Structural Analysis and Characterization of 4-F-α-PVP Analog 4-F-3-Methyl-α- PVP Hydrochloride / 法医学杂志

OBJECTIVES@#To identify 1-(4-fluorophenyl)-2-(1-pyrrolidinyl) pentan-1-one (4-F-α-PVP) analog 1-(4-fluoro-3-methyl phenyl)-2-(1-pyrrolidinyl) pentan-1-one (4-F-3-Methyl-α-PVP) hydrochloride without reference substance.@*METHODS@#The direct-injection electron ionization-mass spectrometry (EI-MS), GC-MS, electrospray ionization-high resolution mass spectrometry (ESI-HRMS), ultra-high performance liquid chromatography-high resolution tandem mass spectrometry (UPLC-HRMS/MS), nuclear magnetic resonance (NMR), ion chromatography and Fourier transform infrared spectroscopy (FTIR) were integrated utilized to achieve the structural analysis and characterization of the unknown compound in the sample, and the cleavage mechanism of the fragment ions was deduced by EI-MS and UPLC-HRMS/MS.@*RESULTS@#By analyzing the direct-injection EI-MS, GC-MS, ESI-HRMS and UPLC-HRMS/MS of the compound in the samples, it was concluded that the unknown compound was a structural analog of 4-F-α-PVP, possibly with one more methyl group in the benzene ring. According to the analysis results of 1H-NMR and 13C-NMR, it was further proved that the methyl group is located at the 3-position of the benzene ring. Since the actual number of hydrogen in 1H-NMR analysis was one more than 4-F-3-Methyl-α-PVP neutral molecule, it was inferred that the compound existed in the form of salt. Ion chromatography analysis results showed that the compound contained chlorine anion (content 11.14%-11.16%), with the structural analysis of main functional group information by FTIR, the unknown compound was finally determined to be 4-F-3-Methyl-α-PVP hydrochloride.@*CONCLUSIONS@#A comprehensive method using EI-MS, GC-MS, ESI-HRMS, UPLC-HRMS/MS, NMR, ion chromatography and FTIR to identify 4-F-3-Methyl-α-PVP hydrochloride in samples is established, which will be helpful for the forensic science laboratory to identify this compound or other analog compounds.

Impairment of superovulation on the establishment of pregnancy in mice / 解剖学报

Objective To explore whether superovulation impairs the process of pregnancy establishment in mice by changing the intrauterine environment. Methods The implantation and pregnancy of superovulated and normal mice were compared. The superovulated mice were subjected to unilateral tubal ligation on day 0. 5 and blastocysts were transplanted to the other uterine horn on day 2. 5. The number of implantation sites of bilateral uterine horn was compared. The differences between preimplantation uteri of superovulated and normal pseudopregnancy mice were compared by tissue sections and high-throughput sequencing. Bioinformatics analysis was performed on the differentially expressed genes in two groups. Results Compared with the control group, the pregnancy rate of mice in the superovulation group decreased significantly. The number of implantation sites in the superovulation group was higher than the control. There was no significant difference in the pregnancy rate of the uterine horn between the control side and the transplanted side of the superovulated mice. The endometrium was thinned and the number of glands was reduced in superovulated pseudopregnancy mice. The gene expression patterns of preimplantation uterus in superovulation pseudopregnancy and normal pseudopregnancy mice were different. There were 1097 significantly differentially expressed genes, including 752 up-regulated genes and 345 down-regulated genes. Bioinformatics analysis showed that differentially expressed genes are mainly involved in biological processes, such as decidualization, response to progesterone, positive regulation of angiogenesis. They were mainly enriched in FoxO signaling pathway, cell cycle pathway and steroid biosynthesis pathway. Conclusion Superovulation impaired the process of establishing pregnancy and altered the gene expression patterns of biomarker of uterine receptivity in mice.

Decidualization induced by mouse tetraploid embryos / 解剖学报

Objective To investigate the role of inner cell mass (ICM) in decidualization using decidua induced by two-cell embryos or tetraploid embryos through tubal embryo transfer. Methods Tetraploid embryo, as the inner cell mass-deficient embryo, were produced by electrofusion of mouse 2-cell embryos. Decidua was induced by 2-cell embryos or tetraploid embryos through tubal embryo transfer. Decidua induced by 2-cell embryos was employed as a control. Morphologic and implantation site of decidua were compared between two-cell embryo-induced decidua and tetraploid embryo-induced decidua. The differentially expressed microRNA ( miRNA ) was screened by high-throughput sequencing. The target genes of differentially expressed miRNA in two groups were screened for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results Tetraploid embryo-induced decidual tissue and 2-cell embryo- induced decidual tissue were very similar at the implantation site, but there were significant differences in decidual morphology. There were 16 miRNAs differentially expressed in decidua of the two groups, of which 11 miRNAs (miR-466f- 3p, miR-302 d-3p, miR-466i-5p, miR-465c-5p, miR-302a-5p, miR-7068-3p, miR-741-3p, miR-302a-3p, miR-433-5p, miR-144-5p, miR-878-5p) were up-regulated in tetraploid embryo-induced decidua and 5 miRNAs (miR-690, miR-193b- 5p, miR-147-3p, novel_327, miR-363-3p) were down-regulated in tetraploid embryo-induced decidua. Bioinformatics analysis showed that the target genes played function such as protein binding and ion binding, and mainly involved in cyclic guanosine monophosphate-protein kinase G (cGMP-PKG) signaling pathway, estrogen signaling pathway, vascular endothelial growth factor (VEGF) signaling pathway and so on. Conclusion ICM plays an important role in decidualization.

Culture of HSV-2 and cloning of specific fragment of the gG-2 gene / 中华男科学杂志

<p><b>OBJECTIVE</b>To clone the glycoprotein G gene and its specific fragment with high conservation and antigenicity by culturing and amplifying herpes simplex virus type 2 and extracting its whole genome.</p><p><b>METHODS</b>We obtained a great deal of suspension with HSV-2 virus after infecting the cultured Hela cells with HSV-2 virus, extracted the whole genome of the virus by the phenol-chloroform method, and amplified the US4 gene coding gG-2 by PCR. Then we selected the specific target fragment according to the amino acid sequence alignment of the gG-2 gene and cloned it with the designed primers with restricted endonuclease sites.</p><p><b>RESULTS</b>We successfully obtained a lot of suspension with HSV-2 virus, and cloned the gG-2 gene from the whole genuine and its specific target fragment. Sequencing showed that both the sequences were identical with those printed in the GenBank.</p><p><b>CONCLUSION</b>It is feasible to obtain the virus genome and specific fragment of the gG-2 gene from virus-infected cells, especially for HSV-2 virus with relatively stable hereditary trait. It has prepared the ground for further constructing the expression plasmid of the specific fragment, expressing related proteins and identifying their antigenicity.</p>

Construction of plasmid expression vector for specific peptide of the rubella virus E1 gene / 中华男科学杂志

<p><b>OBJECTIVE</b>To construct a recombinant plasmid vector of the RV specific fragment for expressing the specific fragment of RV E1 protein.</p><p><b>METHODS</b>RNA of the RV attenuated live vaccine Wistar RA27/3 strain was extracted and reversely transcribed. The specific fragment of the E1 gene was amplified and the PCR products cloned in the vector pGEX-2T after purification. Positive clones were selected and identified by two-enzyme digestion and sequence analysis.</p><p><b>RESULTS</b>A 330 bp target fragment was successfully cloned, and the sequence of the recombinant plasmid was consistent with the original sequence.</p><p><b>CONCLUSION</b>Successful cloning of the RV El specific fragment and the construction of the recombinant plasmid have laid a foundation for further expressing the recombinant protein.</p>

Vardenafil for refractory erectile dysfunction: the latest advances / 中华男科学杂志

In recent years, more and more attention has been drawn to the role of phosphodiesterase 5 (PDE5) in penile erection. The cyclic nucleotide (cGMP) signaling pathway mediates the smooth-muscle relaxing effect of nitric oxide necessary for normal erectile function. Down-regulation of this pathway is the pathological pivot of many forms of erectile dysfunction (ED) and leads to the development of some chronic diseases. Therapeutic outcomes have shown that vardenafil is effective and safe in the treatment of ED associated with dyslipidemia, hypertension, depression, diabetes, radical retropubic prostatectomy, spinal cord injury, sildenafil failure, renal transplantation, chronic prostatitis and that accompanied by premature ejaculation. Vardenafil provides a reasonable therapeutic alternative for these refractory ED patients. In addition, vardenafil can prolong erectile duration of ED patients.

L-carnitine improves the reproductive function of male rats with ornidazole-induced asthenospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the protective effect of L-carnitine (LC) on the reproductive function of male rats with asthenospermia induced by ornidazole (ORN).</p><p><b>METHODS</b>Forty male SD rats (200-230 g) were randomly divided into Groups A (control: 0.5% carboxymethylcellulose solution), B (medium-dose ORN: 400 mg/kg/d), C (medium-dose ORN + LC: ORN 400 mg/kg/d + LC 100 mg/kg/d), D (high-dose ORN: 800 mg/kg/d), and E (high-dose ORN + LC: ORN 800 mg/kg/d + LC 100 mg/kg/d). All the rats were treated via gastric gavage for 20 days consecutively, and then killed for the detection of sperm motility and the sperm count of the cauda epididymis.</p><p><b>RESULTS</b>Compared with Group A, there was a significant decrease in sperm motility and sperm count in Groups B and D (P < 0.05), while Group C showed a significant increase in both the parameters as compared with B (P < 0.05), but with no significant difference from A (P > 0.05). Group E exhibited no obvious improvement in sperm motility and sperm count, with no difference from D (P > 0.05).</p><p><b>CONCLUSION</b>L-carnitine can improve the sperm motility and sperm count of the male rats with ornidazole-induced asthenospermia.</p>

The safety of tadalafil in the treatment of erectile dysfunction / 中华男科学杂志

The cyclic nucleotide (cGMP) signalling pathway mediates the smooth-muscle relaxing effects of nitric oxide necessary for normal erectile function. Down-regulation of this pathway is the pathophysiological pivot of many forms of erectile dysfunction (ED) and leads to the development of some chronic diseases, such as hypertension and type 2 diabetes mellitus. Therefore, selective inhibition of the enzyme that catalyses the degradation of cGMP promotes erectile responses to sexual stimulation. Recently, a new phosphodiesterase type 5 (PDE-5) inhibitor tadalafil has emerged, which has a prolonged half-life. Here is a review of recent studies on the safety of tadalafil in the treatment of ED.

Identification of the fumatory Radix Bupleuri with sulfur by static headspace GC-MS / 中国中药杂志

<p><b>OBJECTIVE</b>To identify whether Radix Bupleuri (Bupleurum chinense) was fumed with sulfur.</p><p><b>METHOD</b>A static headspace GC-MS method was used to detect sulfur in the fumatory Radix Bupleuri, the authentic samples free of sulfur was detected as reference.</p><p><b>RESULT</b>Sulfur was detected in six samples from nine samples collected in different locations.</p><p><b>CONCLUSION</b>The method can be used to detect sulfur rapidly in the fumatory Radix Bupleuri with sulfur.</p>