Objective To study effect of pentanuclear copper complexes of imino acetic acid two aromatic derivatives on oxidative cutting DNA. Methods Synthesized complexes later, protonation constants and the stability constants of the complexes were determined by potentiometric titration method, the cutting DNA was studied by agarose gelelectrophoresis.Results Under physiological conditions, complexes H4L1, H4L2, between H6L3 and Cu2 +coordination capability was stronger; three carboxylic multi copper complexes in Cu2 + concentration cutting efficiency were higher than the corresponding mononuclear copper complexes, synergistic effect exists between Cu2 + Center; H6L3 complexes of the three DNA cut the highest efficiency, complex H4L1 was followed, complex H4L2 cutting efficiency was the lowest.Conclusion Change the complex chelating ligands has great influence on the properties of complex, which provides a theoretical basis for the design and synthesis of copper nucleases reasonable.

OBJECTIVE: To prepare a ternary supersaturated indomethacin (IND) solid dispersion and investigate its characteristics by screening carrier materials with different functions based on quality by design (QbD) concept. METHODS: Based on dissolution tests, the carriers with solubilization function were selected and the appropriate drug loading ratio of binary solid dispersion was determined. The material as inhibitor of precipitation was chosen using the solvent shift method. The IND ternary supersaturated solid dispersion was prepared by the hot melt extrusion of IND, solubilization material and precipitation inhibitor. The dispersion state of IND was identified by differential scanning calorimetry and powder X-ray diffraction, and its characteristics were explored by the powder wettability test and the stability test. RESULTS: The binary IND solid dispersion with Eudragit EPO could sharply increase IND dissolution rate with a behavior of more than 80%dissolution within 5 min but subsequently followed an obvious concentration decline. Kollidon VA64 had a satisfactory effect of precipitation inhibition for IND supersaturated solution in that the concentration of 0.1% could keep the 50 μg•mL-1 IND solution unchanged within 30 min. The ternary solid dispersion with a mass ratio of 1:2:0.3 (IND:EPO:VA64) could significantly increase the dissolution of IND, eliminating the crystallization and precipitation of the drug in the supersaturated system during the dissolution process and enabling the drug to maintain amorphous form within 3 months. CONCLUSION: Based on the understanding of the functions of different carrier materials and the QbD concept, it could effectively improve the formulation design of solid dispersions. The prepared ternary IND solid dispersions have excellent drug dissolution behavior.

Objective To explore the differences of Th17 population and serum transforming growth factor (TGF)-β1 levels between early-and late-stage primary biliary cirrhosis (PBC) and their roles in pathogenesis.Methods Peripheral Th17 counts were analyzed by flow cytometry.The expression of IL-17A in peripheral blood mononuclear cells and TGF-β1 were measured by real-time quantitative polymerase chain reaction.Serum concentration of TGF-β1 was measured by enzyme-linked immunosorbent assay.Liver biopsies were stained with hematoxylin-eosin to determine the pathological stage.Results were evaluated using KrustalWallis test followed by Mann-Whitney U tests for comparisons of Th17 population between patients with early and late PBC,patients with chronic hepatitis B (CHB) and health controls (HCs).ANOVA followed by LSD t-tests were used for comparing IL-17 mRNA,TGF-β1 mRNA and TGF-β1 serum concentration between groups.The correlations between Mayo risk score and peripheral Th17 of PBC patients,Mayo risk score and serum concentration of TGF-β1 was analyzed by Pearson correlation analysis separately.Results The peripheral Th17 population increased in patients with early PBC (1.03±0.33)％,compared to those with late PBC [(0.48± 0.13％,U=14.0,P＜0.01],CHB [(0.56±0.35)％,U=104.5,P＜0.01],and HCs [(0.36±0.17)％,U=8.0,P＜0.01],while TGF-β1 changed in the opposite direction.Serum concentration of TGF-β1 elevated in late PBC (43.0± 18.7) ng/ml compared with early PBC (29.5±12.2) ng/ml,t=2.85,P=0.006.Conclusion The opposite changes of Th17 population and TGF-β1 level in early and late PBC indicated their different roles in different stages.Th17 may contribute to the autoimmune response in early PBC,participate in the occurrence of autoimmune inflammation,while TGF-β1 to fibrogenesis in late stage.In addition,the possible regulation mechanisms of differentiation of Th17 by TGF-β1 cannot be ignored.

Objective To compare the effects on blood specimen collection from neonatal heel by oblique puncture and vertical puncture methods,to increase the efficiency of puncturing in blood collection.Methods Two hundred eutocia,full-term infants were randomly divided into experimental group (oblique puncture) and control group( vertical puncture) by stochastically ballot,100 samples in each group,and the blood collection rate in two groups was compared.Results In the same condition,both experiment group (87 cases) and control group (56 cases) have higher efficiency in blood collection,but the experiment group is notably easier than control group (x2 =8.76,P＜0.01),however there is no significant difference in pain (t =0.55,P =0.58).Further,we observed the response of neonatal heel and found that both methods will not induce inflammation,but the peidnoma was significantly decreased in experiment group compared to control group ( x2 =9.26,P ＜ 0.01 ).The experiment group was significant high than control group.Conclusions Oblique puncture method is conformance to care program,and can significantly decrease the suffering of newborns,and increase the efficiency.

Objective: To evaluate the clinical efficacy of sinew-regulating bone-setting manipulations plus exercise therapy in treating chronic non-specific low back pain (CNLBP). Methods: A total of 65 CNLBP patients were divided into two groups by the random number table method. Thirty-three cases in the treatment group were intervened by sinew-regulating bone-setting manipulations plus exercise therapy; 32 cases in the control group were intervened by medium-frequency electrotherapy plus exercise therapy. Before and after treatment, visual analog scale (VAS), dynamic and static muscle endurance of low back, median frequency (MF) of surface electromyography (sEMG) and Oswestry disability index (ODI) were used to evaluate the low back function. The therapeutic efficacy was estimated after treatment. Results: The two groups each had 2 dropouts during the study. The total effective rate was 90.3％ in the treatment group versus 66.7％ in the control group, and the between-group difference was statistically significant (P<0.05). After treatment, the VAS score, dynamic and static muscle endurance of low back, MF of sEMG and ODI score all changed significantly in both groups (all P<0.05); all the items in the treatment group were significantly different from those in the control group (all P<0.05). Conclusion: Sinew-regulating bone-setting manipulations plus exercise therapy can effectively release pain in CNLBP patients, increase muscle endurance of the low back and improve the quality of life, and its therapeutic efficacy is more significant than that of medium-frequency electrotherapy plus exercise therapy.

Using a H₂O₂-induced BRL cell senescence model, we investigated the anti-aging effects of drug-containing serums of Erzhi Wan (EZW) and various polar extracts (petroleum ether, ethyl acetate, n-butanol, water, and iridoid glycoside-enriched fractions). Cell viability was detected by MTT assay. Cell senescence was evaluated with β-galactosidase staining assay. Intracellular reactive oxygen species (ROS) were measured by flow cytometry. UFLC-Q-TOF-MS/MS was used to identify chemical components in EZW and the extracts, and molecular docking technology was used to predict the anti-aging components of EZW. Results showed that treatment of cells with 600 μmol·L^-1 H₂O₂ for 72 h markedly induced cell senescence, inhibited cell proliferation and increased intracellular β-galactosidase activity and ROS levels. If cells were pretreated with drug-containing serum of EZW this induction of senescence was decreased. A total of 49 chemical compounds were identified in EZW by liquid chromatography-mass spectrometry, 14 of these were identified by molecular docking as potential active ingredients. Based on these analyses, and the in vitro experiments with polar extracts, we conclude that the anti-aging components of EZW are triterpenes, flavonoids and phenyl alcohols, providing a basis for further elucidation of the active agents and mechanism of the anti-aging effect of EZW.

Objective To study the expression of signal transducer and activator of transcription 3 (STAT3) and Suppressor of cytokine signaling 3(SOCS3) in the middle ear cholesteatoma epithelium ,and the possible roles of STAT3 and SOCS3 in middle ear cholesteatoma .Methods The immunohistochemical assay was used to detect ex-pression of STAT3 and SOCS3 protein in 30 cases of middle ear cholesteatoma epithelial tissue and 20 cases of nor-mal external auditory canal skin tissues as the control group .Results STAT3 immunoreactivity was detected in the nuclei and cytoplasm of epithelial cells .The expression rates of STAT3 in middle ear cholesteatoma epithelial tissue were 76 .7% and higher than in the normal epithelium (25 .0% ) .The differences between the two groups were sta-tistically significant (P<0 .05) .SOCS3 immunoreactivity was detected in the cytoplasm of epithelial cells .The ex-pression rates of SOCS3 in middle ear cholesteatoma epithelial tissue were 33 .3% and lower than in the normal epi-thelium (65 .0% ) .The differences were statistically significant (P<0 .05) .The expression of STAT3 and SOCS3 in the middle ear cholesteatoma had negative correlation (r= - 0 .476 ,P<0 .05) .Conclusion The abnormal ex-pression of STAT3 and SOCS3 in the middle ear cholesteatoma may be involved in hyper proliferation and anti -ap-optosis of cholesteatoma cell ,and play an important role in the formation and development of middle ear cholesteatoma .

Abstract：Objective To investigate the morphological features of the Pneumocystis jirovecii, in order to facilitate early detection and rapid diagnosis of this rare pathogen from a morphology point of view by laboratory technicians. By analyzing the laboratory features and application value of different pathogen detection methods in the diagnosis of Pneumocystis jirovecii pneumonia, we aim to provide the most reliable diagnostic basis for rapid diagnosis of Pneumocystis jirovecii pneumonia.Methods A retrospective analysis was conducted on the test results of bronchoalveolar lavage fluid samples from a comprehensive hospital in Zhangqiu District, Jinan City, Shandong Province, and a hospital in Changde City from April 2022 to October 2022. Five confirmed cases of Pneumocystis jirovecii pneumonia were detected. Its clinical manifestations, laboratory results, and morphological characteristics of pathogens under different stains were analyzed to discuss the advantages and disadvantages of different detection methods. Results Cytological examination of bronchoalveolar lavage fluid found the trophozoites and cysts of Pneumocystis jirovecii by Wright's-Giemsa staining in 4 cases (80%), and the cysts of Pneumocystis jirovecii by Silver hexamine staining in 4 cases (80%), while the metagenomic next-generation sequencing confirmed all the 5 positive results. All 5 patients had different degrees of reduction in the absolute count of peripheral blood lymphocytes, and the serum lactic dehydrogenase and (1-3)-β-D-Glucan were increased. Among the 5 patients in this study, 4 were treated with sulfamethoxazole combined with caspofungin, and 1 was treated with sulfamethoxazole. Three patients were cured and discharged from hospital after treatment, but two died. Conclusions The method of Wright's-Giemsa staining for the cytological examination of bronchoalveolar lavage fluid to find Pneumocystis jirovecii has the unique and irreplaceable advantages as silver staining. Metagenomic next-generation sequencing can further increase the positive detection rate of Pneumocystis jirovecii. The combination of cytological examination of bronchoalveolar lavage fluid with metagenomic nextgeneration sequencing is a powerful diagnostic method for rapid diagnosis of Pneumocystis jirovecii pneumonia, which can diagnose accurately and reduce missed diagnosis.

Radiotracer techniques were employed to characterize 65Zn adsorption and desorption in root-cell-wall of hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end ofa 30 min short time radioisotope loading period, comparable amounts of 65Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more 65Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of 65Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more 65Zn accumulated in desorbed root-cell-wall ofNHE. Zn2+ binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn2+ concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn2+, whereas the desorption characteristics were different, and with most of 65Zn binding on root of HE being available for loading into the xylem, as a result, more 65Zn was translocated to the shoot.

Objective: To observe the therapeutic effect of sinew-regulating bone-setting manipulations for knee osteoarthritis (KOA) model rabbits and its impacts on the chondrocyte apoptosis rate and the levels of interleukin (IL)-1β and nitric oxide (NO). Methods: According to the random number table method, 30 New Zealand white rabbits were divided into a normal group (n=9) and a modeling group (n=21). Rabbits in the modeling group were used to establish KOA models with the modified Hulth method. At the 8th week, three rabbits were sacrificed to verify the model and the remaining 18 rabbits were randomly divided into a model group (n=9) and an intervention group (n=9). Rabbits in the normal group and model group were bred routinely without any intervention. Rabbits in the intervention group were treated with the sinew-regulating bone-setting manipulations, 10 min/time, once every other day for a total of 20 times. The Lequesne MG knee function rating was used to evaluate the behavioral differences of the rabbits in each group. The Pelletier score was used to evaluate the general changes of the rabbits. The Mankin score was used to evaluate the pathology of knee cartilages. The enzyme-linked immunosorbent assay and nitrate reductase methods were used to determine the levels of IL-1β and NO in serum and synovial fluid of each group, respectively. In situ terminal deoxynucleotidyl transferase-mediated nick and labeling method was used to determine the apoptosis of chondrocytes in each group. Results: Compared with the normal group, the scores of Lequesne MG, Pelletier and Mankin, and the levels of IL-1β and NO in the model group were increased (P<0.05), which indirectly indicated the success of the model. Compared with the model group, the scores of Lequesne MG, Pelletier and Mankin, IL-1β and NO levels, and chondrocyte apoptosis rate of the intervention group were decreased, and the differences were statistically significant (P<0.05). Conclusion: The sinew-regulating bone-setting manipulations can reduce the levels of IL-1β, NO, and chondrocyte apoptosis rate, and delay the articular cartilage degeneration, therefore, having a good therapeutic effect on KOA.