This study is aiming to establish an efficient metabolite extraction method for exploration of molecular mechanisms of desiccation tolerance of the resurrection plant Boea hygrometrica using a metabolomics approach. The extracts of metabolite in B. hygrometrica using methanol solution (method A) and methanol-chloroform-water solution (method B) were analyzed by gas chromatography-mass spectrometry (GC-MS). The total numbers of chromatographic peaks, extraction efficiency, retention time and the peak stability were compared. The results showed that for fresh materials, the total chromatographic peak number of method B is more than that of method A; the extraction efficiency of nine representative metabolites by method B is higher than that by method A; the comparison of 10 random chromatographic peaks revealed that the relative standard deviation (RSD) values of the retention time are less than 1% for both methods, whereas the RSD values of the extraction efficiency is different. The percentage of peaks that owned RSD values of the extraction efficiency higher than 10% is 50% for method A and 100% for method B. In addition, method B was also efficient for dry materials from B. hygrometrica. The number of chromatographic peaks, RSD value of retention time and extraction efficiency of dry materials was similar to that of fresh materials using method B, but decreased sharply using method A. Putting together, our study provided evidence that method B is an efficient extraction method for further analysis of metabolites from this resurrection species.
Chemical Fractionation ; methods ; Chloroform ; Gas Chromatography-Mass Spectrometry ; Magnoliopsida ; chemistry ; Metabolome ; Metabolomics ; Methanol ; Plant Extracts ; chemistry ; Solvents

Objective To evaluate the impact of small interference RNA (siRNA)-induced silencing of adrenomedullin (ADM) gene on the growth of and apoptosis in a malignant melanoma cell line A375.Methods Three siRNAs targeting ADM gene were constructed and transfected into A375 cells.Real-time quantitative reverse transcription PCR (qRT-PCR) was performed to measure the expression of ADM to select the most efficient siRNA for the next experiment.A375 cells were classified into 3 groups to be transfected with the selected siRNA (test group),non-specific sequences (negative control group),or remain untransfected (blank control group).Then,an immunohistochemical SP method was used to determine the expression of ADM at 24hours,methyl thiazolyl tetrazolium (MTT) assay to detect the proliferation of A375 cells at 24 and 48 hours,flow cytometry to observe cell apoptosis at 48 hours.Results qRT-PCR showed that ADM-siRNA-2 was the most efficient.The expression of ADM was significantly lower in the test group than in the other two groups.The proliferation level (represented as the absorbance value at 490 nm) of A375 cells was 0.389 ± 0.0375 and 0.469 ± 0.0330 in the test group at 24 and 48 hours respectively,significantly lower than that in the negative control group (0.548 ± 0.0376,0.676 ± 0.0374,both P＜ 0.05) and blank control group (0.574 ± 0.0733,0.685 ± 0.0154,both P ＜ 0.05).Flow cytometry revealed a statistical increase in early apoptosis rate in the test group compared with the blank control group and negative control group ((20.200 ± 6.046)％ vs.(1.667 ± 0.340)％ and (4.587 ± 1.294)％,both P ＜ 0.05).No significant differences were observed in the proliferation level or early apoptosis rate between the negative control group and blank control group (both P ＜ 0.05).Conclusion The siRNA-induced silencing of ADM gene can inhibit the proliferation of,but promote the apoptosis in,A375 cells.

0.05).Conclusion The thin section collodion anatomy of temporal bone area,combined with the images of HRCT can clearly delineate the details of facial recess and its relative structures.

Objective To investigate the clinical efficacy of umbilical blood stem cell transplantation (UCBSCT) on the treatment of hepatitis B liver cirrhosis. Methods Forty-eight patients with hepatitis B liver cirrhosis were enrolled and divided into the treatment group and the control group. There were 25 patients in the treatment group , who received UCBSCT treatment based on conventional liver protection treatment and 23 patients in the control group , who received conventional liver protection treatment. The changes of liver function , coagulation function, clinical symptoms, signs and side effects were studied before the treatment and at 2, 4, 12 and 24 weeks post-treatment. Results The levels of albumin, cholinesterase, and prothrombin activity in the treatment group were higher than those before treatment and were higher than those in the control group. The parameters in the control group were not significantly changed before and after the treatment (P > 0.05). The levels of alanine aminotransferase,aspartate aminotransferase,total bilirubin in both two groups were not significantly changed before and after the treatment (P > 0.05). After 4-week treatment,the differences on improvement of appetite , lacking in strength , abdominal distension , ascites were statistically significant in the treatment group compared with the control group (P < 0.05). No adverse reactions were observed in all groups. Conclusion UCBSCT on the treatment of hepatitis B liver cirrhosis is safe and reliable.

Objective To investigate the limitation of quantitative measurement of cerebral blood flow (CBF) of normal white matter by using a single subtraction with thin-slice TI1 periodic saturation (Q2TIPS Ⅱ ) pulsed arterial spin labeling (PASL)technique. Methods Thirty-one patients with brain tumors were examined at 3.0 T MRI system . A second version of quantitative imaging of perfusion using a single subtraction with additional thin-section periodic saturation after inversion and a time delay (Q2TIPS) technique of pulsed arterial spin labeling in the multisection mode and T2* dynamic susceptibility-weighted contrast-enhanced (T2* DSC)MR imaging were both implemented. Cerebral blood flow map obtained from PASL and DSC were reviewed. The regions of interest( ROI )were placed in the region of normal white matter contralateral to the lesion in the proximal and distal slices. In regions of interest, the signal intensity (SI)was measured from the maps of cerebral blood flow map obtained from PASL and DSC. Pair-t test was performed to determine if there were significant signal differences between proximal and distal slices. Pearson linear correlation analysis of signal intensity was performed for values from the same slices of PASL-CBF and DSC-CBF maps. Results In the deep white matter of distal slice, PASL-CBF map showed perfusion deficit while DSC-CBF map showed low CBF in the corresponding brain area. With the increased inversion time,the PASL-CBF map showed obviously improved perfusion signal in deep white matter (but still some perfusion deficit)and slightly decreased perfusion signal in grey matter. The mean signal of normal white matter measured from distal slices of PASL-CBF maps was( -22.1 ±55.5) ml· 100 g-1 · min-1 while it was (89.5 ±45.5) ml. 100 g-1 · min-1 in proximal slices. There was a significant difference of signal intensity from PASL-CBF maps between distal and proximal slices ( t = - 9. 512, P ＜ 0. 01 ＝, while no difference of signal intensity between distal[ (62. 8 ± 29.9) ml · 100 g-1 · min-1] and proximal slices [(57. 1 ±29.6) ml · 100 g-1 · min-1 ]was obtained from DSC-CBF maps(t= -1.607,P＞0.05). There was no significant correlation between PASL-CBF and DSC-CBF in both distal ( r = 0. 093, P ＞ 0. 05 ) and proximal slices ( r = - 0. 234, P ＞ 0. 05). ConclusionsPASL has limitation in the accurate quantification of cerebral blood flow of normal white matter. The quantification of CBF was affected by the limitations of the technique itself and the different parameters chosen..

Objective To summarize the clinical experience in treatment of the Lisfranc joint injury with open reduction and internal fixation at early stage. Methods Twelve patients ( including ten males and two females at average age of 34 years) with early stage Lisfranc joint injury received open reduction and screw/wire fixation from 2005 to 2010. According to the Myerson classification, there were two patients with type A, eight with type B and two with type C. All the patients received open reduction and internal fixation with screw or Kirschner wire within 17 days after injury. The post-operative function was estimated by mid-foot scoring scale of AOFAS. X-ray and CT scan were used in radiography estimation. Results All the patients were followed up for average 33 months ( range, 6-60 months). The mean score of post-operative mid-foot scoring scale of AOFAS was 74.5 points ( range, 53-96 points), with excellent result in eight patients, good in two and fair in two. The anatomical reduction was observed in eight patients and all the patients obtained bony union according to the results of X-ray and CT scan.There was no any complication found.Conclusions Open reduction and internal fixation is a good choice for the treatment of Lisfranc joint injury at early stage. A preoperative comprehensive analysis combined with clinical X-ray and CT scan is necessary.

Objective This study aims to investigate the association of apoB/apoA-1 ratio with insulin resistance (IR) in patients with non-alcoholic fatty liver disease (NAFLD) . Methods A total of 224 patients with NAFLD and 166 healthy subjects were enrolled as NAFLD group and control group. Weight, height and blood pressure were recorded. Serum levels of fasting blood glucose (FPG), insulin (Fins), lipids, glycated hemoglobin (HbA1c) were measured. Homeostasis model assessment-insulin resistance (HOMA-IR) indices were calculated. Results Compared with control group, NAFLD group had higher apoB/apoA-1 ratio (0.76 ± 0.28 vs 0.61 ± 0.26) and HOMA-IR (2.43 ± 1.68 vs 1.86 ± 1.61) . Spearman correlation analysis showed that in NAFLD group, HOMA-IR positively correlated with age, body mass index (BMI), triglycerides (TG), low-density lipoprotein cholesterol (LDL-c), apoB/apoA-1 ratio (r =0.34, P < 0. 05) and HbA1c, and negatively correlated with high-density lipoprotein cholesterol (HDL-c) . Multiple linear regression analysis revealed that apoB/apoA-1 ratio was still associated with HOMA-IR in NAFLD group after adjustment for age and BMI. Conclusion The apoB/apoA-1 ratio is closely associated with IR in patients with NAFLD. ApoB/apoA-1 ratio may play a role in the development of IR in NAFLD.

Quorum sensing is an important gene regulatory mechanism in Pseudomonas aeruginosa and controls the expression of numerous virulence factors. We designed and constructed a screening system for quorum-sensing inhibitors. We developed the system by using the lasI and rhlA promoters fused with promoterless sacB as reporters. Using this system we screened a number of Chinese herb extracts, and identified three herb extracts containing inhibitors to the quorum-sensing system and to its regulated genes. The screening system developed was highly efficient and sensitive. It could serve as a useful tool to identify herb compounds that block infections but unlikely render antibiotic resistance in pathogens.
Acanthaceae ; chemistry ; Anti-Bacterial Agents ; pharmacology ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Promoter Regions, Genetic ; Pseudomonas aeruginosa ; drug effects ; genetics ; Quorum Sensing ; drug effects ; Valerianaceae ; chemistry

Objective:To study the difference between BRCA gene mutations in hereditary breast and ovarian cancer syndrome (HBOC) and in sporadic ovarian cancer (SOC).Methods:This study was for exploratory research, the inclusion criteria were 284 patients with ovarian cancer admitted at Shanxi Provincial Cancer Hospital from November 2018 to December 2019, with high-throughput DNA sequencing including the full coding regions and exon-intron link regions of BRCA1 and BRCA2 gene. Pathogenic mutations in the BRCA gene of patients with ovarian cancer were collected and mutation site analysis was performed to compare phenotypic differences in pathogenic mutations between HBOC syndrome and SOC patients.Results:(1) Of the 284 ovarian cancer patients, seventy-seven had BRCA pathogenic mutations with a mutation rate of 27.1% (77/284), with BRCA1 mutation rate of 19.7% (56/284), BRCA2 gene 6.7% (19/284) and BRCA1/2 common mutation rate of 0.7% (2/284). Of the 284 patients with ovarian cancer, the pathogenic mutation rate in the BRCA gene in HBOC syndrome patients was 43.8% (32/73), which were significantly higher than that in SOC patients [21.3% (45/211); χ2=13.905, P<0.01]. Among BRCA1 gene mutation, the mutation rate in HBOC syndrome was higher than that of SOC [87.5% (28/32) vs 62.2% (28/45)], the BRCA2 gene mutation rate in patients with HBOC syndrome was lower than that in SOC patients [6.2% (2/32) vs 37.8% (17/45)], and there were statistically significant differences (all P<0.05). Two of the 77 patients with pathogenic mutations in the BRCA gene were multisite mutations, including one simultaneous two site mutation, one simultaneous three site mutation. There were 80 mutation sites with frameshift deletion mutations (55.0%, 44/80) and nonsense mutations (31.2%, 25/80). (2) Of the 73 patients with HBOC syndrome, 32 cases had pathogenic mutations in BRCA gene, including 28 cases in BRCA1, mainly in exon 11 and 24 (9 and 7 cases, respectively), and only two cases in BRCA2, both in exon 11; another two had multiple locus mutations. Of the 211 patients with SOC, 45 cases had pathogenic mutants in BRCA gene, including 28 cases in BRCA1, mainly in exon 11 and 24 (15 and 2 cases, respectively), and 17 cases in BRCA2, mainly in exon 11 (11 cases). (3) Thirty-four pathogenic mutation sites in BRCA gene were found newly, twenty of them were located in the BRCA1 gene, including a locus located on the intron 6, 301+1G>A, and the remaining 19 sites were located on the exons, including 283_286delCTTG, 68_69delAG, 132C>T, 514_547+3del37, 742delA, 1126_1129delAATA, 1196delA, 1352_1364del, 1465G>T, 2171delC, 2341G>T, 3359_3363delTTAAT, 4085_4086ins11, 4161_4162delTC, 4165_4166delAG, 4258G>T, 4338_4339del8insAGAA, 4468G>T, and 4783delA; fourteen sites were located in the BRCA2 gene, including a locus located on the intron 7, 631+1G>A, and the remaining 13 sites were located on the exons, including 2648delT, 2914A>T, 2950_2951insG, 4357+1G>A, 5054C>T, 5257A>T, 5291_5292insTC, 5913delT, 3593delA, 6091_6092insA, 6135_6136delTT, 7452delT, 9097_9098insA. A tal of 28 repeat mutations were located in the BRCA1 gene; among them, the site 5470_5477del8 was repeated 6 times, while 3 times in 981_982delAT. Conclusions:Patients with HBOC syndrome have a significantly higher rate of pathogenic mutation in the BRCA gene than that in patients with SOC. BRCA gene pathogenic mutation sites in HBOC syndrome patients occur commonly in exon 11 and 24 of BRCA 1 gene, while SOC patients occur mainly in exon 11 and 24 of BRCA1 gene and exon 11 of BRCA2 gene. The two loci of BRCA1∶5470_5477del8, BRCA1∶981_982delAT may be ancestor mutations in Chinese ovarian cancer patients, and 34 newly discovered pathogenic mutations in the BRCA gene, enriching the BRCA gene mutation spectrum in the Chinese population.

The aim of this study was to investigate the effect of transmembrane 9 superfamily protein member 2 (TM9SF2) in proliferation and migration of triple negative breast cancer cell line MDA-MB-231.The expression of TM9SF2 in triple negative breast cancer cell line MDA-MB-231 and nontumorigenic mammary epithelial cell line MCF-10A were measured by Western blot. MDA-MB-231 cells were treated with siRNA-TM9SF2 to knock-down the expression of TM9SF2. The effect of silencing TM9SF2 was measured with Western blot.The proliferation of cells was tested by MTS,and the migration was measured with Transwell and wound-healing assay.Proteins related to proliferation (PI3K,AKT,SRC and ERK) and migration (Snail,Slug and N-cadherin) were measured with Western blot.Protein expressions of TM9SF2 was better improved in triple negative breast cancer MDA-MB-231 cell line than MCF-10A.Compared with the control group, the siRNA-TM9SF2 infected group had lower expressions of PI3K, Snail, Slug and N-cadherin, and at the same time phosphorylation of AKT was decreased. The results suggest TM9SF2 can promote the proliferation and metastasis of triple negative breast cancer MDA-MB-231 cell line.