AIM:To obtain the fine safflower essential oil by supercritical CO_2 extraction(SFE-CO_2). METHODS: The effects of pressure, temperature and CO_2 flow rate on supercritical CO_2 extraction were studied and the safflower essential oil was analysed by GC-MS. Both yield and components of essential oil obtained by SFE-CO_2 were compared with that obtained from hydrodistillation and microwave-assisted extraction. RESULTS: The optimal conditions were determined: the pressure of 9 MPa, the temperature was at 40 ℃ and the CO_2 flow rate was 4 kg/h. The yield of essential oil by SFE-CO_2 was higher than that of hydrodistillation, the qualities of both kinds of essential oil were fine, as analysed by GC-MS. The yield of essential oil by microwave extraction was the highest, but its quality was worse than those obtained by using other methods. CONCLUSION: The SFE-CO_2 is a good method for extraction of safflower essential oils.

Objective:To analyze the association between CRHR1 gene(rs1876828)polymorphism and the effect of inhaled corticosteroids(ICS)in children with bronchial asthma.Methods:A total of 60 children with moderate persistent asthma who were treated in the Department of Pediatrics of the First Affiliated Hospital of Harbin Medical University from January 2018 to June 2019 were included.The CRHR1 gene rs1876828 locus in children with asthma was detected by Sanger sequencing.The children were divided into TT genotype group(TT group) and CC genotype group(CC group)according to the different base sequences of gene loci.There were 22 cases in TT group(36.7%)and 38 cases in CC group(63.3%). Both groups were given aerosol inhalation of ICS and symptomatic treatment.Clinical symptoms and signs were observed and scored before and after treatment for 3d, 10d and 30d, and the days required for complete disappearance of symptoms and signs were recorded.Results:After 3d of treatment, clinical symptoms and signs of TT group and CC group were improved to a certain extent, but there was no statistical significant difference between two groups( P>0.05). At 10d and 30 d after treatment, the recovery of the two groups was better than that at 3d, and the improvement degree of the TT group was significantly better than that of the CC group, with statistical significance( P<0.05). The time of complete remission of symptoms and signs in TT group and CC group was(8.68±7.42)d and(16.21±7.82)d; the difference was statistically significant( P<0.01). Conclusion:There is a polymorphism of CRHR1 rs1876828 locus in children with bronchial asthma, which manifests as TT genotype and CC genotype, and CC genotype is the majority.The polymorphism of CRHR1 gene rs1876828 in asthmatic children is associated with the efficacy of ICS.The efficacy of ICS in children with TT genotype is better than that of CC genotype.

Objective To investigate de-escalation of empiric broad-spectrum antibiotics treatment for patients in intensive care unit (ICU).Methods Data of the patients discharged from ICU in the Second Affiliated Hospital of Zhejiang University from July 1 to December 31 of 2012 and from July 1 to December 31 of 2013 were retrospectively reviewed.Patients with initial use of empirical broad-spectrum antibiotics within 3 d after ICU admission were included in the study.Clinical data including status of infection,the initial empiric antimicrobial therapy,pathogens culture and adjustment of antibiotics in 5 days were analyzed.Results A total of 841 patients were discharged from ICU during the study periods and antibiotics were used in 786 (93.5％) patients.Among 786 patients,389 (49.5％) were treated empirically with broad-spectrum antibiotics,but only 269 (69.2％) had evidences of bacterial infections.Of the 389 patients with empiric antibiotics use,de-escalation of antibiotics was applied only in 6 (1.54％) patients within 5 days after the initiation of treatment.In 269 patients with evidence of infection,specimen sampling and culture were performed in 248 (92.2％) patients within 3 days,among which 165 samples were positive,and the clinical isolates were mainly multi-drug resistant gram negative bacilli and colonized bacteria in oropharyngeal cavity.De-escalation was applied only in 4 (1.49％,4/269) patients with evidences of bacterial infections.Conclusion Broad-spectrum antibiotics as initial empiric therapy is common for patients in ICU,however de-escalation of empiric therapy is rarely applied even in patients with positive results in pathogen isolation and culture.

Objective:To study the inhibitory effect and anti-cancer mechanisms of adenovirus-mediated ING4 gene on the MG-63 osteosarcoma xenografts in nude mice.Methods:Ad-ING4 was transfected into QBI-293 cells and harvested.15 nude mice of the subcutaneous tumor models were established with MG-63 osteosarcoma cells and were randomly divided into PBS,Ad-GFP and Ad-ING4 groups.Then PBS(100 μl),Ad-GFP(100 μl,10~9pfu/ml) and Ad-ING4 (100 μl,10~9pfu/ml) for each one were given respectively QOD for 5 times,with intratumor injections.Tumor volume changes were monitored;and the 15 mice were sacrificed 2 weeks after treatment,the tumors were removed,weighed and ratios of tumor-suppression were calculated.The morphological changes of apoptotic tumor cells were observed under microscope.Bcl-2,Bax,Caspase-3,VEGF,CD34 expression was tested by immumohistochemistry.Results:High titer(10~9pfu/ml)adenoviral vector of ING4 gene were obtained.In nude mice bearing MG-63 osteosarcoma xenografts,the growth of MG-63 tumors treated by intratumoral injecting of Ad-ING4 was significantly suppressed,compared with PBS group and Ad-GFP group.The ratios of tumor weight-suppression of Ad-ING4 group was 59.3%(P＜0.05).Immumohistochemistry displayed that the expression of Bax,Caspase-3 was up-regulated and the expression of Bcl-2,VEGF,CD34 was down-regulated by Ad-ING4.Conclusion:Ad-ING4 can inhibit the growth of MG-63 osteosarcoma xenografts in nude mice,which may be via activating the apoptosis pathway and inhibiting tumor angiogenesis.

Objective To establish Ad-hLIF transgenic feeder cells for the expansion of umbilical cord blood CD34+ HSPC in vitro and study the SCID mice model of hematopoietic stem/progenitor cell (HSPC) transplantation. Methods The expression of objective gene in Ad-hLIF transgenic feeder cells was detected by RT-PCR and ELISA. The purity of umbilical cord blood CD34+ HSPC separated by magnetic-activated cell sorting(MACS) was detected by the flow cytometry. After expanded with various combinant of cytokines and transgenic feeder layer cells for 28 d, the quantity of mono-nuclear cell (MNC) and CD34+ cells rate was detected in different time. MNC after expansion stained by CFDA SE was injected to the sublethally irradiated SCID mice. Humanize gene Alu was detected by RT-PCR and fluorescence microscope. Results The green fluorescence was observed in the transgenic cells infected with 50MOI( multiplicity of infection) Ad-hLIF, and the objective gene was confirmed by RT-PCR and ELISA. The purity of umbilical cord blood CD34+ HSPC separated by MACS could reach 95.60% ±2.58%, Ad-hLIF transgenic feeder cells and various cytokines system increased MNC by 356.95±0.87 fold, and maximal expansion of CD34+ cells was observed during 0-14 d of culture, then down-expansion gradually. Four weeks after transplanted in SCID mice, fluorescently-labeled humanize cells still can be observed. The existence of the humanized gene Alu was confirmed by RT-PCR. Conclusion Ad-hLIF transgenic feeder cells can effectively proliferate umbilical cord blood CD34+ HSPC in vitro and delay it differentiate, what's more, it has high transplant efficacy and haematogenesis activity.

Objective:To investigate the association between gene polymorphisms in vitamin D receptor(VDR) and Tourette syndrome (TS).Methods:The genetic contributions of VDR FokI (rs2228570), BsmI (rs1544410), and Cdx2 (rs11568820) polymorphisms were genotyped by TaqMan allelic discrimination real-time (RT)-PCR, which evaluated by a case-control analysis in 417 TS patients and 442 healthy controls, and followed by a family-based study in 417 TS trios.Chi-square test and relative risk analysis were conducted by IBM SPSS 23.0 software.Results:FokI (rs2228570) had three genotypes(CC=109, CT=235, TT=73); BsmI (rs1544410) had three genotypes(AA=2, AG=45, GG=370); Cdx2 (rs11568820) had three genotypes(AA=71, AG=200, GG=146). No significant difference in genotype ( χ2=5.516, P=0.063; χ2=3.466, P=0.177; χ2=0.561, P=0.755, respectively) or allele frequencies( χ2=0.840, P=0.359; χ2=3.376, P=0.066; χ2=0.051, P=0.822, respectively)of FokI, BsmI and Cdx2 were identified between TS patients and control groups.No significant over-transmission was identified for these three polymorphisms among 417 TS trios in the family-based study (TDT for FokI: χ2=0.009, P=0.962; for BsmI: χ2=1.220, P=0.320; and for Cdx2: χ2=0.260, P=0.646). Haplotype relative risk (HRR) analysis and haplotype-based haplotype relative risk (HHRR) analysis showed no significant difference in allele frequencies distribution of FokI, BsmI and Cdx2 (all P>0.05). Conclusion:VDR receptor gene polymorphism has no effect on TS susceptibility in the Chinese Han population. However, a potential role of VDR should be explored in more polymorphisms, different populations and larger samples.

Objective:To analyze the clinical phenotype and genetic etiology of a child with Multiple congenital anomalies-hypotonia-seizures syndrome 1 (MCAHS1).Methods:Clinical data of a 2-year-old boy who had presented at the Affiliated Hospital of Qingdao University in March 2023 for "intermittent limb twitching for 2 years" was collected. Peripheral blood samples were collected from the child and his parents for whole-exome sequencing (WES). Candidate variants were verified by Sanger sequencing and bioinformatic analysis based on the guidelines from the American College of Medical Genetics and Genomics (ACMG).Results:The child had manifested with distinctive facial features, limb deformities, hypotonia, motor and intellectual delays, and epileptic seizures. WES revealed that he has harbored compound heterozygous variants of the PIGN gene, namely c. 963G>A (p.Q321=) and c. 994A>T (p.I332F), which were inherited from his phenotypically normal mother and father, respectively. Based on the ACMG guidelines, the c. 963G>A was classified as a pathogenic variant (PVS1+ PM2_Supporting+ PM3), whilst the c. 994A>T was classified as a variant of uncertain significance (PM2_Supporting+ PP3). Conclusion:Above discovery has expanded the mutational spectrum of the PIGN gene variants associated with MCAHS1, which may facilitate delineation of its genotype-phenotype correlation.

Objective To observe the clinical efficacy and safety of GINA regimen and GINA regimen combined with oral Huaiqihuang granule in the treatment of children with bronchial asthma.Methods A ran-domized,single blind,multicenter,parallel controlled clinical trial wascarried out.A total of 1 128 patients with bronchial asthma in children were randomized into two groups.The observation group were treated with GINA regimen combined with oral Huaiqihuang granule.The GINA regimen treatment group was treated by GINA reg-imen.Clinical assessment and C-ACT scores was observed in first month,third month,sixth month after treat-ment.Clinical assessment included the times of upper respiratory tract infection occurrence,bronchitis and pneu-monia,asthmatic attacks,application of emergency medicine,hospitalizations due to asthmatic.Drug adverse effect in the two groups was compared.Results The times of upper respiratory tract infection,bronchitis and pneumonia,asthmatic was significantly decreased(P <0.05 ),and C-ACTscores were significantly higher(P <0.05)in the first month,the third month,and the sixth month after treatment.There were 16 cases of drug relat-ed adverse reactions.Seven cases were in observation group(n ﹦7)(1.15%)and 9 cases in the GINA regimen treatment group(n ﹦9)(1.73%).There was no significant difference of adverse effect between the two groups (P >0.05).Conclusion The treatment of bronchial asthma in children with GINA regimen combined with oral Huaiqihuang granule can significantly reduce the incidence of respiratory infections and the number of asthmatic attacks dramatically and safely improve clinical curative effect,asthma control.

To study the inhibitory effect of a recombinant adenoviral vector carrying human IL-24 gene on SGC-7901 human gastric cancer cell. We infected the SGC-7901 gastric cancer cells with Ad blank adenovirus at various multiplicity of infection (MOIs) to find the optimal infective dose. The SGC-7901 tumor cells were infected with Ad-IL-24 at the optimal MOI in the following experiments. Adenovirus-mediated IL-24 transcription expression in SGC-7901 cells was examined by RT-PCR. The growth-suppressing effect of Ad-IL-24 on SGC-7901 tumor cells was assessed by MTT assay. Apoptosis and cell cycle of SGC-7901 tumor cells infected with Ad-IL-24 was evaluated by flow cytometer (FCM), respectively. The karyomorphology of apoptotic SGC-7901 tumor cells was examined using Hoechst33258 staining under fluorescence microscopy. The expression of apoptosis-related genes was future determined by semi-quantification RT-PCR; We demonstrated that the MOI of 100 was the optimal infective dose in the study on adenovirus-mediated IL-24 gene transfer into SGC-7901 gastric cancer cell; IL-24 gene mediated by adenovirus could successfully transcribe in SGC-7901 tumor cells; Ad-IL-24 could significantly inhibit SGC-7901 tumor cell growth and induce apoptosis, it also can up-regulate the express of bax, caspase-3 and p53 whilst down-regulate the bcl-2 expression. Thus, adenovirus-mediated IL-24 expression had marked anti-tumor effect in suppressing SGC-7901 human gastric cancer cell growth and inducing apoptosis, which may be closely associated with its up-regulation of bax/bcl-2, caspase-3 and p53.
Adenoviridae ; genetics ; metabolism ; Apoptosis ; genetics ; Caspase 3 ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Interleukins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Up-Regulation ; bcl-2-Associated X Protein ; genetics ; metabolism