A temperature-sensitive mutant strain was isolated after transposon mutagenesis with Tn5 and named MT54.PCR was carried out with primers designed according to the sequence of transposon,the PCR products showed that the MT54 carried transposon in the genome. The mutant grew well at 30℃in minimal medium(MM)containing p-nitrophenol(PNP)as sole carbon source,while it cannot grow at 37℃in the same medium,NO_2.detection results also proved that.Comparing the degradation rate of PNP and hydroquinone of MT54 and DLL-E4 at different temperature,it was speculated that the mutant site locate in the PNP degradation related genes.

Objective To investigate the therapeutic effects of electroacupuncture (EA) on post stroke depres- sion (PSD) after lacunar infarction (LI).Methods Seventy-five PSD patients with lacular infarction were recruited and randomly divided into a control group and an EA group.All patients were treated with Fluoxetine,in addition to EA treat- ment in the EA group.Then all patients were evaluated using the Hamilton Depression Scale (HAMD),the Chinese stroke scale (CSS) and the Barthel Index (BI).The evaluations were carried out at 0,14 and 28 days.Results Depression symptoms (DSs) in the EA group were improved by day 14 of the treatment,and their HAMD scores had decreased.DSs in both groups had improved significantly by day 28,and the HAMD scores in the EA group were then significantly lower than those in the control group.Improvements in neurological impairment and in the activities of daily living were observed earlier in the EA group.Conclusion The combination of EA and Fluoxetine is helpful for PSD with complex patho- genesis and clinical symptoms.Therapeutic effects are enhanced and side effects are reduced.

Objective To investigate Glypican-3 gene expression and mutation in hepatocellular carcinoma (HCC).Methods Glypican-3 gene expression and mutation in tumor,para-c.ancer and normal tissue of 48 HCCs were detected by RT-PCR and single-strand conformation polymorphism(SSCP),respectively.Results There was no Glypican-3 mRNA expression in para-cancer and normal tissue.Expression rate of Glypican-3 mRNA was 77.1% in tumor tissue,which was correlated with clinical staging and cell differentiation(P

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

AIM:To explore the role of sphingosine 1-phosphate (S1P) in the dysfunction of vascular endo-thelial cells exposed to high glucose.METHODS: In human aortic endothelial cells cultured under high-glucose ( 22 mmol/L glucose) medium, nitric oxide ( NO) level, polymorphonuclear neutrophil-endothelial cell adhesion rate, protein level of intercellular adhesion molecule-1 ( ICAM-1) , migration of endothelial cells and Akt/endothelial nitric oxide syn-thase ( eNOS) pathway activation were observed after S1P, sphingosine kinase-1 inhibitor and/or Akt inhibitor treatments. RESULTS:S1P decreased NO level, increased polymorphonuclear neutrophil adhesive rate, enhanced ICAM-1 protein level, and inhibited migration of endothelial cells and activation of Akt/eNOS pathway in endothelial cells cultured under high-glucose condition.Sphingosine kinase-1 inhibitor, which reduced S1P content, significantly improved the above endo-thelial cell function indexes and restored the activation of Akt/eNOS pathway.CONCLUSION: S1P promoted high glu-cose-induced dysfunction of endothelial cells probably by inhibiting the activation of Akt/eNOS signal pathway.Targeting S1P is expected to become one of potential treatment strategies to reduce endothelial cell dysfunction.

Lysine-specific demethylase 1 (LSD1) plays vital roles in cell stemness, differentiation, cell motility, metabolic control and epithelial-mesenchymal transition, which is closely associated with tumorigenesis processes including cell proliferation, invasive, metastasis and poor prognosis. Besides, LSD1 also contributes to the occurrence of other diseases such as neurodegenerative diseases and viral infections. Since 2013, the irreversible inhibitors including tranylcypromine, ORY-1001, ORY-2001, GSK-2879552, IMG-7289, INCB059872, TAK-418, LH-1802 and reversible inhibitors including CC-90011 and SP-2577 have been approved for clinical assessment. This review comprehensively summarizes the clinical research of LSD1 drug candidates and briefly discusses the prospects, opportunities and challenges of LSD1-targeted drug discovery, aiming to provide a landscape for the related drug development.

Abstract: Objective　By analyzing the frequency distribution of antihypertensive drug-related genotypes in hypertensionpatients treated in our hospital, so as to provide a clinical basis for individualized treatment of hypertension patients. Methods　A total of 72 hypertensive patients treated in Hainan Hospital of PLA General Hospital from June 2021 to April 2022 were collected. PCR-melting curve method was used to detect CYP2D6*10 (c.100 C>T), CYP2C9*3 (c.1075 A>C), ADRB1 (c.1165 G>C), AGTR1 (c.1166 A>C), ACE (I/D), NPPA (T2238C) and CYP3A5*3 (A6986G), and the relationship between different genotypes and biochemical indexes was analyzed. Results　According to the statistics of the gene and genotype frequency of each point in 72 patients, the gene frequencies of 7 sites all conformed to Hardy Weinberg equilibrium. There were gender differences in ADRB1 genotypes (χ2 = 5.878, P<0.05). There were statistical differences in triglycerides [AA: 1.4 (1.0, 2.0)mmol/L; AC: 2.2 (1.5, 2.5)mmol/L; P=0.038], total cholesterol [AA: 4.0 (3.1, 4.9) mmol/L; AC: 4.8 (4.0, 5.3) mmol/L; P=0.040] and low-density lipoprotein cholesterol [(AA: 2.4 (1.8, 3.3) mmol/L; AC: 3.2 (2.5, 3.5) mmol/L; P=0.035] among patients with different genotypes of AGTR1 locus. The patients with different genotypes of CYP2C9 locus had significant differences in their alanine transferase (ALT) [AA:16.9 (11.4,30.2) mmol/L; AC:10.4 (9.4, 18.2) mmol/L; P=0.040]. Aftergene-directed individualized therapy, different genotypes of CYP3A5 andAGTR1 affected the heart rate [CYP3A5: AA: (79.3±7.0) beats/min; AG： (69.8±6.8) beats/min; GG： (68.8±7.3) beats/min; P=0.010], systolic blood pressure [AGTR1: AA: (131.3±16.7) mmHg; AC: (140.6±11.8) mmHg; P=0.014] and diastolic blood pressure [CYP3A5: AA: (90.0±8.3) mmHg; AG: (78.7±10.8) mmHg; GG: (74.9±10.7) mmHg; P=0.025; AGTR1: AA: (75.3±10.2) mmHg; AC: (86.3±10.6) mmHg; P=0.001] of patients. Conclusions　The related gene loci of antihypertensive drugs are an important basis for guiding the diversification and individualization of clinical medication. Clinicians need to consider the impact of related genes on drug efficacy and adverse reactions when prescribing.

Objective To evaluate the CT findings of pancreatic carcinoid tumors.Methods The CT imaging data of five patients with pancreatic carcinoid tumors confirmed by pathology were retrospectively analyzed.Results The tumors ranged in maximum diameter from 2.0 to 11.0 cm with a mean of 6.4 cm. On unenhanced CT,the tumors were slightly hypodense relative to the pancreatic parenchyma,homogenous in 2 cases,and heterogenous in 3 cases.One tumor showed calcification.After contrast material injection, the solid component of the tumor showed marked heterogenous enhancement on the arterial phase scanning in 3 cases,and mild heterogenous enhancement in 2 cases.The degree of tumor enhancement was less intense than the surrounding pancreatic parenchyma due to necrosis of various degree,which led to the cystic appearance of the tumor in 1 ease.On the portal phase scanning,all tumors showed marked enhancement similar to that of the pancreatic parenchyma.On the delayed phase scanning,the degree of enhancement was more intense than the surrounding pancreatic parenchyma in 1 case.Liver metastases with retroperitoneal lymphadenopathy and peripancreatic vessels invasion were seen in 1 case.No dilatation of the biliary tract or pancreatic duct was present.Conclusion The CT features of pancreatic carcinoid tumors included infrequent dilatation of the biliary tract or pancreatic duct and unusual vascular involvement,calcification within the mass,marked enhancement similar to that of the surrounding pancreatic parenchyma during the portal phase scanning and more intense during the delayed phase scanning.

AIM To explore the action mechanism of puerarin's protective effects against oxidative stress of HUVEC-12 cells induced by high glucose.METHODS HUVEC-12 cells cultured with 100 mmol/L glucose medium and 10,25,50 μmoL/L puerarin for 36 h had the cell proliferation,the levels of lactate dehydrogenase (LDH),intracellular reactive oxygen species (ROS),the activities of caspase-3,superoxide dismutase (SOD)and catalase (CAT),and the contents of malondialdehyde (MDA) and glutathione (GSH) measured.The mRNA expressions of SIRT1 and PGC-1α were detected by real-time flu orescence quantitative PCR,and the protein contents of SIRT1 and PGC-1 α were determined by enzyme-linked immunosorbent assay.RESULTS The puerarin treatment to HUVEC-12 cells resulted in markedly lowered LDH level,caspase-3 activity,intracellular levels of MDA and ROS,and notable improvement of the cell viability,the activities of SOD and CAT,GSH content,the mRNA expressions and the protein contents of SIRT1 and PGC-1α as well.CONCLUSION The protective effect of puerarin on high glucose-induced oxidative damage of HUVEC-12 cells may be attributed to the SIRT1/PGC-1α pathway activation.

OBJECTIVETo investigate the influence of cigarette smoking on human sperm DNA integrity.

METHODSTotally, 784 cases of male infertility were selected from our case database and grouped according to whether they were smokers or nonsmokers, how much they smoked (< or = 10, 11-19 and > or = 20 cigarettes/d) and how long they smoked (< or = 5, 6-9 and > or = 10 yr). Sperm DNA integrity was measured using sperm chromatin structure assay (SCSA) and flow cytometry. DNA fragmentation and immature spermatozoa were expressed by the DNA fragmentation index (DFI) and high DNA stainability (HDS) respectively. Conventional sperm parameters and sperm DNA integrity were compared among different groups.

RESULTSThe total semen volume and percentage of grade a + b sperm were lower and the sperm morphological abnormality was higher in the > or = 20 cigarettes/d and > or = 10 yr groups than in the others (P < 0.05). DFI and HDS were significantly higher in the smokers than in the nonsmokers (P < 0.05). HDS was negatively correlated with the percentage of grade a + b sperm (r = -0.18, P < 0.05) and both DFI and HDS were positively correlated with the rate of sperm malformation (r = 0.31 and r = 0.39, P < 0.05).

CONCLUSIONSmoking more than 20 cigarettes a day or longer than 10 years has deleterious effects on the semen volume, percentage of grade a + b sperm and sperm morphology of the smokers. Cigarette smoking decreases sperm DNA integrity and nuclear maturation.

Adult ; DNA Damage ; drug effects ; DNA Fragmentation ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Smoking ; adverse effects ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; Young Adult