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<p><b>OBJECTIVE</b>To study the postnatal changes in lymphocyte subsets in early preterm infants and the effect of perinatal factors on lymphocyte subsets.</p><p><b>METHODS</b>A total of 61 early preterm infants were enrolled. Flow cytometry was used to measure the absolute counts of lymphocytes and lymphocyte subsets at 1, 7, 14, and 28 days after birth, as well as at 6 months after birth for 17 of these early preterm infants. The effects of perinatal factors, such as antepartum use of hormone, intrauterine infection, gestational age at birth, and Ureaplasma urealyticum (UU) colonization, on lymphocyte subsets were analyzed.</p><p><b>RESULTS</b>The absolute counts of lymphocyte subsets except natural killer (NK) cells were lowest at birth, increased rapidly at 1 week after birth, and reached the levels in healthy infants at 6 months; the count of NK cells remained at a low level and increased significantly at 6 months after birth. Compared with those with a gestational age of <28 weeks, the early preterm infants with a gestational age of ≥28 weeks had significantly higher absolute counts of T cells, T helper (Th) cells, and NK cells at 7 days after birth, a significantly higher absolute count of T cells at 14 days after birth, and significantly higher absolute counts of lymphocytes and Th cells at 28 days after birth (P<0.05). Compared with the group not using hormone, the group using hormone showed a significantly higher absolute count of T cells at 7 days after birth and significantly higher absolute counts of lymphocytes and all subsets at 14 days after birth (P<0.05). There was no significant difference in lymphocyte subsets at 1 day after birth between the intrauterine infection and non-infection groups (P>0.05); the intrauterine infection group had significantly higher absolute counts of B cells at 7 and 14 days after birth than the non-infection group. Compared those without UU colonization, the infants with UU colonization had significantly higher absolute counts of lymphocytes, T cells, Th cells, and Ts cells at 1 day after birth and a significantly higher absolute count of B cells at 14 days after birth.</p><p><b>CONCLUSIONS</b>Early preterm infants have deficiencies in innate immune cells at birth and normal levels at about 6 months after birth. Various perinatal factors including antepartum use of hormone, gestational age at birth, intrauterine infection, and UU colonization have long-term effects on lymphocyte subsets in early preterm infants.</p>
Subject(s)
Female , Humans , Infant, Newborn , Male , Infant, Premature , Allergy and Immunology , Lymphocyte Subsets , Microbiology , Physiology , Ureaplasma urealyticumABSTRACT
OBJECTIVE: To optimize the formulation of the doxorubicin magnetic thermosensitive liposomes (DOX-MTSLs) was optimized by orthogonal design and evaluated its property. METHODS: DOX-MTSLs were prepared by reverse evaporation combined with (NH4)2 SO4-gradient method. The entrapment efficiency was determined by mini-column centrifugation-RP-HPLC method. The transform temperature was determined by differential scanning caliorimetric (DSC). The particle size and Zeta electric potential were measured by transmission electron microscopy (TEM) and the laser scattering particle size distribution analyzer. The in vitro release behavior of DOX-MTSLs at different temperatures and different time points were studied by dialyzer in constant temperature water and simulated the release regression equation at(41 ±0.5)°C. The heating effect of DOX-MTSLs was determined in the high alternation magnetic field(AMF). RESULTS: The optimum recipe of DOX-MTSLs was founded as DOX/DPPC of 1:20(m/m), DPPC/Chol of 4:1 (mol/mol), the concentration of magnetic fluid 15 mg · mL-1, pH value of 7.4. The average entrapment efficiency of three batches of DOX-MTSLs(100416,100507,100517 self-preparation) were(82.77 ±0.88) %, (83.03 ±1.38)% and (80.68 ±0.42)% (n=3). The average particle size of doxorubicin magnetic thermosensitive liposomes were 177.1 nm, and the polydispersity index(PDI) was 0.700. When the temperature increased from (25 ± 0.5)°C to (37 ± 0.5)°C, the in vitro drug release was very slow and incomplete (15.45% and 19.54%) even up to 10 h. However, at the phase transition temperature(Tm) 41°C, the in vitro drug release significantly reached 90.99% within 2 h and continued to release. The DOX-MTSLs was showed to be temperature dependent and the in vitro release model was fitting the kinetic equation of Higuchi at(41±0.5)°C. The heating temperature and balanceable temperature of DOX-MTSLs(Fe3O2 3.19 mg · mL-1) were respectively raised to 39.9°C in 10 min and to 56.7°C in 40 min in high frequency induction heating equipment of oscillation frequency 80 kHz and current 12A. CONCLUSION: The optimized conditions can be obtained with high entrapment efficiency, good thermosensitivity and perfect magnetic susceptibility to reach the expectation.
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<p><b>OBJECTIVE</b>To evaluate the effect of FLAMIGEL (hydrogel dressing) on the repair of residual burn wound.</p><p><b>METHODS</b>Sixty burn patients with residual wounds hospitalized in 6 burn units from November 2011 to May 2012 were enrolled in the multi-center, randomized, and self-control clinical trial. Two residual wounds of each patient were divided into groups T (treated with FLAMIGEL) and C (treated with iodophor gauze) according to the random number table. On post treatment day (PTD) 7 and 14, wound healing rate was calculated, with the number of completely healed wound counted. The degree of pain patient felt during dressing change was evaluated using the visual analogue scale (VAS). The mean numbers of wounds with score equal to zero, more than zero and less than or equal to 3, more than 3 and less than or equal to 6, more than 6 and less than or equal to 10 were recorded respectively. Wound secretion or exudate samples were collected for bacterial culture, and the side effect was observed. Data were processed with repeated measure analysis of variance, t test, chi-square test, and nonparametric rank sum test.</p><p><b>RESULTS</b>Wound healing rate of groups T, C on PTD 7 was respectively (67 ± 24)%, (45 ± 25)%, and it was respectively (92 ± 16)%, (72 ± 23)% on PTD 14. There was statistically significant difference in wound healing rate on PTD 7, 14 between group T and group C (F = 32.388, P < 0.01). Ten wounds in group T and four wounds in group C were healed completely on PTD 7, with no significant difference between them (χ(2) = 0, P > 0.05). Forty-two wounds in group T and seven wounds in group C healed completely on PTD 14, with statistically significant difference between them (χ(2) = 42.254, P < 0.01). Patients in group T felt mild pain during dressing change for 37 wounds, with VAS score higher than zero and lower than or equal to 3. Evident pain was observed in patients of group C during dressing change for 43 wounds, and it scored higher than 3 and less than or equal to 6 by VAS evaluation. There was statistically significant difference in mean number of wounds with different grade of VAS score between group T and group C (Z = -4.638, P < 0.01). Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, E. coli, Baumanii, and Staphylococcus epidermidis were all detected in both groups, but there was no statistical difference between group T and group C (χ(2) = 0.051, P > 0.05). No side effect was observed in either of the two groups during the whole trial.</p><p><b>CONCLUSIONS</b>FLAMIGEL can accelerate the healing of residual burn wounds and obviously relieve painful sensation during dressing change.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Bandages , Burns , Therapeutics , HydrogelsABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of up- or down-regulation of haemoxygenase 1 (HO-1) gene expression on intestinal mucosa injury induced by intra-abdominal hypertension (IAH).</p><p><b>METHODS</b>(1) Reproduction of rat model of up- or down-regulation of HO-1 gene expression. Twenty-four healthy adult Wistar rats were divided into Co-PP (HO-1 specific revulsive) 2.5 mg, Co-PP 5.0 mg, Sn-PP (HO-1 specific inhibitor) 2.5 mg, and control groups according to the random number table, with six rats in each group. Rats in groups Co-PP 2.5 mg and Sn-PP 2.5 mg were respectively given Co-PP 2.5 mg/kg and Sn-PP 2.5 mg/kg by intraperitoneal injection, once every 12 hours for 3 days. The rats in group Co-PP 5.0 mg were intraperitoneally injected with Co-PP 5.0 mg/kg, once a day for 3 days. The rats in control group were treated with equal volume of normal saline by intraperitoneal injection. All rats were sacrificed on post injection day (PID) 4, and intestinal mucosa tissues were collected for determination of HO-1 mRNA expression. Optimal dose of Co-PP was chosen for the following experiment. (2) The influence of up- or down-regulation of HO-1 gene expression on intestinal mucosa injury under IAH condition. Another 24 healthy adult Wistar rats were divided into control, IAH, Co-PP+IAH, and Sn-PP+IAH groups according to the random number table, with six rats in each group. The rats in groups Co-PP+IAH and Sn-PP+IAH were intraperitoneally injected with 2.5 mg/kg Co-PP and 2.5 mg/kg Sn-PP, once every 12 hours for 3 days. Equal volume of normal saline was intraperitoneally injected into the rats in control group, once every 12 hours for 3 days. Then, nitrogen gas pneumoperitoneum was used to establish the model of IAH in rats of the latter three groups on PID 4, with IAP at 20 mm Hg (1 mm Hg = 0.133 kPa) , and it was maintained for 2 hours. Puncture and intubation were performed in rats of control group without inflating nitrogen gas. Jejunal segment in the length of 10-15 cm was harvested for collecting intestinal mucosa tissues to determine the HO-1 mRNA expression and diamine oxidase (DAO) content. Serum obtained from portal vein blood was collected to determine the D-lactate, TNF-α, and IL-6 contents. Another jejunal segment in the length of 1-2 cm was harvested for histopathological examination. Data were processed with one-way analysis of variance and t test.</p><p><b>RESULTS</b>(1) The HO-1 mRNA expression in group Co-PP 2.5 mg was significantly higher than that in control and Co-PP 5.0 mg groups (with t values respectively 4.756, 3.175, P < 0.05 or P < 0.01). The HO-1 mRNA expression in group Sn-PP 2.5 mg was significantly lower than that in control group (t = 4.880, P < 0.01). The optimal dose of Co-PP for the following experiment was 2.5 mg/kg. (2) HO-1 mRNA expression in group Co-PP+IAH was 60 ± 5, and it was obviously higher than that of group IAH (49 ± 5, t = 3.811, P < 0.01) and control group (39 ± 4, t = 8.034, P < .001) . HO-1 mRNA expression was higher in group IAH than in control group (t = 3.826, P < 0.01). HO-1 mRNA expression in group Sn-PP+IAH was 29 ± 4, which was obviously lower than that of control group (t = 4.330, P < 0.01). The contents of DAO and D-lactate in group Co-PP+IAH were (0.52 ± 0.05) U/mL and (1.9 ± 0.6) mg/L, which were significantly lower than those in group IAH [(0.88 ± 0.06) U/mL and (4.3 ± 0.7) mg/L, with t values respectively 11.291, 6.376, P values all below 0.01], but still higher than those in control group [(0.34 ± 0.04) U/mL, (1.2 ± 0.5) mg/L, with t values respectively 6.886, 2.295, P < 0.05 or P < 0.01]. The contents of TNF-α and IL-6 were much lower in group Co-PP+IAH than in group IAH, but still higher than in control group (with t values from 3.781 to 18.557, P values all below 0.01). The contents of DAO, D-lactate, TNF-α, and IL-6 in group Sn-PP+IAH were all higher than those in the other 3 groups (with t values from 4.181 to 32.938, P values all below 0.01). Structure of epithelial cells from intestinal mucosa was intact and regularly arranged in rats of control group. Intestinal mucosal tissue was edematous, and the top of villi was anabrotic and necrotic in rats of group IAH. Compared with that of group IAH, the degree of intestinal mucosa injury was alleviated in rats of group Co-PP+IAH, while the pathology was aggravated in rats of group Sn-PP+IAH.</p><p><b>CONCLUSIONS</b>Up-regulation of HO-1 gene expression can ameliorate intestinal mucosa injury caused by IAH, thus protecting intestinal mucosa tissues.</p>
Subject(s)
Animals , Rats , Disease Models, Animal , Gene Expression Regulation , Heme Oxygenase (Decyclizing) , Metabolism , Intestinal Mucosa , Pathology , Intra-Abdominal Hypertension , Pathology , Rats, Wistar , Up-RegulationABSTRACT
<p><b>OBJECTIVE</b>To investigate the matrix metalloproteinase-9 (MMP-9) expression in vascular endothelial cells stimulated by the serum obtained from children with Kawasaki disease (KD) during the acute phase in the absence and presence of MMP-9 small interfering RNA (siRNA).</p><p><b>METHODS</b>MMP-9 siRNA plasmids were constructed and transduced into vascular endothelial cells (ECV-304) by liposomal transfection. ECV-304 were cultured in 6 different conditional media: KD serum + siRNA negative control, normal serum, KD serum + MMP-9 siRNA1 (pSilencer3.1-MMP1), KD serum + MMP-9 siRNA2 (pSilencer3.1-MMP2), KD serum + gamma-globulin, and KD serum. RT-PCR and Western blot were used to detect MMP-9 expression at mRNA and protein levels in ECV-304.</p><p><b>RESULTS</b>The mRNA and protein expression of MMP-9 in ECV-304 cultured with 10% serum from KD patients (2.49 +/- 0.03, 1.20 +/- 0.04) and KD serum + siRNA negative control plasmid (2.45 +/- 0.03, 1.15 +/- 0.03) were significantly higher than those cultured with 10% serum from normal control children (1.21 +/- 0.03, 0.52 +/- 0.03, respectively; all P < 0.01) and the increased MMP-9 expression could be significantly inhibited by MMP-9 siRNA1, MMP-9 siRNA2 and gamma-globulin (100 mg/ml, all P < 0.01).</p><p><b>CONCLUSIONS</b>The increase of MMP-9 expression in vascular endothelial cells induced by the serum from KD patients might take part in the formation of coronary artery lesions. Two customized MMP-9 siRNA plasmids (pSilencer3.1-MMP1 and pSilencer3.1-MMP2) can significantly inhibit both MMP-9 mRNA and protein expression.</p>
Subject(s)
Child , Humans , Cells, Cultured , Endothelial Cells , Metabolism , Matrix Metalloproteinase 9 , Genetics , Metabolism , Mucocutaneous Lymph Node Syndrome , Genetics , Metabolism , Plasmids , RNA, Small Interfering , Genetics , TransfectionABSTRACT
Objective To investigate the clinical significance of matrix metalloproteinase-9(MMP-9)in the pathogenesis of Kawasaki disease(KD)and its complications of coronary arterial lesions(CAL).Methods Twenty-three inpatients with KD were admitted in hospital from Jan.2005 to Dec.2006,whose venous blood samples were obtained during the acute and convalescent stage respectively,and CAL was detected with 2-Dechocardiography.Twenty-seven venous blood samples(12 febrile children and 15 normal physical examination children)were collected as healthy controls.Serum levels of MMP-9 were measured by enzyme-linked immunoadsorbent assay(ELISA).According to the results of echocardiography,children were divided into 2 groups:with CAL group(n=13)and without CAL group(n=10).Results During the acute stage,serum MMP-9 levels were significantly higher in KD than those in controls(Pa
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Objective To investigate the risk factors on children with coronary artery lesions (CAL) secondary to Kawasaki di-sease(KD). Methods One hundred and forty - five patients with KD from January 1999 to December 2001 were collected. Among them, 93 cases without CAL and 52 cases with CAL. The test results, therapeutic methods and prognosis were analyzed in two groups. Results The duration of fever was longer and mean value of erythrocyte sedimentation rate (ESR) higher in group with CAL than those m the group without CAL(P0.05).The incidence of CAL was 18.8% (18/96) in patients of IVIG treatment within 10 days from onset, and 69.4% (34/49) in patients without IVIG treatment (P