Objective To evaluate the relationship between spectinomycin resistance in Neisseria gonorrhoeae and mutations in the rpsE gene.Methods Genomic DNA was extracted from 4 clinical isolates of Neisseria gonorrhoeae with different levels of spectinomycin resistance.Then,PCR was performed to amplify the entire rpsE gene and the spectinomycin resistance-determining region (SRDR) in the 16S rRNA gene followed by direct sequencing.Two spectinomycin-sensitive Neisseria gonorrhoeae strains were transformed with the genomic DNA containing the mutant rpsE gene.Subsequently,the susceptibility of the transformants to spectinomycin was determined,and PCR was performed to amplify the rpsE and 16S rRNA genes in the transformants followed by sequencing.Results All the 4 spectinomycin-resistant Neisseria gonorrhoeae strains harbored an A70C transversion in the rpsE gene,but no abnormality in the SRDR of the 16S rRNA gene.No mutations were detected in the spectinomycin-sensitive Neisseria gonorrhoeae strains.The A70C transversion in the rpsE gene was also detected in the two Neisseria gonorrhoeae transformants with spectinomycin resistance.Conclusion The A70C point mutation within the rpsE gene is associated with spectinomycin resistance in Neisseria gonorrhoeae.

Objective To discuss the specific role of association rules in the compatibility of herbs and the relationship between herbs and symptoms by analyzing ancient analogous prescriptions Zhizichi Decoction with SQL Server software.Methods The prescriptions which contain Gardenia and Semen Sojae Preparatum were searched in the database of “Ancient Prescriptions for Epidemic Disease”, and they were imported to software with the standardized data of their symptoms and ingredients, then the proper association rules were found. The laws between parameter setting and rule generation were searched through parameter adjustment.Results Many laws of the drug compatibility and relationship between herbs and symptoms were obtained effectively in data mining by adjusting the parameters:The support setting was good for finding common used drug compatibility, and the confidence was good for finding specific drug compatibility and relationship between herbs and symptoms;the item set which had high frequency, sometimes made the rule’s importance low.Conclusion The laws of compatibility of herbs and the relationship between herbs and symptoms for data mining of analogous prescriptions are discovered by analyzing the support, confidence and importance of the association-rules and the clinical magnificence between symptoms and herbs.

This paper is to report the exploration of the activation of Rho/ROCK signal pathway in 5-HT-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and the inhibitory effect of m-Nis on this pathway. PASMCs were cultured with the explant technique. MTT assay was used to explore the proliferation of PASMCs after 5-HT treated for different time and the intervening effect of m-Nis. RT-PCR and Western blot were used respectively to explore the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 in 5-HT-treated PASMCs and intervening effect of m-Nis. The results of MTT assay suggested that 5-HT (1 µmol · L(-1)) treatment for 12-72 h significantly induced the proliferation of rat PASMCs (P<0.05 or P < 0.01), which were inhibited by m-Nis (1 x 10(-5), 1 x 10(-6), l x 10(-7), 1 x10(-8) mol · L(-1)) in dose-dependent manners (P < 0.05 or P < 0.01). Similarly, the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 were also inhibited by m-Nis in different degrees (P < 0.05 or P < 0.01). Thus, the results of this study suggested that Rho/ROCK pathway played an important role in 5-HT-induced proliferation of rat PASMCs, m-Nis inhibited 5-HT-induced proliferation obviously, which may be related to the blockage of Rho/ROCK signal pathway.
Animals ; Cell Proliferation ; drug effects ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Nisoldipine ; pharmacology ; Protein Phosphatase 1 ; metabolism ; Pulmonary Artery ; cytology ; Rats ; Serotonin ; pharmacology ; Signal Transduction ; rho-Associated Kinases ; metabolism ; rhoA GTP-Binding Protein ; metabolism

Aim To investigate the effect of intrathecal injection(it) of minocycline(MC),a selective microglia inhibitor,on the maintenance of pain in a murine model of cancer pain.Methods Forty-two male C3H/He mice were randomly divided into 3 groups(n=14 each):sham+PBS group was operated and received 10 ?l PBS injection into calcaneus medullary space;sarcoma+PBS group and sarcoma+MC group were operated and received 2?105 sarcoma cells/10 ?l PBS implantation into calcaneus medullary space.On the PID11(post-implantation day 11),10 mice were taken from each group randomly,sarcoma+PBS group and sham+PBS group were received 0.9% NS(5 ?l) it sarcoma+MC group was received MC(1 nmol,5 ?l) it Mechanical pain threshold and cold hyperalgesia assay were measured before and after it at 0.5,1,2,4,8,24 h.The last 4 mice of each group were received a normally non-noxious palpation of the ipsilateral heel 90 min on the PID12 after it,then the animals were killed and L4-6 segment of spinal cord was removed for analysing the c-fos expression(by immunohistochemistry).Results Bone cancer pain decreased the mechanical and cold pain threshold and activated the c-fos expression in the spinal cord;MC it transient attenuated bone cancer pain-induced hyperalgesia and allodynia and suppressed the expression of c-fos protein.Conclusion The activation of microglia in the spinal cord may be involved in the maintenance of bone cancer pian.

OBJECTIVETo explore the anesthetic effect of preemptive analgesia of frequency acupoint electrical stimulation on painless-induced abortion as well as its effect on anesthetics dosage.

METHODSNinety cases of early pregnancy who selected painless-induced abortion were randomly divided into two groups, 45 cases in each group. Frequency acupoint electrical stimulation at Ciliao (BL 32) and Shenshu (BL 23), disperse-densewave, 2 Hz/100 Hz in frequency for 15 to 20 min, was applied in the group A, which was followed by intravenous anesthesia of propofol. The intravenous anesthesia of propofol was applied in the group B. The blood pressure (BP), heart rate (HR) and SpO2 before, during and after surgery, anesthetic effect and dosage, waking time and adverse events were observed in the two groups.

RESULTSThe BP and HR during and after the surgery in the group A were not statistically different from those before the surgery (all P > 0.05). The BP was reduced and HR was slowed down during the surgery in the group B, which was significantly different from those before the surgery as well as those in the group A (all P < 0.05). The dosage of propofol was (114. 3-+6. 1) mg in the group A. obviously less than (193.2 +/- 8.9) mg in the group B (P < 0.05). The waking time was (5.6 +/- 1.2) min in the group A, obviously less than (10.1 +/- 3.9) min in the group B (P < 0.05). As for anesthetic effect, the incidence of Grade I in the group A was more than the group B (P < 0.05). The adverse events, including nausea, vomiting and contractions pain in the group A were evidently less than those in the group B (all P < 0.05).

CONCLUSIONThe preemptive analgesia of frequency acupoint electrical stimulation could significantly improve anesthetic effect of painless-induced abortion, reduce dosage of anesthetics, shorten waking time of surgery and guarantee the safety of surgery.

Abortion, Induced ; Acupuncture Analgesia ; Acupuncture Points ; Adult ; Electric Stimulation ; Female ; Humans ; Pain Management ; Pregnancy ; Young Adult

BackgroundDiabetic retinopathy (DR) is the result of the cytokine network disorders,the imbalance of angiogenic factor and vascular inhibitory factor is the start factor.ObjectiveTo analyze the levels of CXC chemotatic factors of type 2 diabetes mellitus patients,evaluate the clinical application value of them in different clinical types of DR using receiver operating characteristic (ROC)analysis and to approach the new way of individualized treatment.Methods This was a prospective research.The gold standard was ophthalmolscope and fundus fluorescein angiography.The levels of CXC chemotatic factors and multiplicaiton factors were measured in 96 cases with type 2 diabetes mellitus (66 cases with retinopathy and 30 cases without retinopathy as control).The assessment tasks were performed for these index and courses of DR with ROC curve.Results The expression of age,course of disease has significant difference in different courses of DR ( F =8.507,P =0.001 ; F =28.143,P =0.000).Compared with the control group,the expression of growth-related oncogene-α ( GROα ) ( t =- 2.172,P =0.035,AUC =0.625 ),whole blood viscosity 200 ( t =- 3.724,P =0.001,AUC =0.904 ) and neutrophilic leukocyte (t=-2.562,P =0.013,AUC =0.577 ) has significant difference in the group of mild NPDR.Compared with the control group,the expression of interferon-γ-inducible protein 10 ( IP-10 ) ( t =-3.591,P =0.001,AUC =0.592 ),platelet derivation growth factor-BB ( PDGF-BB ) ( t =- 3.233,P =0.003,AUC =0.735 ),vascular endothelial growth factor(VEGF) ( t =- 3.617,P =0.001,AUC =0.776 ),C peptide ( t =- 3.366,P =0.002,AUC =0.962 ),leukocyte ( t=-3.201,P =0.003,AUC =0.852) and neutrophilic leukocyte(t =-4.201,P=0.000,AUC =0.852) has significant difference in the group of moderate and severe NPDR.ConclusionsCXC chemotatic factors may act as reactivator in the pathogenesis of DR,GROα and IP-10 may be useful for clinical monitoring of the severity of DR,and evaluating the imbalance state of chemotatic factors maybe a new approach to clinical monitoring and prognosis of DR.

S-adenosylmethionine-dependent uroporphyrinogen III methyltransferase (SUMT) is a novel red fluorescence indicator. However, the production of SUMT in Escherichia coli is restricted by its relatively low solubility, and little is known about the red fluorescent materials that are associate with SUMT. Two individual SUMT mutations, L166A and L88R/L89G double mutant were produced by site-directed mutagenesis. Both mutants were overexpressed in E. coli and purified by Ni-NTA chromatography. The reddish mixtures isolated from the purified L88R/L89G double mutant were analyzed by UV-visible spectra scanning and mass analysis(MS). The L88R/L89G double mutant has enzymatic activity in vivo, whereas L166A mutant loses the activity. Trimethylpyrrocorphin is identified as the main constituent in the isolated pigments. The purified L88R/L89G mutant increases protein solubility, which is applied potentially as the fluorescent indicator denoting the solubility of protein fusion partner.
Amino Acid Substitution ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Fluorescence ; Mass Spectrometry ; Methyltransferases ; chemistry ; genetics ; metabolism ; Molecular Weight ; Mutagenesis, Site-Directed ; methods ; Mutation ; Pigments, Biological ; chemistry ; metabolism ; Plant Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; chemistry ; metabolism ; Solubility ; Spectrophotometry, Ultraviolet ; Zea mays ; enzymology ; genetics

Recently, novel Psidium meroterpenoids were reported in the guava leaves. According to careful analysis of the spectral data of literatures, the spectroscopic characteristics and biosynthetic pathway of Psidium meroterpenoids were summarized in this paper. The results showed that Psidium meroterpenoids had distinct spectroscopic features and reasonable biosynthetic routines, however the number order of carbon atoms was not consistent in the reported literatures. It was concluded that Psidium meroterpenoids were the characteristic chemical constituents of Psidium guajava Linn.
Magnetic Resonance Spectroscopy ; Plant Leaves ; chemistry ; Psidium ; chemistry ; Spectrum Analysis ; Terpenes ; chemistry

Objective To investigate and compare the different effects of soluble adult wornl antigen(SWA)and soluble egg antigen(SEA)of Schistosoma japonicum on the apoptosis and cell-cycle of routine CD4~+T cells.Methods Purified CD4~+T ceUs from normal C57BL/6 mice were cultured with CFSE labeled antigen presenting clls in the presence of different stimuli for 36 h.Flow cytometry(FCM)was used to detect the apoptosis of CD4~+T cells by fluorescence conjugated caspase-3 antibodie staining.The flow cytometry was used to analyze the cell-cycle of CD4~+T cells cultured as described above for 96 h by propidium iodide staining.Results Compared with the apoptosis percentage of CD4~+T cells[(1.24±0.29)%]in the SEA stimulated group,that in the SWA stimulated group[(1.52±0.38)%]did not show statistically significant difference(P>0.05).Compared with the cell percentages in G1 phase[(78.91±2.98)%],S phase[(7.39±0.85)%]and G2/M phase[(10.69±1.05)%] in the SWA stimulated group,that of the G1 phase[(59.42±1.32)%]was significantly lower,but those in the S phase[(21.07±O.88)%] and G2/M phase[(18.88±1.21)%]were significantly increased in the SEA stimulated group(P<0.01).Conclusions There is no statistically significant difference between the apoptosis levels of CD4~+T ceHs stimulated by SWA and SEA.However,SEA significantly promotes the progression of the cell-cycle of CD4~+T cells compared with SWA.

Most chemical medicines have polymorphs. The difference of medicine polymorphs in physicochemical properties directly affects the stability, efficacy, and safety of solid medicine products. Polymorphs is incomparably important to pharmaceutical chemistry, manufacturing, and control. Meantime polymorphs is a key factor for the quality of high-end drug and formulations. Polymorph prediction technology can effectively guide screening of trial experiments, and reduce the risk of missing stable crystal form in the traditional experiment. Polymorph prediction technology was firstly based on theoretical calculations such as quantum mechanics and computational chemistry, and then was developed by the key technology of machine learning using the artificial intelligence. Nowadays, the popular trend is to combine the advantages of theoretical calculation and machine learning to jointly predict crystal structure. Recently, predicting medicine polymorphs has still been a challenging problem. It is expected to learn from and integrate existing technologies to predict medicine polymorphs more accurately and efficiently.