Senile osteoporosis (SOP) is a systemic bone disease characterized by increased susceptibility to fractures. The pathogenesis of SOP is complex and not well understood. Currently, the rapid aging model mouse, senescence accelerated mouse prone 6 (SAMP6), is an ideal model for studying the mechanisms of SOP development and exploring its prevention and treatment. This model exhibits characteristics including increased bone fragility, degradation of bone microstructure, loss of bone matrix, and abnormal metabolism and dysfunction of bone cells, faithfully replicating the process of SOP occurrence and progression at both macroscopic and microscopic levels.

Objective To explore influencing factors of functional recovery after ankle fracture of type Ⅱ degree and above supination-external rotation.Methods Clinical data of 120 patients with ankle fractures of type Ⅱ degree and above supination-external rotation admitted from February 2019 to April 2021 were retrospectively analyzed.According to American Orthopaedic Foot and Ankle Society(AOFAS),patients were divided into excellent group with 73 patients(90 to 100 points),good group with 35 patients(75 to 89 points),and fair group with 12 patients(＜50 points).The differences of ankle active range of motion(ROM)and AOFAS score were compared among three groups at the latest follow-up.Multivariate Logistic regression analy-sis was performed to analyze the factors related to functional recovery after ankle fracture of supination-external rotation.Results There were significant differences in postoperative ROM(dorsoextension,plantar flexion,varus and valgus)and com-plications between excellent group and good and acceptable group(P＜0.05).Univariate analysis showed there were differences in age above 50 years old,Ⅳ degree of supination-external rotation fracture,lower tibiofibular ligament injury,posterior ankle fracture,no drainage tube placed,infection,antibiotic use time above 7 days(P＜0.05).Multivariate Logistic regression analy-sis showed age above 50 years old[OR=2.829,95％CI(1.049,7.628),P=0.040],Ⅳ degree fracture of supination-external ro-tation[OR=6.13,95％CI(1.153,32.593),P=0.033],lower tibiofibular ligament injury[OR=10.785,95％CI(3.338,34.894),P=0.000],and posterior ankle fracture[OR=6.349,95％CI(1.869,21.560),P=0.003]were independent risk factors for func-tional recovery after ankle fracture of supination-external rotation(P＜0.05).Conclusion The postoperative excellent outcome of ankle fracture was good,and the recovery of joint motion was better.The older age of patient,Ⅳ degree of supination-exter-nal rotation fracture,the lower tibiofibular ligament injury,and posterior ankle fracture are all adverse factors affecting func-tional recovery after supination-external rotation ankle fracture.In clinical,effective measures should be taken to deal with these influencing factors,and strive to improve the functional recovery after the operation of this type of fracture and reduce the occurrence of related complications.

Celastrol and wilforlide A are the main active triterpenoids of the traditional Chinese medicine Lei Gong Teng, which have anti-tumour, anti-inflammatory and immunosuppressive activities, and are the material basis for the clinical efficacy of Lei Gong Teng-related Chinese medicinal preparations. By analysing the biosynthetic pathway of active ingredients, optimizing genetic elements and utilizing "cell factory" to produce triterpenoids heterologously will be an effective way to obtain from Tripterygium wilfordii in the future in a "low-cost and high-efficient" manner. CYP712Ks are the first cytochrome P450s involved in the skeleton modification of friedelane-type and oleanane-type triterpenoids in T. wilfordii, and they can catalyse the generation of polpunonic acid from friedelin and 3-epi-katonic acid from β-amyrin by carboxylation at C-29 position. In this study, four multifunctional TwCYP712K1/2/3/5 were used to clarify the catalytic function and substrate selection preference using in vivo functional characterization in Saccharomyces cerevisiae. The spatial structure of the protein-substrate binding was clarified through homology modeling and molecular docking, and then the differential amino acids in the active pocket of the protein were mutated to clarify the crucial amino acids determining the catalytic function and the selection of substrate structure. A total of 63 mutant elements were constructed, and the amino acid sites affecting the carboxylation function of TwCYP712Ks were analyzed. In particular, the key amino acids affecting the substrate selectivity of TwCYP712K2 towards oleanane-type and friedelane-type triterpenoids were revealed and the TwCYP712K2^F127I and TwCYP712K2^A227T mutants would result in a reversal of the product ratio. In conclusion, four proteins of TwCYP712Ks were semi-rationally designed by homologous protein alignment and mutual mutation to elucidate multiple amino acid sites determining the catalytic function of the proteins, and a series of activity-enhancing or altering mutants were obtained, which provide abundant catalytic elements for the biosynthesis of active triterpenoids from T. wilfordii, and the mechanism of carboxylation in the C-29 position was initially elucidated.

The MADS-box gene family is a very important transcriptional regulator gene, which plays a role in the whole growth and development process of plants. The APETALA1 (AP1) gene is considered to play an important regulatory role in the transformation of plant flowering, but also to control the characteristic development of floral organs. Lonicera macranthoides is used as medicine with dry buds and early flowers. Therefore, studying the potential mechanism of AP1 gene in regulating flower organ development can provide a basis for improving its medicinal value by molecular means. To explore the potential mechanism of the AP1 gene in the regulation of floral organ development in L. macranthoides, the full-length cDNA of the AP1 was cloned by reverse transcription PCR (RT-PCR) and named LmMADS4. The results show that the CDS of the LmMADS4 gene is 729 bp and encodes 242 amino acids, and the LmMADS4 protein contains no signal peptide and no transmembrane structure, which is an unstable hydrophilic protein. Through homologous sequence alignment and phylogenetic analysis, LmMADS4 and L. japonica MADS27 protein cluster into one class and are closely related. Finally, the expression pattern and protein interaction pattern of LmMADS4 were analyzed by real-time reverse transcription-PCR (qRT-PCR) and yeast two-hybrid technology. The qRT-PCR showed that LmMADS4 gene was differentially expressed in the stems, leaves and flower bud at different developmental stages, including bud type variety Longhua and common variety Baiyun; and LmMADS4 gene was highly expressed in the flower buds, and with the development of flower buds, LmMADS4 gene was continuously up-regulated in the flower bud variety Longhua, however, the expression level of LmMADS4 in the Baiyun terminal flower bud was lower than that in the late flower bud, but the difference was not significant. The yeast two-hybrid results showed that the bait vector pGBKT7-LmMADS4 was not toxic to yeast strains and had no self-activating activity. LmMADS4 protein interacted with LmSVP1, LmSVP3 and LmSOC1s proteins. This study can provide a theoretical basis for exploring the mechanism of long flower bud stage and corolla non-unfolding at the molecular level and variety improvement of L. macranthoides.

Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

Objective To explore the clinical value of anti-Müllerian hormone(AMH)to optimize endocrine therapy for peri-menopausal early breast cancer.Methods Two hundred and four patients of pre-menopausal breast cancer aged 45-55 years old between 2020 and 2023 were enrolled,and AMH≤0.1 ng/mL was considered as cut-off value for menopause.Switching from selective estrogen receptor modulator(SERM)to aromatase inhibitor aromatase inhibitor(AI)and initial endocrine therapy regimens were based on AMH,follicle-stimulating hormone(FSH)and estradiol(E2).Results Pre-chemotherapy AMH level was significantly negatively correlated with FSH level(P<0.001).Among 100 cases who were amenorrhea for one year during SERM treatment,42 cases did not have AMH testing.Fourteen out of the 42 cases switched to AI within one year,and ovarian function recovery(OFR)occurred in 2 cases after AI switching.Fifteen cases with AMH>0.1 ng/mL did not switch to AI within one year.Forty among 43 cases with AMH≤0.1 ng/mL switched to AI,after a significantly shorter median SERM treatment duration(3.15 months vs.8.14 months,P<0.001)and a significantly lower OFR rate(0 vs.12.5%,P=0.023)compared with those who did not test AMH but switched to AI.AMH≤0.1 ng/mL was an independent risk factor of transition to menopause shortly in peri-menopausal patients(OR=35.857,P<0.001).Among 104 cases with AMH tested before adjuvant chemotherapy,69 cases had AMH>0.1 ng/mL.Thirty-one out of the 69 cases were treated with ovarian function suppression(OFS)initially and 38 with SERM initially.Thirty-five cases with AMH≤0.1 ng/mL were all treated with SERM initially,with a higher rate of switching to AI(71.4%vs.23.7%,P<0.001)and a shorter SERM treatment duration(6.52 months vs.13.56 months,P=0.016)compared with the 38 cases(AMH>0.1 ng/mL)treated initially with SERM.After a median 30-month follow-up,no recurrence was observed in these thirty-five cases treated with SERM initially and AMH≤0.1 ng/mL,just like in OFS group.And they had a tendency of improved survival outcome compared with those treated with SERM initially and AMH>0.1 ng/mL(Log Rank P=0.076).Conclusion AMH could evaluate and predict menopause accurately,resulting in optimizing endocrine therapy for peri-menopausal patients effectively and safely.

Hereditary breast cancer refers to breast cancer with a genetic susceptibility gene.PTEN germline mutations are rare in breast cancer,but patients with PTEN mutations have a high risk of breast cancer.In 2021,A young patient with bilateral breast cancer was admitted to the Obstetrics and Gynecology Hospital,Fudan University.Due to bilateral multiple breast lumps,she underwent Vacuum-Assisted Breast Biopsy,which was pathologically confirmed as right ductal carcinoma in situ,left breast invasive carcinoma.The patient had multiple neoplasms in bilateral axillary region skin,neck skin and bilateral inguinal regiona skin,and the second-generation sequencing results of peripheral blood genes showed PTEN gene mutation.Combined with family history,the patient was diagnosed with Cowden syndrome(CS).Such patients should be paid attention to cancer risk management and family management,so as to attain early diagnosis and treatment.

Objective To investigate the effect of long non-coding RNA(lncRNA)nuclear paraspeckle assembly transcript 1(NEAT1)in regulating osteogenic differentiation of human bone marrow-derived mesenchymal stem cells(hBMSCs)through regulating cell pyroptosis.Methods hBMSCs were cultured for 7 days to induce osteogenic differentiation,and then divided into the control group(common culture),the osteogenic differentiation group(osteogenic differentiation induction),the pcD-NEAT1 group(transfected with NEAT1 overexpression plasmid),the pcD-null group(transfected with negative control of NEAT1 overexpression plasmid),the osteogenic differentiation+CML group[treated with osteogenic differentiation induction and NOD-like receptor family protein 3(NLRP3)inflammasome activator of carboxy methyl lysine(CML)],and the osteogenic differentiation+CML+pcD-NEAT1 group(treated with osteogenic differentiation and pcD-NEAT1 as well as CML).The cell mineralization was detected using alizarin red staining,the alkaline phosphatase(ALP)activity was determined by ALP activity assay,the cell survival rate was determined by CCK-8,and the morphological change was observed under a high-power microscope.The cell apoptosis was determined using the TUNEL assay.The expression of NEAT1 was detected using qRT-PCR.The protein expression of IL-1β,IL-18,NLRP3,cleaved-caspase 1(cleaved-CASP1),gasdermin D,Runt-related transcription factor 2(RUNX2),ALP,and osteopontin(OPN)were detected using Western blot.Results Compared with the control group,the degree of cell mineralization in the osteogenic differentiation group was increased,the ALP activity was elevated(P<0.05),and the expression of NEAT1 and the protein expression of RUNX2,ALP,and OPN were upregulated(P<0.05).Compared with the pcD-null group,the degree of cell mineralization in the pcD-NEAT1 group was increased,the ALP activity was elevated(P<0.05),and the expression of NEAT1 and the protein expression of RUNX2,ALP,and OPN were upregulated(P<0.05).Compared with the osteogenic differentiation group,the degree of cell mineralization in the osteogenic differentiation+CML group was reduced,the ALP activity was decreased(P<0.05),the cell survival rate was reduced(P<0.05),the cell apoptosis was increased(P<0.05),cell membrane rupture occurred,cells showed enlarged deformation,the expression of NLRP3 and the protein expression of IL-1β,IL-18,cleaved-CASP1,and gasdermin D were significantly upregulated(P<0.05),while the protein expression of RUNX2,ALP and OPN were significantly downregulated(P<0.05).Compared with the osteogenic differentiation+CML group,the degree of cell mineralization in the osteogenic differentiation+CML+pcD-NEAT1 group was increased,the ALP activity was increased(P<0.05),the protein expression of RUNX2,ALP and OPN were significantly upregulated(P<0.05),the cell survival rate was increased(P<0.05),the cell apoptosis rate was decreased(P<0.05),the cell membrane was intact with the normal cell shape,the expression of NLRP3 and the protein expression of IL-1β,IL-18,cleaved-CASP1 and gasdermin D were significantly downregulated(P<0.05).Conclusion The overexpression of NEAT1 promotes the osteogenic differentiation of hBMSCs by inhibiting NLRP3 inflammasome-mediated cell pyroptosis.

Objective:To understand the effect of collagen peptides on the function of mouse lymphocytes under simulated microgravity.Methods:The splenocytes of mice were isolated,and the rotary cell culture system was used to simulate the microgravity.The T lymphocytes were stimulated with mitotic agents,concanavalin A(ConA),and the cells were treated with different concentrations of collagen peptides.The proliferation of lymphocytes and the levels of cytokines in the supernatant were detected.Results:Simulated microgravity could inhibit the proliferation of spleen T lymphocytes and decrease the level of cytokines in the supernatant.Collagen peptides could promote the lymphocyte proliferation and cytokine production in cells cultured under simulated microgravity.Conclusion:Collagen peptides may attenuate the inhibitory effect of simulated microgravity on T lymphocytes by regulating the cell proliferation and the secretion of cytokines.

Surgical robot is the national strategic diagnostic and therapeutic equipment research focus,get a number of scientific research institutes,colleges and universities and enterprises pay extensive attention to the design and development of a variety of surgical robots,and registration declaration.This article explores the critical technologies of surgical robots and key areas for optimizing their performance,including kinematic positioning errors,pose errors,feedback model errors,image recognition positioning errors,path planning,and safety aspects.The findings not only provide a scientific basis for future standardization research on surgical robots but also offer significant theoretical and practical references for the research,manufacturing,and registration processes in the medical robotics industry.