An ideal animal model of azoospermia would be a powerful tool for the evaluation of spermatogonial stem cell (SSC) transplantation. Busulfan has been commonly used to develop such a model, but 30%-87% of mice die when administered an intraperitoneal injection of 40 mg kg^-1. In the present study, hematoxylin and eosin staining, Western blot, immunofluorescence, and quantitative real-time polymerase chain reaction were used to test the effects of busulfan exposure in a mouse model that received two intraperitoneal injections of busulfan at a 3-h interval at different doses (20, 30, and 40 mg kg^-1) on day 36 or a dose of 40 mg kg^-1 at different time points (0, 9, 18, 27, 36, and 63 days). The survival rate of the mice was 100%. When the mice were treated with 40 mg kg^-1 busulfan, dramatic SSC depletion occurred 18 days later and all of the germ cells were cleared by day 36. In addition, the gene expressions of glial cell line-derived neurotrophic factor (GDNF), fibroblast growth factor 2 (FGF2), chemokine (C-X-C Motif) ligand 12 (CXCL12), and colony-stimulating factor 1 (CSF1) were moderately increased by day 36. A 63-day, long-term observation showed the rare restoration of endogenous germ cells in the testes, suggesting that the potential period for SSC transplantation was between day 36 and day 63. Our results demonstrate that the administration of two intraperitoneal injections of busulfan (40 mg kg^-1 in total) at a 3-h interval to mice provided a nonlethal and efficient method for recipient preparation in SSC transplantation and could improve treatments for infertility and the understanding of chemotherapy-induced gonadotoxicity.
Adult Germline Stem Cells/transplantation* ; Animals ; Azoospermia/chemically induced* ; Busulfan/toxicity* ; Disease Models, Animal ; Infertility, Male/chemically induced* ; Injections, Intraperitoneal ; Male ; Mice ; Spermatogenesis/drug effects* ; Spermatogonia/drug effects* ; Stem Cell Transplantation/methods*

Objective To understand the relationship between the concentration of air pollutants and daily emergency department visits for different diseases (circulatory system disease, digestive system disease, nervous system disease and respiratory system disease) in Guangzhou, Guangdong Province. Methods The daily average concentrations of sulfur dioxide (SO2), nitrogen dioxide (NO2, carbon monoxide (CO) and PM2.5 and the daily maximum 8-hour concentrations of O3, the daily average temperature, the relative humidity and cause -specific emergency department visits of the four major diseases from 2015 to 2017 were collected in Guangzhou. Semi-parametric generalized additive model was used to analyze the relationship between the concentration of pollutants and daily cause-specific emergency department visits. Results The daily average concentrations of SO2, NO2, CO, O3 and PM2.5 during the study period were 13.24 μg /m3, 45.96 μg /m3, 0.97 mg /m3, 123.77 μg /m3 and 36.22 μg /m3, respectively. For circulatory system disease，the independently significant associations of SO2 with emergency department visits in single-pollutant models (2.91%, 95% CI: 1.00%-4.85%), and multipollutant models (4.39%, 95% CI: 1.22%-7.67%) were observed. Conclusion The ambient SO2 increases the risk of emergency department visits due to circulatory diseases in Guangzhou. Comprehensive prevention and control measures should be taken to reduce the emission of SO2.

Objective To investigate the uptake mechanism of HepG2.2.15 cells to the nanoparticles co-loaded with syringopicroside and hydroxytyrosol (SH-NPs). Methods The nanoparticles were prepared by using a nanoprecipitation method with mPEG-PLGA as nano-carrier co-loaded with syringopicroside and hydroxytyrosol. The uptake mechanism of HepG2.2.15 cells to SH-NPs was studied by fluorescence microscopy and flow cytometry using fluoresceineisothiocyanate (FITC) as a fluorescent marker. Results With colchicine as the inhibitor, the incubation time ranged from 0.5 to 24 h, the percentage of positive cells increased from 1.9% to 56.4%; When the drug concentration was 125, 250 μg/mL and 500 μg/mL, the positive cell percentages were 4.9%, 3.4% and 3.9%. With chloroquine as the inhibitor; the incubation time ranged from 0.5 to 24 h, the percentage of positive cells increased from 7.4% to 55.4%; When the drug concentration was 125, 250 and 500 μg/mL, the percentage of positive cells was 19.5%, 22.5% and 27.6%. Conclusion Colchicine and chloroquine have an inhibitory effect on HepG2.2.15 cells uptake, and the uptake of SH-NPs in HepG2.2.15 cells was positively correlated with drug concentration and incubation time. It can be concluded that the uptake mechanism of HepG2.2.15 cells to SH-NPs was nonspecific adsorption endocytosis.

Two previously undescribed steroidal compounds, 16, 23-epoxy-22, 26-epimino-cholest-22(N), 23, 25(26)-trien-3β-ol-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside (1) and 26-O-β-D-glucopyranosyl-(25R)-5α-furost-20(22)-en-3β, 26-diol (2), together with 7 known ones including 26-O-β-D-glucopyranosyl-(25R)-5, 20(22)-dien-furost-3β, 26-diol (3), (25R)-5-en-spirost-3β-ol-O-β-D-glucopyranosyl-(1→4)-[α-L-rhmanopyranosyl-(1→2)]-β-D-galactopyranoside (4), funkioside D (5), aspidistrin (6), tigogenin-3-O-β-D-lucotrioside (7), desglucolanatigonin II (8), and degalactotigonin (9), were isolated from Solanum lyratum Thunb. Their cytotoxic activities were tested in two cancer cell lines by MTT method. One of the steroidal glycosides (6) showed significant cytotoxic activity against gastric cancer SGC7901 and liver cancer BEL-7402 cells.
Alkaloids ; chemistry ; isolation & purification ; toxicity ; Antineoplastic Agents ; chemistry ; isolation & purification ; toxicity ; Cell Line, Tumor ; Cell Survival ; drug effects ; Glycosides ; chemistry ; pharmacology ; toxicity ; Humans ; Inhibitory Concentration 50 ; Molecular Structure ; Phytosterols ; chemistry ; isolation & purification ; toxicity ; Plant Extracts ; chemistry ; toxicity ; Plants, Medicinal ; chemistry ; Solanum ; chemistry ; Sterols ; chemistry ; pharmacology ; toxicity

OBJECTIVE: To study whether tricalcium phosphate(TCP) wear particles cause injuries of periprosthetic osteocytes in the mouse calvaria, and to explain its molecular mechanism. METHODS: Thirty six-week(ICR)male mice were randomly divided into sham group, model (TCP) group and 3-methyladenine (3-MA) group. A murine calvarial model of osteolysis was established by 30 mg of TCP wear particles implantation over the periosteum around the middle suture of calvaria in mice. On the second postoperative day, the autophagy specific inhibitor 3-MA (1.0 mg/kg) was subcutaneously injected to the calvaria in the 3-MA-treated mice every other day. After 2 weeks, blood and the calvaria were obtained. Micro-CT was used to detect bone mineral density(BMD), bone volume fraction (BVF) and porosity number. HE staining and flow cytometry were performed to analyze the viability and apoptosis of periprosthetic osteocytes. The serum levels of dentin matrix protein 1(DMP-1) and sclerostin (SOST) were determined by ELISA. The proteins expressions of DMP-1, SOST, Beclin-1 and microtuble-associated protein 1 light chain 3 (LC-3) were detected by Western blot in the calvaria osteocytes. RESULTS: Compared with the sham group, the mice in the TCP group showed that a significant decrease in the viability of periprosthetic osteocytes, but obvious increases in number of osteocytes death and osteocytes apoptosis (<0.05), and in serum level and protein expression of SOST; significant decreases in serum level and protein expression of DMP-1 (<0.05), and remarkable up-regulation of autophagy-related factors beclin-1 and the conversion of LC3-Ⅱ from LC3-I in the calvaria osteocytes. Compared with TCP group, the mice in the 3-MA group showed that injuries of calvaria osteocytes were obviously aggravated, and osteocytes apoptosis was significantly increased (<0.05). CONCLUSIONS TCP wear particles can cause injuries of periprosthetic osteocytes via activation of apoptosis and autophagy, which promotes osteolysis around the prosthesis osteolysis and joint aseptic loosening.
Animals ; Apoptosis ; Beclin-1 ; metabolism ; Bone Density ; Calcium Phosphates ; adverse effects ; Extracellular Matrix Proteins ; metabolism ; Glycoproteins ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Microtubule-Associated Proteins ; metabolism ; Osteocytes ; pathology ; Osteolysis ; Prostheses and Implants ; adverse effects ; Skull

This study was aimed to explore the feasibility of identification of traditional Chinese medicine (TCM) constitution based on photoplethysmography (PPG) frequency domain analysis,and to establish an automatic identification system (AIS) of TCM constitution with simple operation and good generalization.The Classification and Determination of Constitution in TCM was used to evaluate the subjects' constitutional state.Pulse wave sensor was used to collect the pulse wave signal.Spectral characteristics were analyzed by power spectrum analysis.Eight classification algorithms of data mining were used to establish the model of qi-insufficiency constitution.The results showed that the pulse wave harmonic peak was that qi-insufficiency constitution was higher than health group;the model was optimal when random forest algorithm was used;and the accuracy of 10-CV test was 80.4％.Sensitivity reached 73.2％.Specificity and receiver operator characteristic were all above 0.8.The model has certain reliability.It was concluded that it was feasible to identify qiinsufficiency constitution by means of frequency domain analysis of PPG.It provides preliminary work for the construction of AIS of TCM constitution as well as a simple and quick assistant tool for the constitution identification of subhealth group in family health care.

Objective To evaluation in vitro activity and synergy of Yin-hua Miyan Ling against main clinical isolated strains.Methods All of 285 stains from 18 tertiary hospitals in recent three years were studied. Minimal inhibition concentrations ( MIC ) were tested by Agar dilution method and combination effects were measured by Checkerboard method. Results Yinhua Miyan Ling showed more in vitro activity against Sta-phylococcus and Proteus mirabilis, MIC values from 3.13~6.25 mg? mL-1 . On the other hand, Yinhua Miyan Ling showed the same antibiotic activi-ty against extended-spectrumβlactamases ( ESBLs) positive and nega-tive strains or methecillin susceptible and resistant Staphylococcus.Syner-gy was detected with Yinhua Miyan Ling in combination with amoxillin/clavulanate, cefuroxime and levofloxacin against gram-positive strains, especially methicillin-resistant Staphylococcus aureus ( MRSA).Against E.faecalis, synergy was observed in Yinhua Miyan Ling and cefuroxime. Additive effect was found between Yinhua Miyan Ling and amoxillin/clavulanate against 100%E.coli.Conclusion Synergy was found with Yinhua Miyan Ling in combination withβlactams and quinolones against gram -positive strains and additive effect was found against gram-negative bacilli.

Objective To understand the resistance, molecular charac-teristics of oxacillin susceptible methicillin-resistant Staphylococcus au-reus ( OS-MRSA) strains isolated 2011-2012 in China.Methods A total of 297 methicillin -resistant Staphylococcus aureus ( MRSA ) were collected in the Ministry of Health National Antimicrobial Resistance Sur-veillance Net program and the susceptibility test were performed by agar dilution method.All the oxacillin-susceptible MRSA strains were con-firmed to be Staphylococcus aureus by detecting nuc gene.The molecular typing methods included SCCmec and spa typing.Results A total of 14 isolates ( 4.9%) were oxacillin susceptible methicillin-resistant Staphy-lococcus aureus among 288 MRSA isolates and most of them were from se-cretion (50%, 7/14) and blood (28.6%, 4/14).The susceptibility rate to gentamicin, amikacin, rifampicin and trimethroprim/sulfamethox-azole was more than 90% and the rate to quinolones, tetracycline and chloramphenicol was about 80%, 28.6%, 28.6%, respectively.Moreo-ver, the resistance rate to erythromycin, azithromycin and clindamycin was more than 90%.PCR revealed that 14 isolates were Staphylococcus aureus and mecA positive and the most predominant clone was SCCmecⅤ-spat437 (42.9%,6/14).Conclusion The most predominant clone of the OS-MRSA isolates in China was SCCmecⅤ-spat437; most of OS-MRSA isolates were susceptible to the majority of antibacterial agents except macrolides, tetracycline and chloramphenicol.

Objective To evaluate the antibacterial activities of severalβ-lactam/β-lactam inhibitor combinations in vitro.Methods All the clinical isolates were Enterobacteriaceae collected from hospitals and the susceptibility tests were performed using agar dilution method recom-mended by CLSI.The minimum inhibitory concentration ( MIC ) results were interpreted according to the breakpoints of CLSI (2014) guideline.The data was analyzed by software SPSS17.0.Results A total of 2166 Enterobacteriaceae were collected and were performed MIC tests.Based on the MIC results, the MIC50 and MIC90 of piperacillin to Escherichia coli were 256 , 512 μg · mL-1 , while piperacillin/tazobactam were 2 , 8μg· mL-1 , respectively.The sensitive rates of piperacillin/tazobactam to ESBLs -producing Escherichia coli and Klebsiella pneumoniae were 93.7% and 86.1%, cefoperazone/tazobactam were 91.5% and 82.5%, while cefoperazone /sulbactam were 74.4% and 58.39%. Among ESBLs -non -producing Escherichia coli and Klebsiella pneumoniae, there is not much difference between using β-lactam/β-lactam inhibitor combinations and β-lactams alone.β-lactam antibiotics have higher sensitive rates to Proteus vulgaris than Proteus mirabilis, butβ-lactam/β-lactam inhibitor combinations have no difference against them.The sensitive rate of mezlocillin/sulbactam to Morganella spp.was 97.4%, significantly higher than mezlocillin alone with sensitive rate 76.3%.The MIC90 of cefotaxime/tazobactam and ceftriaxone/tazobactam were lower than using cefotaxime and ceftriaxone alone.Conclusion β-lactam/β-lactam inhibitor combinations have greater antimicrobial activity to Enterobacteriaceae than β-lactams alone, especially for ESBLs-producing Escherichia coli and Klebsiella pneumoniae.Among them, piperacillin/tazobactam has the greatest antimicrobial activity.

Adolescent ; Adult ; Animals ; Bites and Stings ; Dog Diseases ; virology ; Dogs ; Female ; Humans ; Male ; Middle Aged ; Nucleocapsid Proteins ; genetics ; Phylogeny ; Post-Exposure Prophylaxis ; Rabies ; prevention & control ; veterinary ; virology ; Rabies Vaccines ; immunology ; Young Adult