Protective effects of Salvia miltiorrhiza and Carthamus tinctorius active ingredients in different compatibility on cerebral ischemia/reperfusion injury in rats / 中草药

Objective To explore the protective effects of Salvia miltiorrhiza and Carthamus tinctorius active ingredients in different compatibility on cerebral ischemia-reperfusion injury in rats. Methods Adult male Sprague-Dawley (SD) rats were selected to establish a cerebral ischemia-reperfusion (I/R) injury model by middle cerebral artery occlusion (MCAO) method. The rats were randomly divided into sham operation group, model group, and positive control group (Danhong Injection 2 mL/kg); And orthogonal test method L9(34) was adopted to compose nine compatibility groups from main active ingredients of S. miltiorrhiza and C. tinctorius (trashinol, salvianolic acid A, salvianolic acid B, and HYSA). Rats were tail iv administrated once daily for continuous 3 d. Score neurological deficit was evaluated after 3 d; qRT-PCR was used to detect mRNA expression of glucose regulated protein 78 (GRP-78), nuclear factor-κB (NF-κB), C/EBP homologous protein (CHOP), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6); HE staining was used to detect the pathological changes of cerebral cortex, and the expression of NF-κB p65 protein in cerebral cortex was detected by immunohistochemistry. Results Compared to the model group, Danhong Injection group and orthogonal compatibility nine groups significantly reduced the neurological deficit scores of rats with cerebral ischemia-reperfusion; increased the mRNA expression of GRP-78 in the cerebral cortex; decrease the mRNA expression of NF-κB, CHOP, TNF-α, and IL-6, and decrease the protein expression of NF-κB p65 in the cerebral cortex. The results also showed that the protective effects of the 4th group (danshensu 30 mg/kg, salvianolic acid A 2.5 mg/kg, salvianolic acid B 16 mg/kg, and HYSA 8 mg/kg), 6th group (trashinol 30 mg/kg, salvianolic acid A 10 mg/kg, salvianolic acid B 8 mg/kg, and HYSA 4 mg/kg) in the 9th group were more significant for cerebral ischemia reperfusion injury in rats. Conclusion The active components of S. miltiorrhiza and C. tinctorius can play a good protective role in endoplasmic reticulum stress and inflammation in rats with cerebral ischemia reperfusion injury.

Preparation and evaluation of intra-articular injectable sinomenine hydrochloride-loaded in situ liquid crystals / 药学学报

Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64 : 16 : 20, w/w/w) loaded with 6 mg x g(-1) of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8 : 3.2 : 16 : 20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC(0-∞) of ISH2 group were increased significantly compared with that of SMH solution group and the AUC(0-∞) of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.

Preparation and evaluation of intra-articular injectable sinomenine hydrochloride-loaded in situ liquid crystals / 药学学报

Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64:16:20, w/w/w) loaded with 6 mg·g-1 of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8:3.2:16:20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC0-∞ of ISH2 group were increased significantly compared with that of SMH solution group and the AUC0-∞ of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.

Molecular diagnosis of OTC gene mutation in a Chinese family with ornithine transcarbamylase deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutations of OTC gene in a male infant affected with ornithine transcarbamylase deficiency.</p><p><b>METHODS</b>Genomic DNA were isolated from peripheral blood samples of family members and 100 healthy individuals. Potential mutations of the 10 exons of OTC gene were screened with PCR and Sanger sequencing.</p><p><b>RESULTS</b>A homozygous missense mutation c.917G>C in exon 9, which results in p.R306T, was identified in the infant. Sequencing of the mother and two female members of the family indicated a heterozygous status for the same mutation. The same mutation was not found in other members of the family and 100 healthy controls.</p><p><b>CONCLUSION</b>A missense mutation c.917G>C in the OTC gene is responsible for the pathogenesis of the disease. Identification of the mutation can facilitate prenatal diagnosis and genetic counseling for the family.</p>

Role of the Th17/Treg functional imbalance on the development of atherosclerosis in apo E knockout mice / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the role of the helper T cells (Th) 17/Treg cell imbalance on the development of atherogenesis in apo E knockout mice.</p><p><b>METHODS</b>Apo E(-/-) mice were examined at age of 6, 12, 24 and 48 weeks (n = 10 each). Age matched C57/B6 mice served as controls. The number of Th17, Treg and dendritic cell (DC) was detected by flow cytometry. The levels of interleukin(IL)-6, IL-17A and transforming growth factor(TGF)-β1 were detected by ELISA. The suppression ability of Treg was evaluated by mixed lymphocyte reaction.</p><p><b>RESULTS</b>With increasing ages, the frequencies of Th17 and Treg in CD4(+) T cells were increased (Th17 ratio from 1.00% to 3.14%; Treg ratio from 8.08% to 27.80%) and the level of IL-17A was up-regulated [from (87 ± 15) pg/ml to (191 ± 26) pg/ml], but the rate of Th17/Treg cell and the level of TGF-β1 remained stable during atherogenesis in apo E knockout mice. Furthermore, the phenotype of splenic DC was matured and the blood level of IL-6 was up-regulated [from (43 ± 5) pg/ml to (104 ± 11) pg/ml] with aging in apo E(-/-) mice. Addition of IL-6 to T cells reversed the ability of Treg to suppress the proliferation of effective T cells.</p><p><b>CONCLUSION</b>DC overactivation, subsequent increased secretion of IL-6, inhibition of Treg cell function and the Th17/Treg cell imbalance play key roles on the atherogenesis in apo E(-/-) mice.</p>

Association between AA-NAT gene polymorphism and reproductive performance in sheep

Arylalkylamine N-acetyltransferase (AA-NAT) is critical enzyme in Melatonin (MLT) biosynthesis for MLT regulating the animal seasonal breeding. In this study, DNA sequencing methods were applied to detect the polymorphisms of the AA-NAT gene in 179 Chinese sheep belonging to two non-seasonal reproduction breeds and two seasonal reproduction breeds. One mutation at exon 3 (NM_001009461:c.486A > G) was firstly described at the sheep AA-NAT locus. Hence, we described the SmaI PCR-RFLP method for detecting EX3 486A > G mutation, frequencies of the AA-NAT-G allele varied from 0.871 to 0.908 in two non-seasonal reproduction breeds and 0.517 to 0.578 in two seasonal reproduction breeds. The associations of SmaI polymorphism with estrus traits was analyzed in non-seasonal reproduction breeds sheep and seasonal reproduction breeds sheep, the significant statistical results were found between them, the GG genotype frequencies was higher in non-seasonal reproduction breeds (p < 0.001), while, the GA genotype frequencies was higher in seasonal reproduction breeds (p < 0.05). Hence, the EX3 486A > G mutation could facilitate association analysis and serve as a genetic marker for Chinese sheep breeding and genetics.

ASS1 mutation leading to citrullinemia I in a Chinese Han family / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate potential mutation of the ASS1 gene in a male infant with acute citrullinemia type I.</p><p><b>METHODS</b>Genomic DNA was prepared from peripheral blood samples of the family members. Mutation analysis of the 14 ASS1 exons was carried out by PCR and direct DNA sequencing.</p><p><b>RESULTS</b>A homozygous missense mutation of c.970G>A located in exon 13, which results in p.G324S, was identified in the child. Sequencing of the parents showed a heterozygous status for the same mutation.</p><p><b>CONCLUSION</b>A missense mutation of c.970G>A in the ASS1 gene is responsible for the pathogenesis of the disease in the infant.</p>

Effects of maternal deficiency of folic acid during pregnancy on pulmonary development and SP-A expression in newborn rats / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study examined the effects of maternal deficiency of folic acid during pregnancy on pulmonary development and protein A (SP-A) expression in newborn rats in order to explore the possible mechanism of lung developmental disorders.</p><p><b>METHODS</b>Thirty-six adult Sprague-Dawley female rats were randomly assigned into two groups: control and study (n=18). The study and the control groups were fed with fodder containing folic acid or not respectively. Two weeks later, the female rats in the two groups copulated with normal male rats. Newborn rats were sacrificed at 1, 7 and 14 days after birth (8 pups at each time point). Lung sections were stained with hematoxylin and eosin for histological examination. SP-A expression of protein and mRNA were determined by immunohistochemistry and real-time quantitative RT-PCR, respectively.</p><p><b>RESULTS</b>The newborn rats from the study group showed damaged lung tissue structures. The mean optical density of type II cells with positive expression of SP-A decreased significantly from 1 to 14 days in newborn rats of the study group compared with the control newborn rats (P<0.05). The real-time quantitative RT-PCR showed that the expression of lung SP-A mRNA also decreased significantly from 1 to 14 days in newborn rats of the study group compared with control newborn rats (P<0.05).</p><p><b>CONCLUSIONS</b>Maternal deficiency of folic acid during pregnancy can decrease the expression of SP-A in lung tissues of newborn rats, which might lead to the disorder of lung development maturation.</p>

Distribution and expression of transforming growth factor beta and their receptors in hypertrophic scar / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the role of transforming growth factor beta (TGF beta) and their receptors (TGF beta-R) in the pathogenesis of hypertrophic scar.</p><p><b>METHODS</b>Specimens of normal skin and hypertrophic scar were harvested and the distribution and the expression of the TGF beta and TGF beta-R were determined by immunohistochemistry and in situ hybridization method.</p><p><b>RESULTS</b>The expressions of TGF beta and TGF beta-RII in normal skin were higher than the expression of TGF beta 1, TGF beta 2 and TGF-RI. But in hypertrophic scar the results were on the contrary. The mRNA expressions of TGF beta 1, TGF beta 2 and TGFRI were evidently increased with decreased mRNA expression of TGF-beta 3 and TGFR II in the hypertrophic scar when compared with those in the normal skin.</p><p><b>CONCLUSION</b>The expression of TGF-beta (beta 1, beta 2, beta 3) and their receptors in different levels during the process of wound healing might be related to the formation of hypertrophic scars.</p>

Experimental study of human skin fibroblasts cultured in three-dimension(3D) / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL).</p><p><b>METHODS</b>The human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morphology.</p><p><b>RESULTS</b>The flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles.</p><p><b>CONCLUSIONS</b>The proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better.</p>

Clinical observation of the long-term effects of rhEGF on deep partial-thickness burn wounds / 中华烧伤杂志

<p><b>OBJECTIVE</b>To evaluate the safety and long-term effect of recombinant human epithelial growth factor (rhEGF) on deep partial-thickness burn wounds.</p><p><b>METHODS</b>Thirty-seven burn patients were enrolled in this study and were observed by randomized, double-blinded and placebo-controlled protocol. An area of deep partial-thickness burn wounds from each patient was divided into control (C) and treatment (T) portions. The wound in C was treated with normal saline while that in T with rhEGF. The patients were followed-up for 1 and 4 years after wound healing. The healed wounds were evaluated by modified Vancouver scar scale in terms of scar index (SI).</p><p><b>RESULTS</b>1 year after wound healing, it was found that the SI in T group (7.19 +/- 1.67) was obviously lower than that in C group (8.92 +/- 1.78, P < 0.01). The SI in T group (6.12 +/- 1.54) was still evidently lower than that in C group (8.09 +/- 1.81, P < 0.01) four years after wound healing. There were no signs of development of tumor or cancer in all the tested burn wound areas.</p><p><b>CONCLUSION</b>External application of rhEGF might be beneficial to the healing quality of deep partial-thickness burn wound with less scar formation and better long-term effects, and it is safe.</p>

Clinical study on the comparison of prophylactic tracheotomy with emergent tracheotomy after inhalation injury / 中华烧伤杂志

<p><b>OBJECTIVE</b>To compare prophylactic with emergent tracheostomy in 93 patients with inhalation injury, so as to explore the clinical significance of prophylactic tracheostomy.</p><p><b>METHODS</b>Tracheostomy was applied in all patients with moderate and severe inhalation injury. The patients with obvious dyspnea, low oxygen pressure and decreased oxygen saturation before the operation were set to be emergent tracheostomy group (ET, n = 21), while those without dysfunction of ventilation and gas exchange, as prophylactic group (PT, n = 72). The average operation starting time was 4.31 +/- 3.04 PBH and 34.47 +/- 2.79 PBH in PT and ET groups, respectively.</p><p><b>RESULTS</b>There were evident decrease of blood oxygen pressure and oxygen saturation and increase of respiratory and heart rates before the tracheostomy in ET group, but all these signs improved after the operation. In contrary, the vital signs in PT group exhibited stable perioperatively. Assisted respiration with respirator was applied in 20 cases (95.24%) in ET and in 65 cases (90.28%) in PT groups within 2 post-operative days (POD).</p><p><b>CONCLUSION</b>Early and prophylactic tracheostomy should be advocated in patients suspected to be with moderate or severe inhalation injury. Emergent tracheostomy must be avoided and respiratory assistance should be employed as early as possible.</p>

An observation of the effects of recombinant human transforming growth factor beta3 on fibroblast / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the role of recombinant human transforming growth factor beta3 (rhTGFbeta3) on fibroblast and its possible mechanism.</p><p><b>METHODS</b>Normal skin fibroblast (NSFb) and hypertrophic scar fibroblast (HSFb) were cultured in vitro, and were processed by different concentrations of rhTGFbeta3. NSFb and HSFb in DMEM solution without rhTGFbeta3 were employed as control. The changes in the protein and mRNA expression of type I and III collagen in NSFb and HSFb were observed.</p><p><b>RESULTS</b>(1) The expression of type I and III procollagen in NSFb was evidently different from that of HSFb (2) The synthesis of type I and III procollagen in all test groups was increased obviously after rhTGFbeta3 process (P < 0.001) while the ratio of type I to III procollagen was decreased when compared with that in control group. (3) The effects of rhTGFbeta3 on the biological behavior exhibited an obvious dose- effects relationship. The contents of type I to III procollagen in HSFb were higher than those in NSFb when the dose of rhTGFbeta3 was same.</p><p><b>CONCLUSION</b>rhTGFbeta3 could effectively promote the synthesis of type I and III procollagen, especially type III procollagen in fibroblasts. This might be beneficial to the accelerate of wound healing and to inhibit or prevent scar formation.</p>