheal intubation time, detention time in intensive care unit, NT pro-BNP and LVEF after operation was analysed.Results:The aortic cross-clamping time was 15-87 minutes, with median 31(28, 50) minutes. The cardiopulmonary bypass time was 35-117 minutes, with an average of(68±22)minutes. The time of tracheal intubation was 4-117 hours, with an average of(50±35) hours. The stay time in CICU was 1-14 days, with a median of 5(2, 7) days.The LVEF was 0.18-0.66, with median 0.53(0.42, 0.57). The TAPSE was 2.0-10.0 mm, with an average of(5.2±2.0) mm. On the first day after operation, NT pro-BNP was 1 548-35 000 pg/ml, with an average of(9 446±8 130) pg/ml.TAPSE was negatively correlated with postoperative intubation time( r=-0.576, P= 0.007) and detention time in ICU( r=-0.765, P=0.000), and positively correlated with postoperative LVEF( r=0.461, P=0.041)( P<0.05). TAPSE was negatively correlated with heart rate( r=-0.303, P=0.193), central venous pressure( r=-0.425, P=0.062), vasoactive drug score( r=-0.418, P=0.067) and NT Pro BNP( r=-0.348, P=0.132), and positively correlated with systolic pressure( r=0.146, P=0.54), but there was no statistical significance in each item.Compared with patients with TAPSE≥5mm, the detention time and tracheal intubation time were longer than those TAPSE<5 mm, the central venous pressure and NT-pro BNP was higher than those TAPSE<5 mm( P<0.05), the difference was statistically significant, other indicators had no significant difference. Conclusion:It is simple and feasible to measure TAPSE by echocardiography in children after operation with left-to-right shunt congenital heart disease.TAPSE decreased postoperatively suggested that the function of right ventricle decreased at the early stage after surgery, and with left ventricle systolic function decreased, which eventually led to the increase of NT pro-BNP, the need for higher doses of vasoactive drug support, longer tracheal intubation time and the stay time in CICU.Attention should be paid to the right heart function of children after congenital heart surgery.

Objective To assess the adherence,immunologic and survival responses in HIV-infected patients receiving free antiretroviral therapy (ART). Methods All adult HIV-infected patients in Wenxi county who started antiretroviral treatment (ART) between 01 July 2001 and 31 December 2006 and aged above 18 years were included in this study. Epidemiological survey and laboratory tests were performed before,0.5 months after, 1 months after, 2 months after and every 3 months after initiation of ART to recognize the adherence, efficacy (CD4+ T cell counts) and survival to the regimens. Results The median follow-up time period was 16.5 months (Interquartile: 15.5-20.8 months). At baseline, the median of CD4+ T cell counts were 154 cells/μl (Interquartile: 81-212 cells/μl). Treatment was effective in most of the patients, the CD4+ T cell count of patients increased after the initiation of ART. The maximum increase was recorded at month 3, from the median of 154 cells/μl to 220 cells/μl (P<0.001) ,and thereafter the count remained stable. When comparing with patients with baseline CD4+ T cell count≥100 cells/μl, those with baseline CD4+ T cell count < 100 cells/μl showed a higher mean increase in the first three months of treatment. The cumulative probability rates of remaining alive were 0.94,0.88 and 0.87 at 3,12,24 months, respectively. In multivariate Cox's proportional hazard models, after adjustment for the type ofinitial regimens (NVP vs. EFV/IDV), CD4+T cell count of less than 50 cells/μl (vs. 50 cells/μl or more) was strongly associated with death hazard ratio 0.21 (95% CI:0.06-0.68). Conclusion Our data showed that ART was effective for improving immunologic response of adult patients with HIV/AIDS. CD4+ T cell count at initiation was associated with survival time in patients starting ART,suggesting that monitoring of CD4+ T count should be strengthened to early initiate antiretroviral therapy for HIV-infected patients.