The genome tagging project (GTP) plays a pivotal role in addressing a critical gap in the understanding of protein functions. Within this framework, we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411 (HA-tagged Ssp411) mouse model. This model is instrumental in probing the expression and function of Ssp411. Our research revealed that Ssp411 is expressed in the round spermatids, elongating spermatids, elongated spermatids, and epididymal spermatozoa. The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis. Nevertheless, rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions. Ssp411 is not detectable in metaphase II (MII) oocytes, zygotes, or 2-cell stage embryos, highlighting its intricate role in early embryonic development. These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP, fostering groundbreaking advancements in the fields of spermiogenesis and reproductive biology.
Animals ; Female ; Humans ; Male ; Mice ; Spermatids/metabolism* ; Spermatogenesis/physiology* ; Spermatozoa/metabolism* ; Thioredoxins/genetics*

Objective To conduct a clinical comprehensive evaluation of Lu Dangshen Oral Liquid from 6 dimensions,including safety,effectiveness,economic value,innovativeness,suitability and accessibility.Methods The clinical evidence of Lu Dangshen Oral Liquid and seven control drugs(Bufei Huoxue Capsules,Xiangsha Liujun Pills,Jinshuibao Capsules,Shenling Baizhu Granules,Yupingfeng Granules,Zhenqi Fuzheng Granules and Bailing Capsules)were obtained by using quantitative and qualitative methods through literature retrieval,questionnaire survey and data collection.Based on the multi criteria decision analysis(MCDA)model,the clinical value of drugs was comprehensively evaluated.The weights of criterion layer,index layer and alternative scheme were calculated by analytic hierarchy process.The equal frequency discretization rule was used to classify the evaluation results.Results The safety evidence showed that the main adverse reactions were epistaxis and nausea,and there were no serious adverse events.The comprehensive evaluation was grade A(the risk was controllable and the evidence was sufficient).Clinical studies showed that its treatment of lung spleen qi deficiency syndrome reflects the idea of treating different diseases with same method,and its efficacy in stable chronic obstructive pulmonary disease and children with recurrent respiratory tract infection was better than conventional treatment or Spleen Aminopeptide Oral Lyophilized Powder,and its effectiveness was rated as grade A.In terms of economy,the daily drug cost was relatively high,and it was rated as grade C.The preparation process was authorized by one patent.The single Chinese materia medica could improve the diseases and symptoms related to lung spleen qi deficiency syndrome,and the innovation was rated as grade A.In the questionnaire survey,the clinical use and patient compliance were excellent,and the suitability was rated as grade A.The supply of raw materials was stable,and the accessibility was rated as grade B.Conclusion The comprehensive evaluation of the clinical value of Lu Dangshen Oral Liquid in the treatment of lung spleen qi deficiency syndrome is grade A,which can provide references for the adjustment of the national medical insurance catalog and the national essential medicine catalog.

This study aims to isolate and culture primary rabbit spinal cord microvascular endothelial cells in vitro,providing a practical source of test cells for spinal cord injury research.Spinal cord tissue was aseptically extracted from one-month-old rabbits and processed sequentially through mincing,bovine serum albumin density gradient centrifugation,mesh filtration,and type Ⅱ collagenase digestion to ob-tain purified spinal cord microvascular segments.The microvascular segments were homogeneously mixed with an apprapriate volume of M199 complete culture medium and seeded into a culture dish for primary culture.Throughout the culture period,cell growth performance were continuously observed and recor-ded.Additionally,immunocytochemical staining was performed to evaluate the expression of factor Ⅷ-re-lated antigen.The results showed that after 24 hours of inoculation,a small amount of endothelial-like cells were observed to emerge from the spinal cord microvascular segments.Within 36～60 hours,the cell colonies gradually expanded and fused.After 72 hours,the cells spread across the base of the dish,forming a"cobblestone-like"monolayer.Immunocytochemical staining showed that more than 99％of the cells showed brown-red cytoplasm and were positive for factor Ⅷ-related antigen.It is these results that suggest this study has successfully established a convenient and stable primary rabbit spinal cord micro-vascular endothelial cells culture method.

Tuberculosis is still the second leading cause of death from a single source of infection in the world.There is a two-way relationship between tuberculosis and the nutritional status of the body,which affects and causes each other.Vitamin D is an essential micronutrient,most research results show that vitamin D deficiency is common in tuberculosis patients,which is related to lack of sunlight,decreased dietary intake of vitamin D and anti-tuberculosis drug treatment.Low level vitamin D can increase tuberculosis susceptibility to some extent,but the research results are not completely consistent.This paper reviews the nutritional status of vitamin D in tuberculosis patients in recent years,the causes of vitamin D deficiency in tuberculosis patients and the relationship between vitamin D deficiency and susceptibility of tuberculosis,so as to provide references for further study on the role of vitamin D in tuberculosis prevention and treatment.

Objective To explore the imaging and pathological features of pulmonary cryptococcosis(PC)in non-immunosuppressed patients.Methods A total of 56 non-immunosuppressed patients pathologically confirmed as PC were analyzed retrospectively,and the signs included halo sign,bronchial distension sign,superficial lobulation sign,and nodule accumulation,fusion sign were compared with the pathological features.Results CT images showed that the lesions of all 56 cases were located within 15 mm below the pleura.Distribution of the lesions were as follows:48 cases were located in unilateral lung(85.7％),8 cases in bilateral lungs(14.3％);47 cases were located in the lower lobes of the lungs(83.9％).Features of lesion morphology were as follows:48 cases were nodule/mass lesions(85.7％);6 cases were mixed lesions(10.7％);and 2 cases were solid infiltrates lesions(3.6％).Features of imaging signs were as follows:halo sign was in 46 cases(82.1％),bronchial distension sign in 31 cases(55.3％),superficial lobulation sign in 31 cases(55.3％),nodule accumulation,fusion sign in 26 cases(46.4％),burr sign in 14 cases(25.0％),cavity in 12 cases(21.4％);small amount of pleural effusion on the affected side in 5 cases(8.9％);and no enlarged lymph nodes were detected in the mediastinum.In 56 patients with PC,granulomatous inflammation pathologically characterized by multinucleated giant cells,epithelioid cell-like hyperplasia,and lymphocytic infiltration was predominantly identified in the nodule/mass lesions(45 cases),whereas pathologically non-granulomatous inflammation(lymphocytic infiltration/coagulative necrosis)was less identified.Conclusion In the non-immunosuppressed population,PC has more characteristic imaging and pathological manifestations,with predominantly nodule/mass on imaging,accompanied by halo sign,bronchial distension sign,superficial lobulation sign and nodule accumulation,fusion sign,etc.Pathologically,PC is mainly characterized by granulomatous inflammation,and the recognition of these signs is helpful in improving the conformity rate of the diagnostic imaging of this disease.

Aim To explore the mechanism of ligustil-ide(LIG)improving demyelination by inhibiting in-flammatory response in mice with distal middle cerebral artery occlusion(dMCAO)through AIM2/caspase-1 signal pathway.Methods Thirty C57BL/6N male mice were randomly divided into three groups:sham operation group(Sham group,n=10),model group(dMCAO group,n=10)and treatment group(LIG group,n=10).The dMCAO mouse model was estab-lished by electrocoagulation in dMCAO group and LIG group.The mice were scored by Longa after waking up,and the changes of cerebral blood flow were moni-tored by laser speckle blood flow imaging system after dMCAO.One hour after modeling,LIG(30 mg·kg-1·d-1)was injected intraperitoneally in LIG group,and the same amount of normal saline was injected in sham group and dMCAO group for one week until the end of the experiment.The mice in each group were stained with TTC,and the brain injury was observed pathologically.Fatigue turning bar test and open field test were used to evaluate the motor function and anxie-ty degree of mice,and then the brain tissues of mice were taken for black gold staining to compare the chan-ges of myelin sheath in each group.Immunofluores-cence staining was used to detect the average fluores-cence intensity of MBP,IBA1 and GFAP in CC,CPU and CX regions of mouse brains.ELISAwas used to de-tect the contents of TNF-α,IL-6,IL-1 β,IL-17A and BDNF in brain tissue proteins of mice.Western blot-ting was used to detect the protein expressions of AIM2,caspase-1 and ASC-in each group.Results Compared with the dMCAO group,the infarct area was reduced,the behavior was significantly improved and the demyelination was reduced in the LIG group.The expression of MBP protein in CC,CPU and CX regions increased(P＜0.05),the expression of IBA1 in CX decreased(P＜0.01),and the expression of GFAP in-creased in CC,CPU and CX regions(P＜0.01).The results of ELISA showed that the levels ofTNF-α(P＜0.01),IL-6(P＜0.01),IL-1β(P＜0.05)and IL-17A(P＜0.01)significantly decreased,while the ex-pression of BDNF increased(P＜0.05).The protein expression levels of AIM2,caspase-1 and ASC in mouse brain decreased after treatment(P＜0.01).Conclusion LIG has a protective effect on demyelina-tion in dMCAO mice,which may be related to the inhi-bition of AIM2/caspase-1 signaling pathway and in-flammation and to the promotion of BDNF secretion.

This study aims to isolate and culture primary rabbit spinal cord microvascular endothelial cells in vitro,providing a practical source of test cells for spinal cord injury research.Spinal cord tissue was aseptically extracted from one-month-old rabbits and processed sequentially through mincing,bovine serum albumin density gradient centrifugation,mesh filtration,and type Ⅱ collagenase digestion to ob-tain purified spinal cord microvascular segments.The microvascular segments were homogeneously mixed with an apprapriate volume of M199 complete culture medium and seeded into a culture dish for primary culture.Throughout the culture period,cell growth performance were continuously observed and recor-ded.Additionally,immunocytochemical staining was performed to evaluate the expression of factor Ⅷ-re-lated antigen.The results showed that after 24 hours of inoculation,a small amount of endothelial-like cells were observed to emerge from the spinal cord microvascular segments.Within 36～60 hours,the cell colonies gradually expanded and fused.After 72 hours,the cells spread across the base of the dish,forming a"cobblestone-like"monolayer.Immunocytochemical staining showed that more than 99％of the cells showed brown-red cytoplasm and were positive for factor Ⅷ-related antigen.It is these results that suggest this study has successfully established a convenient and stable primary rabbit spinal cord micro-vascular endothelial cells culture method.

Aim To explore the mechanism of ligustil-ide(LIG)improving demyelination by inhibiting in-flammatory response in mice with distal middle cerebral artery occlusion(dMCAO)through AIM2/caspase-1 signal pathway.Methods Thirty C57BL/6N male mice were randomly divided into three groups:sham operation group(Sham group,n=10),model group(dMCAO group,n=10)and treatment group(LIG group,n=10).The dMCAO mouse model was estab-lished by electrocoagulation in dMCAO group and LIG group.The mice were scored by Longa after waking up,and the changes of cerebral blood flow were moni-tored by laser speckle blood flow imaging system after dMCAO.One hour after modeling,LIG(30 mg·kg-1·d-1)was injected intraperitoneally in LIG group,and the same amount of normal saline was injected in sham group and dMCAO group for one week until the end of the experiment.The mice in each group were stained with TTC,and the brain injury was observed pathologically.Fatigue turning bar test and open field test were used to evaluate the motor function and anxie-ty degree of mice,and then the brain tissues of mice were taken for black gold staining to compare the chan-ges of myelin sheath in each group.Immunofluores-cence staining was used to detect the average fluores-cence intensity of MBP,IBA1 and GFAP in CC,CPU and CX regions of mouse brains.ELISAwas used to de-tect the contents of TNF-α,IL-6,IL-1 β,IL-17A and BDNF in brain tissue proteins of mice.Western blot-ting was used to detect the protein expressions of AIM2,caspase-1 and ASC-in each group.Results Compared with the dMCAO group,the infarct area was reduced,the behavior was significantly improved and the demyelination was reduced in the LIG group.The expression of MBP protein in CC,CPU and CX regions increased(P＜0.05),the expression of IBA1 in CX decreased(P＜0.01),and the expression of GFAP in-creased in CC,CPU and CX regions(P＜0.01).The results of ELISA showed that the levels ofTNF-α(P＜0.01),IL-6(P＜0.01),IL-1β(P＜0.05)and IL-17A(P＜0.01)significantly decreased,while the ex-pression of BDNF increased(P＜0.05).The protein expression levels of AIM2,caspase-1 and ASC in mouse brain decreased after treatment(P＜0.01).Conclusion LIG has a protective effect on demyelina-tion in dMCAO mice,which may be related to the inhi-bition of AIM2/caspase-1 signaling pathway and in-flammation and to the promotion of BDNF secretion.

Objective To explore the imaging and pathological features of pulmonary cryptococcosis(PC)in non-immunosuppressed patients.Methods A total of 56 non-immunosuppressed patients pathologically confirmed as PC were analyzed retrospectively,and the signs included halo sign,bronchial distension sign,superficial lobulation sign,and nodule accumulation,fusion sign were compared with the pathological features.Results CT images showed that the lesions of all 56 cases were located within 15 mm below the pleura.Distribution of the lesions were as follows:48 cases were located in unilateral lung(85.7％),8 cases in bilateral lungs(14.3％);47 cases were located in the lower lobes of the lungs(83.9％).Features of lesion morphology were as follows:48 cases were nodule/mass lesions(85.7％);6 cases were mixed lesions(10.7％);and 2 cases were solid infiltrates lesions(3.6％).Features of imaging signs were as follows:halo sign was in 46 cases(82.1％),bronchial distension sign in 31 cases(55.3％),superficial lobulation sign in 31 cases(55.3％),nodule accumulation,fusion sign in 26 cases(46.4％),burr sign in 14 cases(25.0％),cavity in 12 cases(21.4％);small amount of pleural effusion on the affected side in 5 cases(8.9％);and no enlarged lymph nodes were detected in the mediastinum.In 56 patients with PC,granulomatous inflammation pathologically characterized by multinucleated giant cells,epithelioid cell-like hyperplasia,and lymphocytic infiltration was predominantly identified in the nodule/mass lesions(45 cases),whereas pathologically non-granulomatous inflammation(lymphocytic infiltration/coagulative necrosis)was less identified.Conclusion In the non-immunosuppressed population,PC has more characteristic imaging and pathological manifestations,with predominantly nodule/mass on imaging,accompanied by halo sign,bronchial distension sign,superficial lobulation sign and nodule accumulation,fusion sign,etc.Pathologically,PC is mainly characterized by granulomatous inflammation,and the recognition of these signs is helpful in improving the conformity rate of the diagnostic imaging of this disease.

Objective To conduct a clinical comprehensive evaluation of Lu Dangshen Oral Liquid from 6 dimensions,including safety,effectiveness,economic value,innovativeness,suitability and accessibility.Methods The clinical evidence of Lu Dangshen Oral Liquid and seven control drugs(Bufei Huoxue Capsules,Xiangsha Liujun Pills,Jinshuibao Capsules,Shenling Baizhu Granules,Yupingfeng Granules,Zhenqi Fuzheng Granules and Bailing Capsules)were obtained by using quantitative and qualitative methods through literature retrieval,questionnaire survey and data collection.Based on the multi criteria decision analysis(MCDA)model,the clinical value of drugs was comprehensively evaluated.The weights of criterion layer,index layer and alternative scheme were calculated by analytic hierarchy process.The equal frequency discretization rule was used to classify the evaluation results.Results The safety evidence showed that the main adverse reactions were epistaxis and nausea,and there were no serious adverse events.The comprehensive evaluation was grade A(the risk was controllable and the evidence was sufficient).Clinical studies showed that its treatment of lung spleen qi deficiency syndrome reflects the idea of treating different diseases with same method,and its efficacy in stable chronic obstructive pulmonary disease and children with recurrent respiratory tract infection was better than conventional treatment or Spleen Aminopeptide Oral Lyophilized Powder,and its effectiveness was rated as grade A.In terms of economy,the daily drug cost was relatively high,and it was rated as grade C.The preparation process was authorized by one patent.The single Chinese materia medica could improve the diseases and symptoms related to lung spleen qi deficiency syndrome,and the innovation was rated as grade A.In the questionnaire survey,the clinical use and patient compliance were excellent,and the suitability was rated as grade A.The supply of raw materials was stable,and the accessibility was rated as grade B.Conclusion The comprehensive evaluation of the clinical value of Lu Dangshen Oral Liquid in the treatment of lung spleen qi deficiency syndrome is grade A,which can provide references for the adjustment of the national medical insurance catalog and the national essential medicine catalog.