In this study, the fatty acid desaturase gene FAD2 was cloned from Coix lacryma-jobi L. and its molecular structure and function were studied. The results showed that the full-length cDNA sequence of FAD2 gene was 936 bp encoding 311 amino acid residues. Bioinformatics prediction results showed that the protein encoded by the FAD2 gene was an alkaline hydrophilic unstable protein with a molecular weight of 34.87 kDa. It contained three transmembrane helix domain, and did not contain the signal peptide splicing site, and was most likely to be located in plasmid membrane. Compared with other similar genes in plants, it has only a histidine conserved site, His Box Ⅲ histidine site (HXXHH), suggesting its activity may be reduced. Phylogenetic tree analysis showed that FAD2 was closely related to monocotyledonous plants, especially Maize and Oryza sativa japonica Group, but farther from dicotyledonous plants. Therefore, it was inferred that FAD2 might have similar functions with similar genes in Maize and Oryza sativa japonica Group. In addition, the expression of FAD2 gene could be detected in Coix lacryma-jobi L. with high oil content, but not in low oil content of Coix lacryma-jobi L. In order to clarify the function of FAD2, the gene was heterologously expressed in sporomyces cerevisiae. The results showed that the protein encoded by FAD2 gene did not catalyze the formation of C18∶1 unsaturated fatty acid into C18∶2 unsaturated fatty acid. Therefore, it was speculated that the deletion of histidinine conserved site of FAD2 gene might lead to the decrease of protein activity or even inactivation. This study provides reference value for further understanding the molecular structure characteristics of fatty acid desaturase. At the same time, it laid a foundation for elucidating the biosynthetic pathway of Coix lacryma-jobi L.

Objective To review the existing literature and quantitatively evaluate the association of circulating vaspin levels and the risk of gestational diabetes mellitus ( GDM) ． Methods We systematically searched the PubMed，EMBASE，Web of Science，China National Knowledge Infrastructure，and WanfangData databases up to June 2019． Pooled standardized mean differences ( SMDs) with 95% confidence intervals ( CIs) were calculated using random－ or fixed－effects models based on the heterogeneity of studies． Subgroup analyses，Meta－regression，sensitivity and publication bias were assessed to analyze the heterogeneity and the robustness of the results． All statistical analyses were performed using STATA 12．0． Ｒesults Nine articles ( 11 comparisons) published from 2013 to 2019 were included in our final Meta－analysis，covering a total of 738 patients with GDM and 661 normal pregnant women． There was significant difference in the overall maternal circulating vaspin levels between GDM patients and healthy pregnant women ( SMD= 0．613，95% CI: 0．044－1．182，P= 0．035) ． Subgroup analyses stratified by trimester in which vaspin was measured and whether BMI was matched suggested the similar trend to the overall result． Subgroup analysis according to ethnicity found that circulating vaspin level might not be related to GDM in " European" subgroup; sensitivity analysis by excluding moderate－quality studies and BMI－unmatched studies found that circulating vaspin levels were still related to GDM risk． Conclusions Our Meta－analysis indicated that maternal circulating vaspin levels might be positively correlated with the risk of GDM in Asians．

Prohibitin (PHB), an evolutionarily conserved mitochondrial inner membrane protein, is highly expressed in cells that require strong mitochondrial function. Recently, we demonstrated that the deletion of Phb in spermatocytes results in impaired mitochondrial function. In addition, PHB expression in the mitochondrial sheath of human sperm has a significantly negative correlation with mitochondrial reactive oxygen species levels, but a positive one with mitochondrial membrane potential and sperm motility. These results suggest that mitochondrial PHB expression plays a role in sperm motility. However, the mechanism of PHB-mediated regulation of sperm motility remains unknown. Here, we demonstrate for the first time that PHB interacts with protein kinase B (AKT) and exists in a complex with phospho-PHB (pT258) and phospho-AKT in the mitochondrial sheath of murine sperm, as determined using colocalization and coimmunoprecipitation assays. After blocking AKT activity using wortmannin (a phosphatidylinositol 3-kinase [PI3K] inhibitor), murine sperm have significantly ( P < 0.05) decreased levels of phospho-PHB (pT258) and the total and progressive motility. Furthermore, significantly ( P < 0.05) lower levels of phospho-PI3K P85 subunit α+γ (pY199 and pY467) and phospho-AKT (pS473; pT308) are found in sperm from infertile asthenospermic and oligoasthenospermic men compared with normospermic subjects, which suggest a reduced activity of the PI3K/AKT pathway in these infertile subjects. Importantly, these sperm from infertile subjects also have a significantly ( P < 0.05) lower level of phospho-PHB (pT258). Collectively, our findings suggest that the interaction of PHB with AKT in the mitochondrial sheath is critical for sperm motility, where PHB phosphorylation (pT258) level and PI3K/AKT activity are key regulatory factors.

Objective:To summarize the application of phage therapy in patients with urinary tract complicated pandrug-resistant Klebsiella pneumoniae infection, and analyze its feasibility and effectiveness.Methods:To retrospectively analyze the clinical data of a patient with complicated urinary tract complex pan-resistant Klebsiella pneumoniae treated by phage from August to September, 2019 in Shanghai Public Health Clinical Center. The female patient, 65 years old, was admitted to the hospital on August 6, 2020. The patient repeated with frequent micturition and urgent micturition half a year before admission. These symptoms were not accompanied by back pain, fever, chills, dysuria, gross hematuria. Urinary culture results in outpatient hospital was pan-resistant Klebsiella pneumoniae. After the patient discontinued application of cefoperazone sulbactam, levofloxacin and other drugs, symptoms such as frequent urination could be relieved after treatment, but appeared repeatedly. In August 2019, the center innovatively applied phage therapy to treat this patient with urinary tract pandrug-resistant bacteria infection.Results:For the first time, we applied 117, 135, 178, GD168 phage mixed solution once a day, for 5 days of continuous bladder infusion. At the same time, meropenem and amikacin was intravenous administration to strengthened the anti-infection treatment. Urine culture was negative for two consecutive times after treatment. However, half a month after the end of the bladder infusion, the patient experienced discomfort such as frequent urination. Urine culture: pan-resistant Klebsiella pneumoniae. The second time, we applied a mixture of three phage strains 130, 131, 909, once a day, for 5 days of continuous bladder infusion. And in the afternoon of the third day of treatment, the renal pelvis was retrogradely intubated and perfused with the above three strains of phage mixture. During the second treatment follow-up until March 30, 2020, the patient's urine culture was reviewed once a month. As a result, no pan-resistant Klebsiella pneumoniae was found, and the patient no longer experienced frequent urination and other symptoms of urination. The treatment process was successful and without severe complications and side effects.Conclusions:Phage urinary tract perfusion is an effective method for the treatment of pan-resistant Klebsiella pneumoniae urinary tract infections. The curative effect is accurate and reliable. The patient did not show obvious complications and adverse reactions during treatment. It can be used as an alternative treatment plan for complex pan-resistant Klebsiella pneumoniae infection.

Development of rapid analytical methods and establishment of toxic component limitation standards are of great importance in quality control of traditional Chinese medicine. Herein, an on-line extraction electrospray ionization mass spectrometry (oEESI-MS) coupled with a novel whole process integral quantification strategy was developed and applied to direct determination of nine key aconitine-type alkaloids in 20 proprietary Chinese medicines (APCMs). Multi-type dosage forms (, tablets, capsules, pills, granules, and liquid preparation) of APCM could be determined directly with excellent versatility. The strategy has the characteristics of high throughput, good tolerance of matrix interference, small amount of sample (∼0.5 mg) and reagent (∼240 μL) consumption, and short analysis time for single sample (<15 min). The results were proved to be credible by high performance liquid chromatography-mass spectrometry (LC-MS) and electrospray ionization mass spectrometry, respectively. Moreover, the limitation standard for the toxic aconitines in 20 APCMs was established based on the holistic weight toxicity (HWT) evaluation and the severally, and turned out that HWT-based toxicity evaluation results were closer to the real clinical applications. Hence, a more accurate and reliable APCM toxicity limitation was established and expected to play an important guiding role in clinics. The current study extended the power of ambient MS as a method for the direct quantification of molecules in complex samples, which is commonly required in pharmaceutical analysis, food safety control, public security, and many other disciplines.

OBJECTIVE: To investigate the effect of other gene mutations outside the fusion gene on the first complete remission (CR) induced by one course of induction chemotherapy in patients with core binding factor-associated acute myeloid leukemia (CBF-AML). METHODS: DNA was extracted from bone marrow or peripheral blood samples of newly diagnosed CBF-AML patients admitted to the Hematology Department of the Second Hospital of Shanxi Medical University from January 2015 to January 2019. Next-generation sequencing was used for detection of 34 kinds of hematologic malignancy-related gene mutations in patients with CBF-AML, the effect of related gene mutations on the first complete remission (CR) rate in one course of induction chemotherapy was analyzed by combineation with clinical characteristics. RESULTS: 34 kinds of genes in bone marrow or peripheral blood of 43 patients were detected by high throughput sequencing and the gene mutations were detected in 16 out of 34 genes. The mutation rate of KIT gene was the highest (48.8%), followed by NRAS (16.3%), ASXL1 (16.3%), TET2 (11.6%), CSF3R (9.3%), FLT3 (9.3%), KRAS (7.0%). The detection rates of mutations in different functional genes were as follows: genes related with signal transduction pathway (KIT, FLT3, CSF3R, KRAS, NRAS, JAK2, CALR, SH2B3, CBL) had the highest mutation frequency (72.1% (31/43); epigenetic modification gene mutation frequency was 30.2% (13/43), including ASXL1, TET2, BCOR); transcriptional regulation gene mutation frequency was 7.0% (3/43), including ETV6, RUNX1, GATA2). Splicing factor related gene mutation frequency was 2.3% (1/43), including ZRSR2). The CR rate was 74.4% after one course of induction chemotherapy. At first diagnosis, patients with low expression of WT1 (the median value of WT1 was 788.9) were more likely to get CR (P=0.032) and the RFS of patients who got CR after one course of induction chemotherapy was significantly longer than that of patients without CR [7.6 (2.2-44.1) versus 5.8 (1-19.4), (P=0.048)]. The rate of CR in the signal transduction pathway gene mutation group was significantly lower than that in non-mutation group (64.5% vs 100%) (P=0.045), while the level of serum hydroxybutyrate dehydrogenase (HBDH) was significantly higher than that in non-mutation group [(418 (154-2702) vs 246 (110-1068)] (P=0.032). There was no difference in CD56 expression between the two groups (P=0.053), which was limited to the difference between (≥20%) expression and non-expression. (P=0.048). CONCLUSION CBF-AML patients with signal transduction pathway gene mutation are often accompanied by high HBDH level and CD56 expression, moreover, the remission rate induced by one course of treatment is low.
High-Throughput Nucleotide Sequencing ; Humans ; Leukemia, Myeloid, Acute ; Mutation ; Prognosis ; Signal Transduction