ObjectiveBased on ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap-linear ion-trap mass spectrometry（UPLC-Orbitrap Fusion Lumos Tribrid-MS）， the plasma concentration of ziyuglycoside Ⅰ was determined at different time points after oral administration， and its pharmacokinetic characteristics in normal rats and rats with acute kidney injury were compared. MethodsRats were randomly divided into normal group and model group， the model group received intraperitoneal cisplatin（10 mg·kg^-1） to establish the acute kidney injury model， the normal group was given the same volume of saline. After successful modeling， rats in the normal and model groups were randomly divided into the normal low， medium and high dose groups（2.5， 5， 7.5 mg·kg^-1） and the model low， medium and high dose groups（2.5， 5， 7.5 mg·kg^-1）， 6 rats in each group， and the plasma was collected at different time points after receiving the corresponding dose of ziyuglycoside Ⅰ. Then， the concentration of ziyuglycoside Ⅰ in rat plasma was determined by UPLC-Orbitrap Fusion Lumos Tribrid-MS， and the drug-time curve was poltted. The pharmacokinetic parameters were calculated by Kinetica 5.1 software， and the differences in pharmacokinetic parameters between different administration groups were compared by independent sample t-test with SPSS 22.0. ResultsThe pharmacokinetic results showed that after receiving the different doses of ziyuglycoside Ⅰ， its concentration increased first and then decreased， and all of them reached the maximum plasma concentration at about 0.5 h. The area under the curve（AUC_0-t） and mean retention time（MRT_0-t） of normal and model rats increased with the increased dose， and the clearance（CL） decreased with the increasing dose. Compared with the normal group， the AUC_0-t was significantly increased（P<0.01）， peak concentration（C_max） and CL decreased in model rats at different doses， indicating that the physiological state of the rats could affect the absorption and elimination of ziyuglycoside Ⅰ in vivo. ConclusionThe pharmacokinetic characteristics of ziyuglycoside Ⅰ are quite different in normal rats and acute kidney injury model rats， which may be due to the change of the body environment in the pathological state， then lead to changes in absorption and metabolic processes.

BACKGROUND:Psoralen has a strong anti-osteoporotic activity and may have a restorative effect on chemotherapy-induced osteoporosis. OBJECTIVE:To explore the restorative effect of psoralen on the osteogenic differentiation of bone marrow mesenchymal stem cells in mice inhibited by cyclophosphamide and its mechanism. METHODS:C57BL/6 mouse bone marrow mesenchymal stem cells were isolated and cultured.Effect of psoralen on viability of bone marrow mesenchymal stem cells was detected by MTT assay.Osteogenic induction combined with alkaline phosphatase staining was used to determine the optimal dose of psoralen to restore the osteogenic differentiation of bone marrow mesenchymal stem cells inhibited by cyclophosphamide.The mRNA expression levels of Runx2,alkaline phosphatase,Osteocalcin,osteoprotegerin,and Wnt/β-catenin signaling pathway-related genes Wnt1,Wnt4,Wnt10b,β-catenin,and c-MYC were measured by RT-qPCR at different time points under the intervention with psoralen.The protein expression of osteogenic specific transcription factor Runx2 and Wnt/β-catenin signaling pathway related genes Active β-catenin,DKK1,c-MYC,and Cyclin D1 was determined by western blot assay at different time points under the intervention with psoralen. RESULTS AND CONCLUSION:(1)There was no significant effect of different concentrations of psoralen on the viability of bone marrow mesenchymal stem cells.The best recovery of the inhibition of osteogenic differentiation of bone marrow mesenchymal stem cells caused by cyclophosphamide was under the intervention of psoralen at a concentration of 200 μmol/L.(2)Psoralen reversed the reduction in osteogenic differentiation marker genes Runx2,alkaline phosphatase,Osteocalcin and osteoprotegerin mRNA expression and Runx2 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(3)Psoralen reversed the decrease in Wnt/β-catenin pathway-related genes Wnt4,β-catenin,c-MYC mRNA and Active β-catenin,c-MYC,and Cyclin D1 protein expression and the increase in DKK1 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(4)The results showed that cyclophosphamide inhibited osteogenic differentiation of bone marrow mesenchymal stem cells in mice,and psoralen had a restorative effect on it.The best intervention effect was achieved at a concentration of 200 μmol/L psoralen,and this protective effect might be related to the activation of Wnt4/β-catenin signaling pathway by psoralen.

Objective To investigate the shedding of CAV2-ΔE3-CGS after immunization and the background of canine adenovirus (CAV) infection, and to establish a dual fluorescent quantitative PCR detection method for rabies virus (RV) and canine adenovirus type 2 (CAV2). Methods A dual fluorescent quantitative PCR detection method was established by designing specific primers and probes for E1 gene of CAV and G gene of RV for the detection of CAV2-ΔE3-CGS. Oral swabs, anal swabs and environmental samples of stray dogs from experimental animal farm and dog detention center were tested. Results The standard curves generated by this method were Y=-3.351 × logX + 44.895, R² = .999 and Y=-3.413 × logX + 45.192, R²=0.996, respectively. The linear relationships were good, and the minimum detection limits were both 102 copies/μL. CAV2-ΔE3-CGS was not detected in experimental animal farm. CAV was detected in dog detention center, and the positive rates were 5.88% (5/85) in oral swabs, 8.24% (7/85) in anal swabs, and 4% (1/25) in environmental samples. Conclusion The dual fluorescent quantitative PCR method can be used for the detection of CAV2-ΔE3-CGS after immunization and the investigation of CAV infection. The present study has shown that no CAV2-ΔE3-CGS has been detected after immunization and CAV infection rate of stay dogs is low in Shanghai. CAV2-ΔE3-CGS oral immunization meets requirement and is applicable.

In order to determine the changes of bacterial community structure and function in the early, middle and late stage of aerobic composting of chicken manure, high-throughput sequencing and bioinformatics methods were used to determine and analyze the 16S rRNA sequence of samples at different stages of composting. Wayne analysis showed that most of the bacterial OTUs in the three composting stages were the same, and only about 10% of the operational taxonomic units (OTUs) showed stage specificity. The diversity indexes including Ace, Chao1 and Simpson showed a trend of increasing at first, followed by decreasing. However, there was no significant difference among different composting stages (P < 0.05). The dominant bacteria groups in three composting stages were analyzed at the phylum and genus levels. The dominant bacteria phyla at three composting stages were the same, but the abundances were different. LEfSe (line discriminant analysis (LDA) effect size) method was used to analyze the bacterial biological markers with statistical differences among three stages of composting. From the phylum to genus level, there were 49 markers with significant differences among different groups. The markers included 12 species, 13 genera, 12 families, 8 orders, 1 boundary, and 1 phylum. The most biomarkers were detected at early stage while the least biomarkers were detected at late stage. The microbial diversity was analyzed at the functional pathway level. The function diversity was the highest in the early stage of composting. Following the composting, the microbial function was enriched relatively while the diversity decreased. This study provides theoretical support and technical guidance for the regulation of livestock manure aerobic composting process.
Animals ; Manure/microbiology* ; Chickens/genetics* ; Composting ; RNA, Ribosomal, 16S/genetics* ; Soil ; Bacteria/genetics*

Objective:To investigate the clinical, histologic, immunohistochemical (IHC) and molecular genetic features of clear cell carcinoma (CCC) of salivary gland in the head and neck regions.Methods:Seven cases of CCC diagnosed in the Department of Pathology, the First Affiliated Hospital of Nanjing Medical University from 2018 to 2021 were included. The clinical and pathologic data, HE sections and IHC staining were reviewed, and EWSR1 gene translocation was detected by fluorescence in situ hybridization (FISH). The relevant literature was also reviewed.Results:There were five males and two females, with an age range of 32 to 71 years (mean 50 years). The tumors were located in the palate, base of tongue, subglottic, right submaxillary and nasopharynx. Histologically the tumors were composed of sheets, nests, and trabecular of large, monomorphic cells which possessed abundant clear and eosinophilic cytoplasm. The stroma was characterized by abundant hyalinized fibrous strands admixed with cellular fibrous (desmoplastic) tissue. The tumor growth was infiltrative. IHC staining revealed positivity for CKpan and squamous cell immunophenotypic markers (CK5/6, p63 and p40), but negativity for myoepithelial markers (SMA, calponin, GFAP and CD10). The EWSR1 gene translocation was detected by FISH. The prognosis was excellent, with the follow-up periods ranging from 8 months to 33 months. During this period, six patients survived without tumor, only one patient with cervical lymph node metastasis.Conclusions:CCC of salivary gland is rare and needs to be differentiated from various other types of tumors containing clear cells. Awareness of the histopathologic characteristics, and combined with IHC and molecular genetic examination can avoid misdiagnosis. The biological behavior of the tumor is indolent with a good overall prognosis.

Objective:To investigate the effects and mechanism of sodium hydrosulfide on rat epidermal cells intervened with serum from burned rat (hereinafter referred to as burn serum).Methods:The experimental research method was used. Ten male Sprague-Dawley (SD) rats aged eight months were taken to prepare normal rat serum (hereinafter referred to as normal serum), 30 male SD rats aged eight months were taken to prepare burn serum after full-thickness burn, and epidermal cells (the third passage)isolated from 10 SD rats born one day were used for the experiments. The cells were divided into normal serum group treated with normal serum and burn serum group treated with burn serum. Cell counting kit 8 method was used to detect cell survival rate after 1, 2, 4, 6, and 8 h of culture, respectively, to screen the subsequent intervention time of burn serum. The cells were divided into burn serum control group treated only with burn serum and 50, 100, 150, 200, 250 μmol/L sodium hydrosulfide groups treated with burn serum+ sodium hydrosulfide at corresponding final molarity. After 30 min of culture following the burn serum intervention, the cell survival rate was detected as above to screen the subsequent intervention concentration of sodium hydrosulfide. The cells were divided into burn serum control group treated with burn serum only and sodium hydrosulfide only group, glibenclamide only group, and sodium hydrosulfide+ glibenclamide group treated with burn serum+ corresponding reagents. After 5, 10, 15 min of culture following the burn serum intervention, the cell survival rate was detected as above to screen the subsequent intervention time of glibenclamide. The cells were divided into burn serum control group treated with burn serum and sodium hydrosulfide only group, glibenclamide only group, and sodium hydrosulfide+ glibenclamide group treated with burn serum+ corresponding reagents. After completing corresponding culture time of each reagent, the mitochondria were extracted to detect cytochrome c oxidase (CCO) activity using a spectrophotometer, and the protein expression level of adenosine triphosphate (ATP)-sensitive potassium channel was detected by Western blotting. Except for the number of samples for ATP-sensitive potassium channel protein detection, which was 3, the number of samples for the other indicators was 10. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, least significant difference (LSD)- t test, LSD test, and Bonferroni correction. Results:Compared with that of normal serum group, the cell survival rate was significantly decreased in burn serum group after only 4 and 6 h of culture ( t=4.02, 6.42, P<0.05). An overall comparison showed statistically significant differences in cell survival rate among the time points within normal serum group and burn serum group ( F=19.74, 4.48, P<0.05 or P<0.01). Four hours of culture was selected as the subsequent intervention time of burn serum. After 30 min of culture following the burn serum intervention, compared with that of burn serum control group, only 150, 200, 250 μmol/L sodium hydrosulfide groups had a significantly higher cell survival rate ( P<0.01), thus 150 μmol/L was selected as the subsequent intervention concentration of sodium hydrosulfide. Compared with that of burn serum control group, the cell survival rate decreased significantly in glibenclamide only group after 5 and 15 min of culture following burn serum intervention ( P<0.05) and increased significantly in glibenclamide only group after 10 min of culture following the burn serum intervention and sodium hydrosulfide only group at each time point ( P<0.05 or P<0.01). The cell survival rate in sodium hydrosulfide+ glibenclamide group was significantly lower than that of sodium hydrosulfide only group at each time point ( P<0.05). The difference in cell survival rate was statistically significant among the time points within glibenclamide only group ( F=11.81, P<0.01). Five minutes of culture was selected as the subsequent intervention time of glibenclamide. After 35 min of culture following the burn serum intervention, compared with (1.62±0.08) nmol·min -1·mg -1 and 0.682±0.063 in burn serum control group, the CCO activity of cells and the protein expression level of ATP-sensitive potassium channel were significantly increased in sodium hydrosulfide only group ((1.99±0.09) nmol·min -1·mg -1 and 0.932±0.014, P<0.01) and significantly decreased in glibenclamide only group ((1.44±0.09) nmol·min -1·mg -1 and 0.600±0.012, P<0.01); the CCO activity of cells and the protein expression level of ATP-sensitive potassium channel in sodium hydrosulfide+ glibenclamide group ((1.79±0.06) nmol·min -1·mg -1 and 0.744±0.071) was significantly lower than those of sodium hydrosulfide only group ( P<0.05 or P<0.01). Conclusions:Sodium hydrosulfide can improve the survival rate of rat epidermal cells after burn serum intervention, by a mechanism which is related to the alleviation of epidermal cell mitochondrial damage and mediated by ATP-sensitive potassium channel.

OBJECTIVE：To stud y the effects of different processed products of Whitmania pigra on hemorheology and coagulation indexes in acute blood stasis model rats. METHODS ：SD rats were randomly divided into blank group ，model group ， aspirin group ，W. pigra hang-dried product low- ，medium- and high-dose groups ，W. pigra talcum powder-ironed product low- ， medium- and high-dose groups ，W. pigra wine bran-processed product low- ，medium- and high-dose groups ，with 6 rats in each group. Except for blank group ，other groups received subcutaneous injection of epinephrine hydrochloride and ice water bath for 15 d to induce acute blood stasis model. From the 8th day of modeling ，rats in aspirin group were given aspirin 0.2 g/kg intragastrically. Rats in each dose group of W. pigra processed products were given relevant medicine 0.35，1.4，3.5 g/kg intragastrically（calculated by crude drug ）. Rats in blank group and model group were given constant volume of normal saline intragastrically， once a day ， for consecutive 8 days. Hemorheology indexes as whole blood viscosity （high， medium and low shearrate ），plasma viscosity ，erythrocyte com deformation index ，erythrocyte aggregation index ，hematocrit， and blood coagulation indexes as prothrombin time （PT）， mail：wcl19960125@163.com activated partial prothrombin time （APTT），thrombin time （TT）were determined. RESU LTS：Compared with blank group ，whole blood viscosity under different shear rates ，plasma viscosity ， erythrocyte aggregation index and hematocrit of model group were increased significantly ，while erythrocyte deformation index was significantly decreased ，PT，TT and APTT were significantly shortened （P＜0.01）. Compared with model group ，whole blood viscosity under different shear rates ，plasma viscosity ，erythrocyte aggregation index and hematocrit of aspirin group and W. pigra hang-dried product ，talcum powder-ironed product ，wine bran-processed product high-dose groups were decreased significantly ， while erythrocyte deformation index were significantly increased ，and PT （only W. pigra talcum powder-ironed products high-dose group），APTT（except for W. pigra hang-dried products high-dose group ）and TT were prolonged significantly. The whole blood viscosity of W. pigra hang-dried product medium-dose group under low shear rate ，and those of W. pigra talcum powder-ironed product low-dose ，wine bran-processed product medium-dose groups under low and medium shear rates were decreased significantly. Erythrocyte deformation index of W. pigra talcum powder-ironed product medium-dose group was increased significantly ，while erythrocyte aggregation index was decreased significantly ，and PT ，TT were prolonged significantly. APTT of W. pigra hang-dried product medium-dose group was prolonged significantly. Hematocrit of W. pigra wine bran-processed product low-dose group was decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ： W. pigra hang-dried， talcum powder-ironed and wine bran-processed product can effectively improve hemorheology indexes and prolong blood coagulation time.

Objective:This study is to investigate the job satisfaction of nursing staff in public hospital and ana-lyze the influential factors, thus to provide evidence for improving the performance of human resource management in the hospitals. Methods:A sample of 100 nurses in the affiliated hospital of Weifang Medical University was se-lected to participate in this study with cluster sampling method. Job satisfaction was investigated using self-de-signed questionnaire, and data analysis was conducted with SPSS software. Results:The total score of job satisfac-tion is(92. 52 ± 18. 17). The highest is about their working group with 3. 89 ± 0. 58, while the lowest is about their labor return with(2. 19 ± 0. 89). Authorized strength, education, working years and monthly salary all impact the job satisfaction of nurses. Conclusions:At present, nurses′job satisfaction is at a media level. The managers of public hospitals should build a scientific compensation system and humanized duty system and so on, which can meet the different needs of nursing staff and improve their job satisfaction.

Objective To analyze the clinical characteristics of children tsutsugamushi infection in order to strengthen understanding,reduce the misdiagnosis rate,reduce the complications and improve the level of diagnosis and treatment.Methods To analyze retrospectively the clinical data and treatment outcome of 22 cases of children tsutsugamushi disease,including symptoms,physical signs and laboratory examination.Results All the cases with fever onset,high fever (＞ 39 ℃) was 72.7％ (16/22),all cases were typical eschars and ulcers,while 8 cases (36.4％,8/22)had lymphadenectasis.Merger multiple organ damage in 14 cases (63.6％,14/22),anemia in 11 cases (50.0％,11/22)and liver function abnormal in 4 cases (18.2％,4/22).All of 22 cases were treated with azithromycin.All cases after treatment with azithromycin heal,no recurrence.Conclusions The manifestation of tsutsugamushi disease is diversity,much complications,among which hematology and liver are the most common cases.Physical examination roundly might reduce missed diagnosis and misdiagnosis as greatly as possible.Azithromycin has good therapeutic effects on children tsutsgamushi disease.