The main protease (M^pro) of SARS-CoV-2 is an attractive target in anti-COVID-19 therapy for its high conservation and major role in the virus life cycle. The covalent M^pro inhibitor nirmatrelvir (in combination with ritonavir, a pharmacokinetic enhancer) and the non-covalent inhibitor ensitrelvir have shown efficacy in clinical trials and have been approved for therapeutic use. Effective antiviral drugs are needed to fight the pandemic, while non-covalent M^pro inhibitors could be promising alternatives due to their high selectivity and favorable druggability. Numerous non-covalent M^pro inhibitors with desirable properties have been developed based on available crystal structures of M^pro. In this article, we describe medicinal chemistry strategies applied for the discovery and optimization of non-covalent M^pro inhibitors, followed by a general overview and critical analysis of the available information. Prospective viewpoints and insights into current strategies for the development of non-covalent M^pro inhibitors are also discussed.

Objective To observe the clinical efficacy of joint needling method combined with ultrasound in the treatment of qi stagnation and blood stasis type of patellofemoral pain syndrome(PFPS).Methods Eighty-six patients with qi stagnation and blood stasis type of PFPS were randomly divided into observation group and control group,with 43 cases in each group.The control group was given western medicine conventional treatment combined with functional exercise,and the observation group was given joint needling method combined with ultrasound treatment on the basis of the control group.Both groups were treated for 2 consecutive weeks.After 2 weeks of treatment,the clinical efficacy of the two groups was evaluated,and the changes in the Visual Analogue Scale(VAS)scores of knee pain and the Kujala scale scores of the two groups were observed before and after treatment.The changes in active range of motion(AROM)of the affected knee joint were compared before and after treatment between the two groups.Results(1)After treatment,the VAS scores of the two groups of patients were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the level of VAS scores,and the difference was statistically significant(P＜0.05).(2)After treatment,the Kujala scores of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the level of Kujala scores,and the difference was statistically significant(P＜0.05).(3)After treatment,the AROM of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the level of AROM,and the difference was statistically significant(P＜0.05).(4)The total effective rate was 95.35%(41/43)in the observation group and 81.40%(35/43)in the control group.The efficacy of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion The joint needling method combined with ultrasound can significantly relieve the pain symptoms of patients with PFPS and promote the recovery of knee joint function,and the clinical efficacy is remarkable.

Objective:To analyze the differentially expressed genes in PBMCs of patients with systemic lupus erythematosus (SLE) by bioinformatics methods screening and analyzing the key genes related to ferroptosis, and explore the possible mechanism of ferroptosis involved in the pathogenesis of SLE at the transcription level.Methods:The data sets and samples of healthy people (HC) and SLE patients who met the screening criteria were retrieved from the Gene Expression Omnibus (GEO), a sub-database of the National Center for Biotechnology Information (NCBI). The differentially expressed genes, GO enrichment analysis and KEGG pathway enrichment analysis were analyzed by GEO2R, R language and related software packages. The protein interaction network (PPI) of differential genes was analyzed by STRING, Cytoscape and other tools to explore the key genes and pathways. In addition, real-time quantitative reverse transcription PCR (RT-qPCR) was used to verify the expression of key genes. Mann-Whitney U test was used to compare the expression of key genes in PBMCs between the two groups. Spearman rank correlation analysis was used to explore the relationship between SLE disease activity and the level of key genes. Results:Six data sets were included in this study. A total of 166 genes related to ferroptosis were differentially expressed between SLE and HC groups. The differential genes were specifically expressed in alveolar macrophages, neutrophils, CD49 + cells and CD31 + cells. GO enrichment analysis and KEGG pathway enrichment analysis showed that the differentially expressed genes were mainly involved in multiple signaling pathways closely related to SLE, such as oxidative stress response, infection and TNF signaling pathway. Hub genes screened by different algorithms all suggested RELA as a key gene, and RT-qPCR confirmed that compared with the RELA gene expression level in the HC group [0.75(0.37,1.13)], the expression level in SLE group [2.02 (1.19,4.06)] was increased, the difference was statistically significant ( Z=-3.08, P=0.002), and was positively correlated with the corresponding SLEDAI score of SLE samples ( r=0.52, P=0.019). Conclusion:The ferroptosis of many immune cells, including alveolar macrophages and CD49 + NK cells, is involved in the pathogenesis of SLE. RELA may be involved in the ferroptosis of PBMCs in SLE through the NF-κB pathway.

ObjectiveBased on the quality evaluation experience of "it is better to have a fragrant and strong aroma" summarized by materia medica of past dynasties， the chemical components of Sojae Semen Nigrum（SSN） and Sojae Semen Praeparatum（SSP） were systematically compared and analyzed， and the main fermentation products in different fermentation time were quantitatively analyzed， so as to clarify the transformation law of internal components in the processing process and provide scientific basis for the modern quality control of SSP. MethodUltra performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used for the structural identification of the chemical constituents of SSN and SSP， and with the aid of Progenesis QI v2.3 software， the negative ion mode was employed for principal component analysis（PCA） pattern recognition， and the data were analyzed with the aid of orthogonal partial least squares-discriminant analysis（OPLS-DA） for two-dimensional data to obtain S-plot， and components with |P|>0.1 were selected as the differential constituents. The contents of isoflavonoids in SSP during fermentation was determined by UPLC， and the samples were taken every 8 h in the pre-fermentation period and every 2 d in the post-fermentation period， and the dynamic changes of isoflavonoid contents in different fermentation stages were analyzed. The contents of amino acids and nucleosides in SSP and SSN from different fermentation stages were quantitatively analyzed by phenyl isothiocyanate（PITC） pre-column derivatization and high performance liquid chromatography（HPLC） gradient elution， and the contribution of flavor substances to the "delicious" taste of SSP was discussed by taste intensity value（TAV）. ResultA total of 19 kinds of differential components were screened out， mainly soybean saponins and isoflavones， and their contents decreased significantly or even disappeared after fermentation. In the pre-fermentation process of SSP， glycoside bond hydrolysis mainly occurred， and isoflavone glycosides in SSN were degraded and converted into the corresponding aglycones， the content of flavor substances such as amino acids increased gradually. In the post-fermentation process， protein degradation mainly occurred， after 8 d of post-fermentation， the content of isoflavones was basically stable， while the total content of amino acids increased by 8-40 times on average. Different amino acids form the special flavor of SSP， such as the TAV of glutamate is always ahead of other flavor substances， and sweet substances such as alanine and valine have made relatively great contributions to SSP. ConclusionBased on the law of constituent transformation， combined with the traditional evaluation index of "fragrant and strong"， it is difficult to control the fermentation degree of SSP by the existing standards in the 2020 edition of Chinese Pharmacopoeia. It is suggested that description of the characteristics of SSP be refined and changed to "fragrant， delicious and slightly sweet"， and at the same time， the post-fermentation index compounds such as glutamic acid， alanine and valine should be added as the quality control indicators of SSP， so as to standardize the production process and improve the quality of SSP.

【Objective】 To compare the degree, efficacy and safety of testicular catch-up growth after inguinal and subinguinal surgery for adolescent varicocele. 【Methods】 Clinical data of 121 adolescent varicocele patients with secondary testicular atrophy treated during Sep.2014 and Jun.2021 at Huili People’s Hospital were retrospectively analyzed, including 40 cases treated with the inguinal incision approach, and 81 with the subinguinal approach. The characteristics of intraoperative, efficacy, surgical complications and prognosis were compared between the two groups. 【Results】 The inguinal group had significantly shorter microscope usage time (P=0.006), fewer number of preserved internal spermatic arteries (P<0.001), and significantly larger diameter of preserved internal spermatic arteries than the subinguinal group (P=0.005). The subinguinal group required ligation of more internal spermatic veins, and there were significantly more internal spermatic veins with a diameter of 4 mm or more (P<0.001). No recurrence occurred, and the incidences of scrotal edema and hydrocele were very low (P>0.05). Two years after surgery, the bilateral testicular volume of both groups was significantly larger than that before surgery (P<0.05), and there was no significant difference in the testicles (P>0.05). 【Conclusion】 In the treatment of adolescent varicocele patients with secondary testicular atrophy, the subinguinal approach significantly reduces the difficulty of microsurgical varicocele ligation, and has good efficacy and safety, which is worthy of clinical promotion and application.

Objective:To design an improved lamina hook system and compare its biomechanical properties with traditional lamina hook system in fixation of lumbar spondylolysis.Methods:The thin layer CT data of the lumbosacral vertebrae of 20 healthy young male servicemen who underwent physical examination in the outpatient department of the 940th Hospital of Joint Logistics Support Force of PLA from January 2021 to August 2022 were collected. The age of the subjects was 20-30 years [(25.0±3.0)years]. A 3-dimensional model of the L 5 vertebral body was constructed using the 3-dimensional modeling software. The new improved lamina hook was designed according to the measurements including the thickness of the middle area, the longest longitudinal diameter, the curvature radius of the lower edge, the angle between the upper and lower tail ends, the thickness of the lower edge, and the longest diameter of the lower edge of the bilateral L 5 vertebral plates. One serviceman was selected from the aforementioned group to construct a linear finite element model of segments L 4-S using the 3-dimensional virtual software (normal model, model A), based on which, the L 5 bilateral spondylolysis model (model B), improved lamina hook model (model C) and traditional lamina hook models (model D) were designed. By constraining both sides of the sacrum and applying a longitudinal load of 400 N on the L 4 vertebral body, the upper 1/3 gravity of the body was simulated, and with a bending moment of 10 N·m along the X, Y, and Z directions, motions of forward flexion, backward extension, lateral bending, rotation, etc were simulated. The range of motion of segment L 4/5 and L 5/S 1 of model A was evaluated and compared with the findings of the previous researches to verify its effectiveness. The overall range of motion of models A, B, C, and D, the range of motion of segment L 4/5 and L 5/S 1, the maximum overall displacement, the maximum displacement and stress of the isthmus, the stress distribution and maximum stress of internal fixation of models C and D, and the stress distribution and maximum stress of the vertebral body of models C and D were compared. Results:(1) During forward flexion, backward extension, lateral bending and rotation, the range of motion of model A was 5.01°, 4.03°, 3.91° and 1.42° in segment L 4/5, and was 4.62°, 2.51°, 2.40° and 1.23° in segment L 5/S 1. (2) The overall range of motion, range of motion of segment L 4/5 and L 5/S 1 and maximum overall displacement of models A, C, and D were similar in axial compression, forward flexion, backward extension, left bending, and left rotation, while those of model B were significantly increased. (3) There was no significant difference in the maximum displacement of the isthmus of models A, C, and D under different motion modes, while the maximum displacement of model B in the isthmus was significantly larger than that of models A, C, and D, especially during rotation, increased by 295%, 277%, and 276% respectively. The maximum stress of the isthmus of model C was 0.938 MPa, 1.698 MPa, 0.410 MPa, 2.775 MPa, and 1.554 MPa respectively. The maximum stress in the isthmus of model D was 0.590 MPa, 1.297 MPa, 0.520 MPa, 3.088 MPa, and 2.072 MPa respectively. The maximum stress of the isthmus of models C and D was similar during axial compression and forward flexion, while the stress of the isthmus of model C was smaller than that of model D during backward extension, lateral bending, and rotation, decreased by 21.1%, 10.2%, and 25.0% respectively compared with model D. (4) The maximum stress of internal fixation in models C and D during forward flexion, backward extension, left bending, and left rotation was 135.220 MPa, 130.180 MPa, 200.940 MPa and 306.340 MPa respectively, and was 131.840 MPa, 112.280 MPa, 349.980 MPa and 370.140 MPa respectively. The maximum stress of internal fixation in the two models of internal fixation during forward flexion and backward extension was similar, while it was decreased by 42.6% and 17.2% in model C during left bending and left rotation, compared with model D. (5) The maximum stress of the vertebral body during forward flexion, backward extension, left bending, and left rotation was 79.787 MPa, 36.857 MPa, 37.943 MPa and 96.965 MPa respectively in model C, but was 80.104 MPa, 64.236 MPa, 196.010 MPa and 193.020 MPa respectively in model D. The maximum stress of models C and D was all distributed in the contact area with the internal fixation, and especially during backward extension, left bending, and left rotation, when it was reduced by 42.6%, 80.6%, and 49.8% of model C respectively, compared with that of model D. Conclusions:The improved laminar hook is more consistent with the Chinese anatomized structure of the lamina. Compared with the traditional lamina hook system, the improved lamina hook system can effectively reduce the displacement in all directions and range of motion of lumbar spondylolysis, therefor can significantly reduce the stress of internal fixation and vertebral body and has better biomechanical performance.

Objective:To summarize the imaging characteristics of primary intestinal lymphoma (PIL).Methods:As a cross-sectional study, a total of 103 consecutive patients diagnosed as PIL by pathology were retrospectively enrolled at Affiliated Shandong Provincial Hospital of Shandong First Medical University between January 2013 to December 2023. Ninety-four patients underwent CT plain and enhanced scans, and the remaining 9 patients only underwent MRI examination. Imaging features were evaluated, including the location and shape of the tumor, the shape of the intestinal wall and lumen, abdominal lymph nodes, the density or signal intensity of the tumor, and the degree and mode of enhancement after the enhancement scan.Results:The most frequent location of PIL was the ileum (43.7%, 45/103), followed by the ascending colon (20.4%, 21/103). Ninety-one patients had a single lesion (88.3%), while the other 12 patients had multiple lesions (11.7%). Diffuse large B-cell lymphoma was the most common histopathological type of the PIL (65.0%, 67/103), followed by mucosa-associated lymphoid tissue lymphoma (9.7%, 10/103). Generally, PIL classically presented intestinal wall thickening (81.6%, 84/103) and exophytic mass (28.2%, 29/103). Aneurysmal dilatation of the lumen appeared in 19 patients. Nineteen tumors (18.4%) grew out of the lumen, and perforation was shown in 3 patients. Bulky mesenteric or retroperitoneal adenopathy occurred in 28 patients (27.2%). Seventy-two tumors (69.9%) showed homogeneous enhancement, and 75 (72.8%) showed mild-to-moderate enhancement. In DWI images, all 9 tumors showed significant diffusion limitation.Conclusions:The most frequent location of PIL was the ileum. The main imaging characteristics were segmental intestinal wall thickening, which showed mild-to-moderate homogeneous enhancement and restricted diffusion.

Objective:To explore the value of whole exome sequencing for the inferential analysis of recessive genetic disease carrier status for couples with a child died of Primary immunodeficiency (PID).Methods:Clinical data was collected from four couples with a childbearing history of PID who had sought genetic counseling and undergone genetic testing at Henan Provincial People′s Hospital from February 2017 to December 2021. Whole exome sequencing (WES) was performed on both partners of each couple, and candidate variants were validated by Sanger sequencing and fluorescent quantitative PCR. Prenatal diagnosis was conducted on fetuses of these couples after confirming the variants.Results:A total of six variants were detected in four genes including IL2RG, BTK, CYBB, and DUOX2. Among these, the c.1265G>A and c.3329G>A variants of the DUOX2 gene and the c. 676C>T variant of the IL2RG gene were previously known as pathogenic variants. On the other hand, the Exon5_8del variant of the IL2RG gene, the c. 184_185delAC variant of the BTK gene, and the c. 472A>T variant of the CYBB gene were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics, the IL2RG: Exon5_8del, BTK: c. 184_185delAC and CYBB: c. 472A>T variants were classified as likely pathogenic (PVS1+ PM2_Supporting+ PP4).Prenatal diagnosis was conducted for three couples during their subsequent pregnancies, and the results revealed that the fetuses had the wild-type genotypes at the c. 184_185 position of the BTK gene, the c. 472 position of the CYBB gene, and the c. 676 position of the IL2RG gene. Follow-up examinations one year after birth has found no abnormality in the infants. Conclusion:WES is an important tool to infer and analyze the carryier status for couples who had given births to children died of PID and improve the positive detection rate.

This study was aimed at investigating the effect of lymphocyte activation gene-3(LAG3)on liver B lymphocyte subsets and their functions in WT and LAG3-KO mice infected with Echinococcus multilocularis(E.multilocularis).In a mouse model of E.multilocularis infection,the expression and localization of CD19 and α-SMA in liver were detected by immu nohistochemistry.CD80,CD86 and MHC-Ⅱ molecules expressed on B cells and their subsets in mice liver were detected by flow cytometry.After 12 weeks of infection,the area and percentage of CD19 in LAG3-KO group was slightly higher than that in WT group,but the difference was not statistically(t=-1.241、-1.237,P＞0.05).The area and percentage of a-SMA in LAG3-KO group was higher than that in WT group(t=-3.224、-3.227,P＜0.05).The proportion of CD80 and MHC-Ⅱ molecules expressed on liver B cells in LAG3-KO group was up-regulated(t=-2.379,-3.321,P＜0.05).The percentage of liver B2 cells in LAG3-KO group was higher than that in WT group(t=-2.695,P＜0.05).The expression of CD80 on Blb cells in LAG3-KO group was significantly up-regulated(t=-5.315,P＜0.001).The proportion of CD80 of B2 cells in LAG3-KO group was lower than that in WT group(t=2.806,P＜0.05).The expression of MHC-Ⅱ molecule in B2 cells in LAG3-KO group was up-regulated(t=-4.227,P＜0.01).It is suggested that LAG3 molecules affected the B cell subsets and func-tion of mouse liver in the middle stage of E.multilocularis infection,especially B2 lymphocytes.LAG3 molecule exerted an in-hibitory effect on the activation of B cells and the expression of MHC-class Ⅱ molecules,suggesting that it may be involved in B cell exhaustion caused by E.multilocularis.

BACKGROUND: Craniomaxillofacial tissue defects are clinical defects involving craniomaxillofacial and oral soft and hard tissues.They are characterized by defect-shaped irregularities, bacterial and inflammatory environments, and the need for functional recovery.Conventional clinical treatments are currently unable to achieve regeneration of high-quality oral craniomaxillofacial tissue. As a natural biomaterial, silk fibroin (SF) has been widely studied in biomedicine and has broad prospects for use in tissue regeneration.Hydrogels made of SF showed excellent water retention, biocompatibility, safety and the ability to combine with other materials. METHODS: To gain an in-depth understanding of the current development of SF, this article reviews the structure, preparation and application prospects in oral and craniomaxillofacial tissue regenerative medicine. It first briefly introduces the structure of SF and then summarizes the principles, advantages and disadvantages of the different cross-linking methods (physical cross-linking, chemical cross-linking and double network structure) of SF. Finally, the existing research on the use of SF in tissue engineering and the prospects of using SF with different cross-linking methods in oral and craniomaxillofacial tissue regeneration are also discussed. CONCLUSIONS This review is intended to show the advantages of SF hydrogels in tissue engineering and provides theoretical support for establishing novel and viable silk protein hydrogels for regeneration.