OBJECTIVE To establish a determination method for the related substances of LPM3480392, a novel Gi protein-biased opioid receptor(MOR) agonist. METHODS The separation was carried out with Waters Symmetry Shield RP18 (150 mm×4.6 mm, 3.5 μm) by gradient elution method, using a mixture of 0.002 5 mol·L–1 sodium 1-octanesulfonate monohydrate in 0.01 mol·L–1 potassium dihydrogen phosphate-water solution(containing 0.1% triethylamine, adjusted pH to 2.50 with phosphate acid) and acetonitrile as the mobile phase at a flow rate of 1.0 mL·min–1 and the UV detection wavelength was set at 210 nm. RESULTS The chromatographic peaks of LPM3480392 and impurity A, impurity B, impurity C, impurity E and impurity F could be completely separated, the linear relationship of LPM3480392 was good in 0.064 9−5.191 2 μg·mL–1, while impurity A, impurity B, impurity C, impurity E and impurity F showed good linear relationship within 0.066 6−7.610 4 μg·mL–1, 0.166 0−3.794 0 μg·mL–1, 0.209 2−4.463 2 μg·mL–1, 0.167 9−7.672 6 μg·mL–1 and 0.016 4−7.505 7 μg·mL–1, respectively. The recovery rate was within 93.0%−103.2%. CONCLUSION The method is suitable for the determination of related substances in LPM3480392, and can provide valuable reference for the follow-up research and development of LPM3480392.

Cell mechanics is essential to cell development and function,and its dynamics evolution reflects the physiological state of cells.Here,we investigate the dynamical mechanical properties of single cells under various drug conditions,and present two mathematical approaches to quantitatively character-izing the cell physiological state.It is demonstrated that the cellular mechanical properties upon the drug action increase over time and tend to saturate,and can be mathematically characterized by a linear time-invariant dynamical model.It is shown that the transition matrices of dynamical cell systems signifi-cantly improve the classification accuracies of the cells under different drug actions.Furthermore,it is revealed that there exists a positive linear correlation between the cytoskeleton density and the cellular mechanical properties,and the physiological state of a cell in terms of its cytoskeleton density can be predicted from its mechanical properties by a linear regression model.This study builds a relationship between the cellular mechanical properties and the cellular physiological state,adding information for evaluating drug efficacy.

Objective:To analyze the clinical characteristics of monkeypox patients.Methods:The clinical data and laboratory findings of 4 patients with monkeypox patients diagnosed at Yiwu Central Hospital in July 2023 were analyzed. Herpes fluid and skin tissue samples were collected, the viruses were isolation and cultured in African green monkey kidney cells (Vero) and identified with whole gene sequencing.Results:All four patients were male, aged 24-35 years. All patients had male-to-male behavior within 21 days before onset of the disease. Among them, one patient has AIDS and one patient has syphilis. Four patients presented with perineal skin lesions with itching, and 3 patients were found to have enlarged lymph nodes upon admission. Laboratory testing: lymphocyte abnormality (4.57×10 9/L) in 1 case; increased procalcitonin (0.25 ng/mL) in 1 case; elevated IL-10 levels ( 7.11 ng/L and 9.42 ng/L) in 2 cases; increased IL-6 (66 ng/L) and IL-4 (3.24 ng/L) in 1 case, respectively. One case had abnormal myocardial zymogram with a elevated lactate dehydrogenase level of 313 U/L. The monkeypox virus was isolated from lesion tissue and herpes fluid, and the whole gene sequencing identified it as the B. 1.3 subtype of the IIb evolutionary branch, exhibiting typical pathological effects on Vero cells. Conclusion:The clinical manifestations of the 4 monkeypox patients confirmed in Zhejiang province are mild, patients had a definitive history of male-to-male sexual behavior and the virus strains belong to the B. 1.3 lineage of the IIb evolutionary branch.

Familial acne inversa (AI) is an autoinflammatory disorder that affects hair follicles and is caused by loss-of-function mutations in γ-secretase component genes. We and other researchers showed that nicastrin (NCSTN) is the most frequently mutated gene in familial AI. In this study, we generated a keratin 5-Cre-driven epidermis-specific Ncstn conditional knockout mutant in mice. We determined that this mutant recapitulated the major phenotypes of AI, including hyperkeratosis of hair follicles and inflammation. In Ncstn;K5-Cre mice, the IL-36a expression level markedly increased starting from postnatal day 0 (P0), and this increase occurred much earlier than those of TNF-α, IL-23A, IL-1β, and TLR4. RNA-Seq analysis indicated that Sprr2d, a member of the small proline-rich protein 2 family, in the skin tissues of the Ncstn;K5-Cre mice was also upregulated on P0. Quantitative reverse-transcription polymerase chain reaction showed that other Sprr2 genes had a similar expression pattern. Our findings suggested that IL-36a might be a key inflammatory cytokine in the pathophysiology of AI and involved in the malfunction of the skin barrier in the pathogenesis of AI.

The contractile force of hepatic stellate cells plays a very important role in liver damage, hepatitis and fibrosis. In this paper, a method based on polydimethylsiloxane (PDMS) thin micropillar arrays is proposed to measure the contractile force of human hepatic stellate cell line LX-2, which enables dynamic measurement of the subcellular distribution of force magnitude and direction. First, thin micropillar arrays on glass bottom dish were fabricated using two-step casting process in order to meet the working distance requirement of 100× objective lens. After hydrophilic treatment and protein imprint, cells were seeded on the micropillar arrays. LX-2 cells, which were quiesced by growth in serum-free medium, were activated by adding fetal bovine serum (FBS). The deflections of the micropillars were achieved by image processing technique, and then the contractile force of cells exerted on the micropillars was calculated according to mechanical simulation results, and was analyzed under both quiescent and activated conditions. The experimental results show that the average traction force of quiescent cells is about 20 nN, while the contractile force of activated cells increased to 110 nN upon adding FBS. This method can quantify the contractile force of LX-2 cell on subcellular scale in both quiescent and activated states, which may benefit pathology study and drug screen for chronic liver diseases resulted from liver fibrosis.
Cell Line ; Hepatic Stellate Cells ; cytology ; Humans ; Image Processing, Computer-Assisted ; Mechanical Phenomena

The traditional method of multi-parameter flow data clustering in flow cytometry is to mainly use professional software to manually set the door and circle out the target cells for analysis. The analysis process is complex and professional. Based on this, a clustering algorithm, which is based on -distributed stochastic neighbor embedding ( -SNE) algorithm for multi-parameter stream data, is proposed in the paper. In this algorithm, the Euclidean distance of sample data in high dimensional space is transformed into conditional probability to represent similarity, and the data is reduced to low dimensional space. In this paper, the stained human peripheral blood cells were treated by flow cytometry, and the processed data were derived as experimental sample data. The -SNE algorithm is compared with the kernel principal component analysis (KPCA) dimensionality reduction algorithm, and the main component data obtained by the dimensionality reduction are classified using -means algorithm. The results show that the -SNE algorithm has a good clustering effect on the cell population with asymmetric and trailing distribution, and the clustering accuracy can reach 92.55%, which may be helpful for automatic analysis of multi-color multi-parameter flow data.

By analyzing the effect of 5A group-managementpattern on the community -based diabetes management , the article aimed to establish a new model of community -based diabetes management .Method It de-signed a community-based intervention program , and discussed the intervention effect by comparing the indicators before and after the intervention .Result After 12-months intervention , intervention group patients are better than the contrast group in aspects of mental health , the control of HBAIC and blood pressure , and use of medical resources . Conclusion The 5A group-managementpattern may help the patients in improving mental health controlling blood pressure and HBAIC , and improving medical resources utilization .

Objective To evaluated the application value of two kinds of mass spectrometer(MS)and Vitek MS system in the i-dentification of routinely isolated bacteria in clinic.Methods 149 strains of common bacteria(including 14 genera and 30 species)i-solated from blood,urine,cerebral spinal fluid,secretion and sputum samples in our hospital from March 2012 to January 2013 were collected and simultaneously identified by 2 kinds of matrix-assisted laser desorption ionization-time of flight mass spectrometer (MALD-TOF-MS).The identification results were compared with those identified by the conventional biochemical identification (Vitek2 compact).The strains with the inconsistent results identified by 3 kinds of method were confirmed by 16S rDNA gene se-quencing.Results Among 149 common bacteria,the correct identification rates of genus and species by the Bruker Biotyper MS were 98% and 96% respectively and which by the Vitek MS system were 97% and 95% respectively.There were no misidentified bacterial strains by these two kinds of MS.Conclusion No statistical difference in the identification results was observed between these two kinds of MS system(P >0.05).Both exhibit excellent identification level and are suitable for the routine laboratory iden-tification of clinical microorganism.

Objective To construct the recombinant plasmid of coxsackievirus group A type 16 (CA16)VP1,express in E.coli and identify expressed product. Methods The CA16 VP1 gene was amplified by PCR,The constructed recombi-nant plasmid pET21b-CA16 VP1 was transformed into E.coli BL21(DE3) for expession and purification,expressed and purified product was identified by SDS-PAGE,Indirect ELISA tested Antigenicity and validity of recombinant proteins CA16 VP1. Results The coxsackievirus group A type 16 VP1 protein was expressed in peokaryotic cells and purified, which showed specific reaction with mice antiserum against CA16 virus,high reactivity with part serums of the elderly population in Taiyuan city. Conclusion CA16 VP1 protein was successfully expressed, had good Antigenicity and va-lidity,which will lay a foundation for study on the CA16 diagnostic kits.

Objective To discuss the application of evidence-based nursing in home rehabilitation of elderly patients with breast cancer .Methods Eighty-six elderly patients with breast cancer were randomly divided into the control group and the observation group .The control group received conventional nursing plan after discharge , while the observation group conducted comprehensive nursing evaluation , systematic analysis , nursing intervention methods and implementation according to evidence-based nursing . Changes in living environment of home rehabilitation , social mentality , mentality and health were evaluated by Omaha system . Results The score of recognition, behavior and current situation was respectively ( 7.987 ±3.520 ), (7.341 ±2.895) and (7.756 ±2.903) in the observation group after intervention , which were higher than (4.456 ±4.123),(4.780 ±3.214) and(4.653 ±3.424) in the control group, and the differences were statistically significant (t=4.198,4.495,4.179,respectively;P<0.05).The score of clothing, eating, living, transportation and personal hygiene was respectively (88.462 ±3.132),(96.875 ±1.990),(92.547 ± 2.424),(86.342 ±3.452),(96.867 ±3.094)in the observation group, which were higher than (68.764 ± 3.098),(70.624 ±4.252),(66.245 ±2.870),(73.085 ±2.094),(68.326 ±3.021) in the control group, and the differences were statistically significant ( t =5.569,5.412,5.478,5.398,5.402, respectively;P <0.05).Conclusions Evidence-based nursing is effective for home rehabilitation of elderly patients with breast cancer.