[Lycium barbarum miR2911-loaded exosomes promote spermatogenic function recovery in rats with non-obstructive azoospermia by regulating Wnt/ß-catenin signaling pathways].

OBJECTIVE: To investigate the effect of exosomes loaded with Lycium barbarum miRNA (Lb-miR2911) on spermatogenic function recovery in non-obstructive azoospermia (NOA) rats through cross-regulation of the Wnt/ß-catenin signaling pathways. METHODS: We established an NOA model in 30 four-week-old male SD rats by intraperitoneal injection of busulfan. At 5 weeks after modeling, we equally randomized the rats into a model control group (MC，untreated), an Lb-miR2911EXO group (Lb-miR2911EXO ，treated by intratesticular injection of Lb-miR2911-loaded exosomes), and a sham group (Shame，treated by intratesticular injection of exosomes-empty drug), with another 10 male SD rats taken as normal controls（NC）. We observed the uptake and metabolic changes of Lb-miR2911 in the testis tissue of the rats by RNA FISH at 2 and 6 weeks after treatment, detected cell proliferation, spermatogenesis and gene expressions of the Wnt/ß-catenin signaling pathways in the testis tissue by Transcriptome sequencing analysis combined with Western blot and RT-PCR at 12 weeks, evaluated the recovery of the spermatogenic function based on the testis tissue morphology and sperm quality, and assessed the organ toxicity of Lb-miR2911 in the tissue and organs of the rats based on histomorphological analysis and the levels of serum TNF-α, IL-1ß, Aspartate aminotransferase (AST), Alanine aminotransferase (ALT) and other relevant indicators. RESULTS: After 12 weeks of treatment, histomorphological analysis showed regular arrangement of spermatogenic cells at all levels in the testis tissue, with a large number of mature sperm in the tubular lumen, and with significantly higher Johnsen scores, testis weight, testicular index, sperm concentration and sperm motility in the Lb-miR2911EXO than in the sham group (all P< 0.05). Compared with the model controls, the Lb-miR2911EXO group exhibited remarkably down-regulated gene expression of DACT3 (P< 0.05), up-regulated expressions of DVL2 and ß-catenin (P< 0.05), elevated levels of p-DVL2 and ß-catenin (nucleus) proteins (P< 0.05), increased expressions of cell proliferation-related genes CCND1, CCNE1 and CCNE2 (P< 0.05) and spermatogenesis-related genes DMC1, CCR6, JAM2 and KLC3 (P< 0.05). No pathological changes were observed in the lung, liver and kidney tissues of the rats, or in the levels of serum TNF-α, IL-1ß, AST, ALT, creatinine and urea nitrogen in the rats treated with Lb-miR2911EXO compared with the normal controls (P > 0.05). CONCLUSION: Lb-miR2911-loaded exosomes promote spermatogenic function recovery in NOA rats through cross-regulation of the DACT3, Wnt and ß-catenin signaling pathways.

Metabolic profile of salidroside in rats using high-performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry.

A high-performance liquid chromatography coupled to Fourier transform ion cyclotron resonance mass spectrometry (HPLC-FT-ICR MS) method was developed to study the in vivo metabolism of salidroside for the first time. Plasma, urine, bile, and feces samples were collected from male rats after a single intragastric gavage of salidroside at a dose of 50 mg/kg. Besides the parent drug, a total of seven metabolites (three phase I and four phase II metabolites) were detected and tentatively identified by comparing their mass spectrometry profiles with those of salidroside. Results indicated that metabolic pathways of salidroside in male rats included hydroxylation, dehydrogenation, glucuronidation, and sulfate conjugation. Among them, glucuronidation and sulfate conjugation were the major metabolic reactions. And most important, the detection of the sulfation metabolite of p-tyrosol provides a clue for whether the deglycosylation of salidroside occurs in vivo after intragastric gavage. In summary, results obtained in this study may contribute to the better understanding of the safety and mechanism of action of salidroside.

Two new indole-diterpenoids from the fungus Penicilliumcrustosum YN-HT-15.

Two new indole-diterpenoids named penijanthine B and penitrem H were isolated from the metabolites of the fungus Penicilliumcrustosum YN-HT-15, which was isolated from the red soil of Yunnan Province of China. The structures were determined on spectroscopic analyses and CD spectra.

Causes of death after hip fracture in senile patients.

Hip trauma has been a leading cause of death in senile patients for more than a centenary. Although the mortality decreased due to the advanced technique in medication, surgery and nursing, the increasing mortality should not be neglected in elders after orthopedic operation nowadays. Many factors are considered to influence the causes of death after trauma, such as age, gender, personal customs, comorbidities, types of fracture, timing of surgery, procedure, anesthesia, complications, medical treatment, activity of daily living, or even marriage status. This article reviews these causes from the aspects of patient's own factors, iatrogenic factors, medical treatment and other factors and provides some clues for further clinical application according to the recent foreign and domestic researches. According to the present research, it is essential for surgeons to perform a comprehensive estimation for patients suffering from hip trauma.

Analysis of the fatty acid from Bupleurum Chinense DC in China by GC-MS and GC-FID.

Fatty acid of the root of the medicinal plant of Bupleurum Chinense DC in China has been investigated by gas chromatography combined with mass spectroscopy. After methyl-esterification, eight fatty acid compositions were identified by GC-MS. A simple and rapid determination of the fatty acid has been firstly developed by GC-FID. The derivatization condition was investigated in order to validate this method. Palmitic acid, palmitoleic acid, oleic acid and linoleic acid were analyzed simultaneously by internal standard method. The validity of method has been examined both experimentally with good recovery, intra-assay precisions and linearity. The quick and accurate method of capillary gas chromatography could be used for the analysis of the fatty acid from Bupleurum Chinense DC.

Two new monoterpenoid glycosides from Mentha spicata L.

Two new monoterpenoid glycosides, spicatoside A and spicatoside B, were isolated from the whole herbs of Mentha spicata L. which have anti-inflammatory and hemostatic activities. Their structures have been determined on the basis of spectral and chemical analysis. They are (+)-5-[1-(beta-D-glucopyranosyloxymethyl)ethenyl]-2-methyl-2-cyclohexen-1-one (1), and (-)-5-[[2-(beta-D-glucopyranosyloxy)-1-hydroxy-l-methyl]ethyl]-2-methyl-2-cyclohexen-1-one (2).