OBJECTIVE: To study the percentages of polymorphonuclear leukocytes (PMN), mononuclear cells (MNC) and fibroblastic cells (FBC) in different post-traumatic intervals after skeletal muscle mechanical injury in rats. METHODS: The rat model of skeletal muscle mechanical injury was established. The rats were divided into injured groups (6 h, 12 h, 1 d, 3 d, 7 d, 10 d and 14 d after injury) and control group. The percentages of PMN, MNC and FBC in different post-traumatic intervals after skeletal muscle mechanical injury were assessed with HE staining and image analysis. RESULTS: At post-injury 6-12h, the percentages of PMN and MNC infiltration appeared in injured sites and that of PMN reached peak. At 1 d, the percentage of MNC infiltration appeared and reached peak, while that of PMN decreased. At 3-7 d, the percentage of FBC gradually increased, while that of PMN and MNC decreased. At 10-14d, the percentage of FBC reached peak. CONCLUSION The percentages of PMN, MNC and FBC in injured zones showed time-dependent changes, which might be used as reference index for determination of age of skeletal muscle injury.
Animals ; Fibroblasts ; Muscle, Skeletal/injuries* ; Neutrophils ; Rats ; Time Factors

Chuanniuxi (Radix Cyathule) is one of the most important geo-herb in Sichuan province, which adulterants are Hongniuxi (Cyathula capitata) and Huainiuxi (Achyranthes bidentata). In this paper Chuanniuxi and its adulterants were identified by SCAR markers. Nineteen populations from Tianquan, Baoxin, Huili and Jinkouhe were collected and their RAPD fingerprints were established. Based on the RAPD patterns, two polymorphic bands F300 and F500 were selected, recycled, cloned and sequenced. According to the sequences two pairs of sequence characterized amplified regions (SCAR) primers were designed and used to amplify all materials to prove the efficiency of identification of the different populations. Chuanniuxi and Huiniuxi could be distinguished by the primer SC-320, Chuanniuxi and Hongniuxi could be distinguished by the primer SC-495. Combining the two SCAR markers, Chuanniuxi, Hongniuxi and huainiuxi could be identified effectively and quickly.
Achyranthes ; genetics ; Amaranthaceae ; genetics ; Cloning, Molecular ; DNA, Plant ; genetics ; Drug Contamination ; Genetic Markers ; genetics ; Nucleic Acid Amplification Techniques ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; Sequence Analysis, DNA ; Time Factors

Objective:To evaluate whether midazolam with propofol target controlled infusion (TCI intravenous sedation during the mandibular third molar extraction influences patients' perioperative anxiety.Methods:The subjects were patients who planned to undergo the mandibular third molar extraction in Peking University School and Hospital of Stomatology,whose state anxiety inventory (SAI) scores were ≥ 38 at the initial visit.They were divided into intravenous sedation group (IVS) and local anesthesia group (LA) on the basis of the planned intravenous sedation.Each group was divided into two subgroups according to the overall SAI scores at the initial visit:IVS-Ⅰ,LA-Ⅰ (SAI:38-50) and IVS-Ⅱ,LAⅡ (SAI:51-80).The anxiety before and after the surgery was evaluated by the SAI scores at the initial visit (T1),before surgery (T2) and 7 days after surgery (T3).The anxiety during the surgery was evaluated by the heart rate,blood pressure and visual analogue scale (VAS) scores.Results:There were no significant differences on SAI at T1,T2,and T3 in the two groups (P ＞ 0.05).The heart rate,blood pressure and VAS pain scores of IVS group were significantly lower than those of LA group during the surgery (P ＜ 0.001).Conclusion:Intravenous sedation with midazolam and propofol TCI was effective on the patients' anxiety during the third molar extraction,which successfully made the patients more comfortable and their heart rate,blood pressure and oxygen saturation more stable during the surgery.But there were no significant differences on the patients' anxiety at the initial visit (T1),before surgery (T2) and 7 days after surgery (T3) according to the SAI scores in the two groups.

OBJECTIVE: To research the relation between the time-dependent appearances of myotibroblasts during the repair of contused skeletal muscle in rat and wound age determination. METHODS: A total of 35 SD male rats were divided into the control and six injured groups according to wound age as follows: 12 h, 1 d, 5 d, 7 d, 10 d and 14 d after injury. The appearances of myofibroblasts were detected by HE staining, immunohistochemistry and confocal laser scanning microscopy. Masson's trichrome staining was utilized to examine collagen accumulation in the contused areas. RESULTS: Immunohistochemical staining showed that α-SMA+ myofibroblasts were initially observed at 5 d post-injury. The average ratio of myofibroblasts was highest at 14 d post-injury, with all samples, ratios more than 50%. In the other five groups, the average of α-SMA positive ratios were less than 50%. The collagen stained areas in the contused zones, concomitant with myofibroblast appearance, were increasingly augmented along with advances of posttraumatic interval. CONCLUSION The immunohistochemical detection of myofibroblasts can be applied to wound age determination. The myofibroblasts might be involved in collagen deposition during the repair of contused skeletal muscle in rat.
Animals ; Collagen/metabolism* ; Contusions/metabolism* ; Immunohistochemistry ; Male ; Microscopy, Confocal ; Muscle, Skeletal/metabolism* ; Myofibroblasts/metabolism* ; Rats ; Time Factors ; Wound Healing

In the treatment of hypertensive crisis, the novel Rho kinase inhibitor DL0805-2 can rapidly lower systematic blood pressure, reduce pulmonary artery pressure, and has a significant protective effect on lung injury. This experiment intends to evaluate the efficacy of DL0805-2 against pulmonary arterial hypertension (PAH) and preliminarily reveals its underlying mechanism. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences. Sprague Dawley (SD) rats were randomly divided into DL0805-2 low, medium, and high dose groups (1, 3, and 10 mg·kg^-1), bosentan positive control group, model group, and blank control group. The drug was administered daily on the 7th day after model establishment by monocrotaline injection. On the 25th day of the experiment, relevant indicators were examined to observe the therapeutic effect of DL0805-2 on pulmonary hypertension. DL0805-2 significantly relieved the abnormal changes in the physiological parameters related to PAH induced by monocrotaline, including reducing right ventricular systolic pressure, alleviating cardiac damage caused by pressure overload, and reducing the levels of endothelin-1 and inflammatory factors in lung tissues. DL0805-2 also attenuated pulmonary arteries remodeling. It was preliminarily discovered that DL0805-2 exerts preventive and therapeutic effect on PAH through Rho-kinase pathway. Our results suggested that DL0805-2 had good therapeutic effects on monocrotaline-induced PAH rat model. It intervened early in the disease process, effectively prevented the development of the disease, and reduced the mortality of the diseased animals. The mechanism is related to Rho-kinase pathway.

OBJECTIVERespiratory syncytial virus (RSV) infects nearly all children under two years of age. It is poorly understood why a few children who were infected with RSV develop bronchiolitis that require hospital admission while most have a relatively minor illness. Several recent studies have obtained some indications for the involvement of genetic heterogeneity in RSV bronchiolitis, implying that the clinical outcome of RSV infection perhaps is determined by genetic factors. Regulated on activation, normal T cell expressed and secreted RANTES plays a key role in the pathophysiology of RSV bronchiolitis. The purpose of this study was to explore the genetic association between the RANTES gene promoter -28C/G polymorphism and RSV bronchiolitis in Chinese Han ethnic group population.

METHODSThe study recruited 238 hospitalized patients (186 male and 52 female) under 12 months of age, with a clinical diagnosis of bronchiolitis due to RSV, the sex, age, hospital stay, SaO2 at the time of admission, personal and family history of atopy were recorded. The 288 healthy control subjects (206 male and 82 female), who had no evidence of personal or familial history of atopy and no history of wheezing, were chosen at the same time. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to identify the polymorphism at position -28C/G of the RANTES promoter. The total IgE concentrations in serum samples were measured by enzyme-linked immunosorbent assay (ELISA). The absolute peripheral blood eosinophil counts were measured by using an automated hematology analyzer.

RESULTSThe distribution of RANTES -28C/G gene polymorphism was in accordance with Hardy-Weinberg equilibrium. Compared to control subjects, significant difference was demonstrated for genotypes and allele frequencies of the RANTES -28C/G polymorphism in patients with RSV bronchiolitis (G = 10.22, P < 0.01; chi2 = 9.708, P < 0.01). Compared with the wild type CC, the -28G allele carriers demonstrated a 2.09-fold increased risk of RSV bronchiolitis (OR = 2.09, 95% CI = 1.32 - 3.30, P < 0.01). Interestingly, both the percentage of personal history of atopy and the percentage of family history of atopy for the -28G allele carriers were significantly higher (P < 0.05) than that for those CC homozygotes carriers in RSV bronchiolitis. Compared with the wild type CC, the -28G allele carriers demonstrated a 1.85-fold increased risk of the personal history of atopy (OR = 1.85, 95% CI = 1.01 - 3.38, P = 0.045) and a 1.91-fold increased risk of the family history of atopy (OR = 1.91, 95% CI = 1.03 - 3.54, P = 0.037), and the absolute peripheral blood eosinophil counts for the -28G allele carriers were significantly higher (P < 0.05).

CONCLUSIONThe RANTES gene promoter -28C/G polymorphism is associated with the susceptibility to RSV bronchiolitis, and the -28G allele is an important predisposing factor for the personal history of atopy and the family history of atopy in RSV bronchiolitis.

Alleles ; Bronchiolitis ; genetics ; virology ; Case-Control Studies ; Chemokine CCL5 ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Humans ; Infant ; Male ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Respiratory Syncytial Virus Infections ; complications ; genetics

OBJECTIVERespiratory syncytial virus (RSV) infects nearly all children under two years of age. It is not understood why some develop serious bronchiolitis. Whether there is a genetic component is not known. The nature of the association between RSV bronchiolitis and subsequent wheezing remains unknown. interleukin-8 (IL-8) is a potent neutrophil chemokine and activator, which plays a role in virus-induced wheezing diseases. The purpose of this study was to assess the genetic association between the IL-8 gene promoter -251A/T polymorphism and RSV bronchiolitis and post-bronchiolitis wheezing in children.

METHODSTotally 320 children who were hospitalized for bronchiolitis together with positive immunofluorescence for RSV were recruited in this study from Jan. 2002 to Jan. 2004. A group of 272 healthy children were enrolled as controls. The age of these children ranged from 1 to 12 months. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to identify the polymorphism at position-251 of the IL-8 promoter in RSV bronchiolitis and control groups. The total IL-8 and IgE concentrations in serum samples were measured by enzyme-linked immunosorbent assay (ELISA). The patients with RSV bronchiolitis were followed up in order to analyze the occurrence of wheezing post-bronchiolitis.

RESULTS(1) Both A allele and T allele were detected at -251 of the IL-8 promoter; the prevalence of the A allele in RSV bronchiolitis group was 45.6%, as compared with 37.7% in normal group. The prevalence of IL-8-251A allele was significantly different between the two groups (P < 0.05). (2) For genotypes T/T, A/T, A/A in RSV bronchiolitis, level of serum IL-8 were (17 +/- 6) ng/L, (21 +/- 7) ng/L, (24 +/- 9) ng/L, respectively, the difference was significant among the three genotypes (P < 0.01). (3) The prevalence of the A allele in the group who wheezed after the episode of RSV bronchiolitis was 54.6%, as compared with 35.8% in the group who had bronchiolitis but did not go on to wheeze. The prevalence of IL-8-251A allele was significantly different between the two groups (P < 0.05).

CONCLUSIONPolymorphism of IL-8 promoter-251A/T was associated with susceptibility to RSV bronchiolitis in children. The association of IL-8-251A with severe RSV bronchiolitis is most marked in the children who go on to wheeze.

Adolescent ; Alleles ; Bronchiolitis ; complications ; Child ; Child, Preschool ; Chromosome Mapping ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Interleukin-8 ; genetics ; Male ; Polymorphism, Genetic ; Respiratory Sounds ; etiology ; genetics ; Respiratory Syncytial Virus Infections ; complications ; virology

OBJECTIVETo observe the correlation factor about early-life RSV bronchiolitis and sequential recurrent wheezing for two years.

METHODSFollow up the RSV bronchiolitis patients for two years in order to analyze the occurrence of wheezing post-bronchiolitis. Single and multiple logistic regression analysis were used to determined the risk factors such as individual atopy history and familial atopy history, pet feeding, breast milk, secondhand smoke for RSV bronchiolitis and subsequent wheezing.

RESULTS(1) Not breast feeding, exposure to cigarette smoke and the deficiency of VitA, D were the significant risk factors contributed to the RSV branchiolitis. (2) Exposure to cigarette smoke, the deficiency of VitA, D, the personal history of atopy and the family history of atopy were the significant risk factors contributed to the post-bronchiolitis wheezing in children. (3) Those patients who eosinophilia, high serum IgE, RANTES and decreased TH1 to TH2 Ratio were more likely to have wheezing after RSV bronchiolitis.

CONCLUSION(1) Not breast feeding, exposure to cigarette smoke and the deficiency of VitA, D were the significant risk factors contributed to the RSV bronchiolitis. (2) Exposure to cigarette smoke, the deficiency of VitA, D, the personal history of atopy and the family history of atopy were the significant risk factors contributed to the post-bronchiolitis wheezing in children.

Breast Feeding ; Bronchiolitis ; complications ; immunology ; metabolism ; virology ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Respiratory Sounds ; etiology ; immunology ; Respiratory Syncytial Virus Infections ; complications ; immunology ; metabolism ; virology ; Respiratory Syncytial Virus, Human ; physiology ; Risk Factors ; Tobacco Smoke Pollution ; adverse effects ; Vitamin A ; metabolism ; Vitamin D ; metabolism

The therapeutic potential of curcumin (Cur) is hampered by its poor aqueous solubility and low bioavailability.The aim of this study was to determine whether Cur nanoemulsions enhance the efficacy of Cur against prostate cancer cells and increase the oral absorption of Cur.Cur nanoemulsions were developed using the self-microemulsifying method and characterized by their morphology,droplet size and zeta potential.The results showed that the cytotoxicity and cell uptake were considerably increased with Cur nanoemulsions compared to free Cur.Cur nanoemulsions exhibited a significantly prolonged biological activity and demonstrated better therapeutic efficacy than free Cur,as assessed by apoptosis and cell cycle studies.In siru single-pass perfusion studies demonstrated higher effective permeability coefficient and absorption rate constant for Cur nanoemulsions than for free Cur.Our study suggested that Cur nanoemulsions can be used as an effective drug delivery system to enhance the anticancer effect and oral bioavailability of Cur.

OBJECTIVETo investigate the relationship between the distribution of mononucleotide polymorphism of five regulation regions of alpha-IFN among HLA-DRB1*11 gene episodes and the therapeutic effects of alpha-IFN treatment in chronic hepatitis B patients.

METHODSOne hundred seven chronic hepatitis patients from Nanjing Second Hospital who were treated by alpha-IFN for 12 months and then followed at least six months without the treatment were randomly selected for this regressive analysis. They were grouped into a continuous responsive group and a non-continuous responsive group. Hepatitis B virus X interacting protein gene locus was searched in NCBI. Single nucleotide polymorphism (SNP) gene locus was detected based on a pooling sequencing method. Primer and TaqMan-MGB probes referring to different mononucleotide loci were designed respectively to detect SNP in five regulation regions of alpha-IFN. Then gene sequencing differences between the two groups were analyzed.

RESULTSAmong the 107 cases there were 30 cases (28.0%) in the continuous responsive group and 77 cases (71.9%) in the non-continuous responsive group. CT occupation rate in five regulation regions of IFN reached 18.0% in the continuous responsive group and 23.8% in the non-continuous responsive group. AG occupation rate reached 10.8% in the former group and 15.8% in the latter group. The differences in CT and AG between the two groups were significant.

CONCLUSIONSThe distribution of mononucleotide polymorphism of five regulation regions of alpha-IFN among HLA-DRB1*11 gene episodes affects the IFN anti-virus treatment. Detecting the gene distribution of mononucleotide in five regulation regions of alpha-IFN helps in predicting the therapeutic effects of alpha-IFN.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; Genotype ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; genetics ; Humans ; Interferon-alpha ; therapeutic use ; Polymorphism, Single Nucleotide ; Regression Analysis ; Treatment Outcome ; Young Adult