Objective To investigate the neuroprotective effect of ligustilide(LIG)-mediated phosphatase and tensin homolog(PTEN)-induced putative kinase 1(PINK1)/Parkin pathway on mitophagy in rats with cerebral ischemia-reperfusion injury.Methods 161 male Sprague Dawley(SD)rats were randomly divided into sham operation(Sham)group,model group,LIG low-dose group,LIG high-dose group,mitophagy inhibitor(Mdivi-1)group,LIG high-dose+Mdivi-1 group,and the positive drug Nimodipine(NMDP)group,each with 23 rats.A modified middle cerebral artery wire thrombus method was used to construct a cerebral ischemia/reperfusion model in rats,and the neurobehavioral scores of rats in each group were compared by Longa's five-point scale;the volume of cerebral infarction was detected by 2,3,5-triphenyltetrazolium chloride(TCC)staining,the histopathology and ultrastructure of the hippocampus were examined by hematoxylin-eosin(HE)staining and transmission electron microscope(TEM).And the Na+-K+-Adenosine Triphosphate was measured by enzyme-linked immunosorbent assay(ELISA);double immunofluorescence staining for translocase of the outer membrane of mitochondrion 20(TOMM20)and Microtubule-associated protein 1 light chain 3(LC3)co-localized area percentage.Flow cytometry assay(FCM)to test the level of reactive oxygen(ROS);real-time fluorescence quantitative PCR(qRT-PCR)was used to measure the relative content of mitochondria in hippocampal neurons;and Western blot was performed to test the level of autophagy and the PINK1/Parkin pathway related protein expression.Results Compared with the Sham group,the neurological function score and cerebral infarction volume of the model group were increased,the hippocampal neurons showed pathological damage such as disordered arrangement,nucleolus disappearance and partial shrinkage of the nucleus and plasma,nuclear membrane rupture,swelling,membrane rupture and crista reduction of some mitochondria,a large number of autophagosomes were observed,and the colocalization area percentage of TOMM20 and LC3 was increased.TOMM20 and cytochrome C oxidase subunit IV isoform 1(COX4I1)in hippocampus and selective autophagy adaptor protein 62(p62)protein expression,mitochondrial encoded ATP synthase 6(mt-ATP6)/Ribosomal protein L13(Rpl13)ratio and Na+-K+-ATPase content decreased,while PINK1 and Parkin protein expression,LC3-II/I ratio and ROS relative content increased,and the differences were statistically significant(t=4.602～52.012,all P<0.01).Compared with the model group,the neurological function score,cerebral infarction volume,pathological and ultrastructural damage of hippocampal neurons were significantly improved in the LIG low,high dose and NMDP groups,and the differences were statistically significant(t=4.851～12.525,all P<0.01).The colocalization of TOMM20 and LC3 and the content of Na+-K+-ATPase were increased,while the expression of TOMM20,COX4I1 and p62 proteins and the mt-ATP6/Rpl13 ratio were decreased in the high-dose LIG group.The protein expression of PINK1 and Parkin,LC3-II/I ratio and ROS relative content were increased,and the differences were statistically significant(t=4.087～33.211,all P<0.01).Compared with the LIG high-dose group,the Mdivi-1 and LIG+Mdivi-1 groups had significantly decreased colocalization of TOMM20 and LC3 and Na+-K+-ATPase content,and significantly increased expression of TOMM20,COX4I1 and p62 proteins and mt-ATP6/Rpl13 ratio.The protein expression of PINK1 and Parkin,LC3-II/I ratio and ROS relative content were decreased,and the differences were statistically significant(t=4.008～43.415,all P<0.01).However,the percentage of TOMM20 and LC3 co-localization area,PINK1 and Parkin protein expression,LC3-II/I ratio and Na+-K+-ATPase content in the hippocampus of the LIG+Mdivi-1 group were higher than those of the Mdivi-1 group.The protein expression of COX4I1 and p62,mt-ATP6/Rpl13 ratio and ROS level were lower than those in MDIV-1 group,and the differences were statistically significant(t=3.721～21.513,all P<0.01).Conclusion LIG may activate mitophagy by regulating PINK1/Parkin signaling pathway to protect neurons from cerebral ischemia-reperfusion injury in rats.

Objective To investigate the neuroprotective effect of ligustilide(LIG)-mediated phosphatase and tensin homolog(PTEN)-induced putative kinase 1(PINK1)/Parkin pathway on mitophagy in rats with cerebral ischemia-reperfusion injury.Methods 161 male Sprague Dawley(SD)rats were randomly divided into sham operation(Sham)group,model group,LIG low-dose group,LIG high-dose group,mitophagy inhibitor(Mdivi-1)group,LIG high-dose+Mdivi-1 group,and the positive drug Nimodipine(NMDP)group,each with 23 rats.A modified middle cerebral artery wire thrombus method was used to construct a cerebral ischemia/reperfusion model in rats,and the neurobehavioral scores of rats in each group were compared by Longa's five-point scale;the volume of cerebral infarction was detected by 2,3,5-triphenyltetrazolium chloride(TCC)staining,the histopathology and ultrastructure of the hippocampus were examined by hematoxylin-eosin(HE)staining and transmission electron microscope(TEM).And the Na+-K+-Adenosine Triphosphate was measured by enzyme-linked immunosorbent assay(ELISA);double immunofluorescence staining for translocase of the outer membrane of mitochondrion 20(TOMM20)and Microtubule-associated protein 1 light chain 3(LC3)co-localized area percentage.Flow cytometry assay(FCM)to test the level of reactive oxygen(ROS);real-time fluorescence quantitative PCR(qRT-PCR)was used to measure the relative content of mitochondria in hippocampal neurons;and Western blot was performed to test the level of autophagy and the PINK1/Parkin pathway related protein expression.Results Compared with the Sham group,the neurological function score and cerebral infarction volume of the model group were increased,the hippocampal neurons showed pathological damage such as disordered arrangement,nucleolus disappearance and partial shrinkage of the nucleus and plasma,nuclear membrane rupture,swelling,membrane rupture and crista reduction of some mitochondria,a large number of autophagosomes were observed,and the colocalization area percentage of TOMM20 and LC3 was increased.TOMM20 and cytochrome C oxidase subunit IV isoform 1(COX4I1)in hippocampus and selective autophagy adaptor protein 62(p62)protein expression,mitochondrial encoded ATP synthase 6(mt-ATP6)/Ribosomal protein L13(Rpl13)ratio and Na+-K+-ATPase content decreased,while PINK1 and Parkin protein expression,LC3-II/I ratio and ROS relative content increased,and the differences were statistically significant(t=4.602～52.012,all P<0.01).Compared with the model group,the neurological function score,cerebral infarction volume,pathological and ultrastructural damage of hippocampal neurons were significantly improved in the LIG low,high dose and NMDP groups,and the differences were statistically significant(t=4.851～12.525,all P<0.01).The colocalization of TOMM20 and LC3 and the content of Na+-K+-ATPase were increased,while the expression of TOMM20,COX4I1 and p62 proteins and the mt-ATP6/Rpl13 ratio were decreased in the high-dose LIG group.The protein expression of PINK1 and Parkin,LC3-II/I ratio and ROS relative content were increased,and the differences were statistically significant(t=4.087～33.211,all P<0.01).Compared with the LIG high-dose group,the Mdivi-1 and LIG+Mdivi-1 groups had significantly decreased colocalization of TOMM20 and LC3 and Na+-K+-ATPase content,and significantly increased expression of TOMM20,COX4I1 and p62 proteins and mt-ATP6/Rpl13 ratio.The protein expression of PINK1 and Parkin,LC3-II/I ratio and ROS relative content were decreased,and the differences were statistically significant(t=4.008～43.415,all P<0.01).However,the percentage of TOMM20 and LC3 co-localization area,PINK1 and Parkin protein expression,LC3-II/I ratio and Na+-K+-ATPase content in the hippocampus of the LIG+Mdivi-1 group were higher than those of the Mdivi-1 group.The protein expression of COX4I1 and p62,mt-ATP6/Rpl13 ratio and ROS level were lower than those in MDIV-1 group,and the differences were statistically significant(t=3.721～21.513,all P<0.01).Conclusion LIG may activate mitophagy by regulating PINK1/Parkin signaling pathway to protect neurons from cerebral ischemia-reperfusion injury in rats.

[Objective]To study the formation,change and propagation of Liuwei Dihuang Pill.[Methods]The medical literature and other historical records about Liuwei Dihuang Pill were collected and analyzed,and the formation process,effect change,influencing factors behind the change and application of this prescription and the transmission path were discussed.[Results]Liuwei Dihuang Pill was developed by QIAN Yi,a doctor in Song Dynasty,based on the decoction of Shengqi Pill.It was used to treat kidney deficiency syndrome in children.Because the cooked Rehmannia can nourish Yin and blood,Song and Yuan Dynasties doctors also used the prescription for Yin blood deficiency.In the Ming Dynasty,with the elucidations of the Mingmen theory by the doctors of the warm-tonifying faction,this prescription became the representative formula for tonifying kidney Yin.At the same time,the spreading and population of this prescription was not only in the elite medical group,but also through the medical heritage of GAO Doukui,LYU Liuliang and FANG Yizhi,in the form of medical books,prescription books and health books in the folk.By the Qing Dynasty,the spreading of this formula had shown the characteristics of"secularization",and the literature,literati's notes,novels and other materials can reflect the use of Liuwei Dihuang Pill in the folk.[Conclusion]This paper discusses the origin and development of Liuwei Dihuang Pill,which is of great significance for the study of the rise and formation of the warm-tonifying thought in the Ming and Qing Dynasties.

[Objective]To study the formation,change and propagation of Liuwei Dihuang Pill.[Methods]The medical literature and other historical records about Liuwei Dihuang Pill were collected and analyzed,and the formation process,effect change,influencing factors behind the change and application of this prescription and the transmission path were discussed.[Results]Liuwei Dihuang Pill was developed by QIAN Yi,a doctor in Song Dynasty,based on the decoction of Shengqi Pill.It was used to treat kidney deficiency syndrome in children.Because the cooked Rehmannia can nourish Yin and blood,Song and Yuan Dynasties doctors also used the prescription for Yin blood deficiency.In the Ming Dynasty,with the elucidations of the Mingmen theory by the doctors of the warm-tonifying faction,this prescription became the representative formula for tonifying kidney Yin.At the same time,the spreading and population of this prescription was not only in the elite medical group,but also through the medical heritage of GAO Doukui,LYU Liuliang and FANG Yizhi,in the form of medical books,prescription books and health books in the folk.By the Qing Dynasty,the spreading of this formula had shown the characteristics of"secularization",and the literature,literati's notes,novels and other materials can reflect the use of Liuwei Dihuang Pill in the folk.[Conclusion]This paper discusses the origin and development of Liuwei Dihuang Pill,which is of great significance for the study of the rise and formation of the warm-tonifying thought in the Ming and Qing Dynasties.

Objective: To investigate the prognostic utility of radiomics features extracted from 18 F-fluorodeoxyglucose (FDG) PET/CT combined with clinical factors and metabolic parameters in predicting progression-free survival (PFS) and overall survival (OS) in individuals diagnosed with extranodal nasal-type NK/T cell lymphoma (ENKTCL). Materials and Methods: A total of 126 adults with ENKTCL who underwent 18 F-FDG PET/CT examination before treatment were retrospectively included and randomly divided into training (n = 88) and validation cohorts (n = 38) at a ratio of 7:3.Least absolute shrinkage and selection operation Cox regression analysis was used to select the best radiomics features and calculate each patient’s radiomics scores (RadPFS and RadOS). Kaplan–Meier curve and Log-rank test were used to compare survival between patient groups risk-stratified by the radiomics scores. Various models to predict PFS and OS were constructed, including clinical, metabolic, clinical + metabolic, and clinical + metabolic + radiomics models. The discriminative ability of each model was evaluated using Harrell’s C index. The performance of each model in predicting PFS and OS for 1-, 3-, and 5-years was evaluated using the time-dependent receiver operating characteristic (ROC) curve. Results: Kaplan–Meier curve analysis demonstrated that the radiomics scores effectively identified high- and low-risk patients (all P < 0.05). Multivariable Cox analysis showed that the Ann Arbor stage, maximum standardized uptake value (SUVmax), and RadPFS were independent risk factors associated with PFS. Further, β2-microglobulin, Eastern Cooperative Oncology Group performance status score, SUVmax, and RadOS were independent risk factors for OS. The clinical + metabolic + radiomics model exhibited the greatest discriminative ability for both PFS (Harrell’s C-index: 0.805 in the validation cohort) and OS (Harrell’s C-index: 0.833 in the validation cohort). The time-dependent ROC analysis indicated that the clinical + metabolic + radiomics model had the best predictive performance. Conclusion The PET/CT-based clinical + metabolic + radiomics model can enhance prognostication among patients with ENKTCL and may be a non-invasive and efficient risk stratification tool for clinical practice.

Objective To investigate the current situation of Staphylococcus aureus contamination in raw and cooked meat and dairy products in Wuhan, and analyze the enterotoxins production and antimicrobial resistance of isolated bacterial strains. Methods The detection of Staphylococcus aureus was performed according to GB4789.10-2016 National Food Safety Standard. Staphylococcus aureus enterotoxin (SE) PCR kit and ELISA were used for SEA-E type detection. Broth dilution and PCR method were used for drug sensitivity test. Results A total of 13 strains of Staphylococcus aureus were isolated from 202 samples, and the isolation rate of Staphylococcus aureus in the raw and cooked meat and dairy products was 6.43%. The detection rate of Staphylococcus aureus was 9.82% (11/112) in raw meat, and 4% (2/50) in cooked meat products. There was no detection in dairy products. Of the 13 isolated strains, 6 strains were found to have enterotoxins, with a toxin production rate of 46.15% (6/13). Among the 6 strains of enterotoxin producing Staphylococcus aureus, 4 strains were classified as type A, C, D, and AB, respectively. The isolated strains were generally resistant to tetracycline and sulfonamide drugs, and the detection rate of resistant genes was more than 60%. The resistance rate to penicillin and erythromycin exceeded 50%, and the dominant resistance spectrum was the detection of 3 strains of single-resistant (PEN) Staphylococcus aureus (25.08%, 3/13), followed by 2 strains of five-fold resistance (PEN-ERY-CLI-SXT-GEN), and double resistance Staphylococcus aureus (PEN-ERY) (15.38%, 2/13). Genetic testing was consistent with phenotypic testing. Conclusion In 2020, there was a certain degree of contamination of Staphylococcus aureus in raw and cooked meat products in Wuhan, with 13 isolated strains and 6 strains producing enterotoxins. It is necessary to remain vigilant about the potential food risks of raw and cooked meat products, and strengthen the supervision of the safety risks of raw and cooked meat products.

Objective: To evaluate the immunity and influencing factors of diphtheria among preschool children in Shenzhen,to provide reference for effective monitoring of diphtheria IgG antibody level in preschool children. Methods: Serum samples were collected from 296 preschool children aged 4-6 who were recruited in Shenzhen. The diphtheria antibody titer in serum was determined by enzyme linked immunosorbent assay, and the effect of different immumuzation schedule including types of vaccine and vaccination timing, on the geometric mean concentration (GMC) of diphtheria IgG antibody and antibody positive rate were analyzed. Results: The GMC of diphtheria IgG antibody was 0.71 IU/mL, and the positive conversion rate was 33.1%. There were significant differences in antibody GMC and antibody positive conversion rate of diphtheria in different age groups( F/χ 2=11.77, 27.45, P < 0.01 ). The GMC and antibody positive conversion rate showed significant differences by diphtheria antibodies, vaccine types and end dose vaccination intervals( F=49.53, 12.95，11.61, P <0.01). There were statistically significant differences in the positive conversion rate of diphtheria antibodies in children with different types of diphtheria antibodies, vaccine types of diphtheria antibodies, and diphtheria antibodies at the time interval of final vaccination (Fisher exact probability method, P <0.01). Conclusion The overall positive conversion rate of diphtheria antibody in preschool children in Shenzhen is high. Timely completion of full diphtheria vaccination can improve the antibody level and plays a better role in protecting preschool children.

Although widely applied in treating hematopoietic malignancies, transplantation of hematopoietic stem/progenitor cells (HSPCs) is impeded by HSPC shortage. Whether circulating HSPCs (cHSPCs) in steady-state blood could be used as an alternative source remains largely elusive. Here we develop a three-dimensional culture system (3DCS) including arginine, glycine, aspartate, and a series of factors. Fourteen-day culture of peripheral blood mononuclear cells (PBMNCs) in 3DCS led to 125- and 70-fold increase of the frequency and number of CD34⁺ cells. Further, 3DCS-expanded cHSPCs exhibited the similar reconstitution rate compared to CD34⁺ HSPCs in bone marrow. Mechanistically, 3DCS fabricated an immunomodulatory niche, secreting cytokines as TNF to support cHSPC survival and proliferation. Finally, 3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization. Our 3DCS successfully expands rare cHSPCs, providing an alternative source for the HSPC therapy, particularly for the patients/donors who have failed in HSPC mobilization.
Antigens, CD34/metabolism* ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Leukocytes, Mononuclear/metabolism* ; Peptides/metabolism*

Long non-coding RNA (lncRNA) contains rich information and functions. The research on Traditional Chinese Medicine (TCM) targeting lncRNA mainly involves tumors, cardiovascular diseases, endocrine and metabolic related diseases, osteoporosis and other diseases, which are used to explore the mechanism of TCM and the differetiation of TCM syndromes or constitution, etc. LncRNA has important application prospects in the field of TCM. The study of lncRNA may provide new ways and technical methods for the research of modern TCM.

Objective::Microchannels are associated with the progression of atherosclerotic vulnerable plaques. However, in patients with culprit optical coherence tomography (OCT)-defined plaque erosion, the knowledge of microchannels and culprit lesion vulnerability is limited. The aim of this study was to investigate culprit lesion characteristics in patients with ST-segment elevated myocardial infarction (STEMI) caused by plaque erosion with and without microchannels using OCT.Methods::In all, 348 STEMI patients with plaque erosion who underwent OCT of the culprit lesion at the 2 nd Affiliated Hospital of Harbin Medical University (Harbin, China) from August 2014 to December 2017 were included and divided into the microchannel group ( n= 116, 33.3%) and no-microchannel group ( n = 232, 66.7%). The clinical characteristics and OCT-derived plaque features were compared between both groups. Results::Among the 348 STEMI patients with plaque erosion, culprit lesions with microchannels had higher incidence of lipid plaque (59.5% vs. 45.3%, P= 0.012); calcification (41.4% vs. 24.6%, P= 0.002); spotty calcification (30.2% vs. 18.1%, P= 0.014); macrophages accumulation (72.4% vs. 45.7%, P < 0.001); and cholesterol crystals (32.8% vs. 14.2%, P < 0.001) than those without microchannels. In addition, minimal lumen area was smaller ((1.9 ± 0.9) mm 2vs. (2.8 ± 2.3) mm 2, P < 0.001) and lumen area stenosis was greater ((71.3% ± 13.4%) vs. (65.3% ± 19.3%), P= 0.001) in the microchannel group than in the no-microchannel group. Conclusion::In patients with STEMI caused by plaque erosion, one-third manifested typical microchannel characteristics, and those with microchannels were associated with more severe luminal stenosis and more vulnerable plaque features than those without microchannels.