ObjectiveTo investigate the molecular mechanisms of Mahuang Fuzi Gancao Tang and its pungent single herbs， Ephedrae Herba and Aconiti Lateralis Radix Praeparaia， in repairing podocyte injury based on the sweat pore theory， with a focus on the podocyte cytoskeletal transient receptor potential canonical 5 （TRPC5）-Ras-related C3 botulinum toxin substrate 1 （RAC1） feedforward loop. MethodsAn animal model with puromycin aminonucleoside （PAN）-induced overexpression of TRPC5 was established. Interventions included Mahuang Fuzi Gancao Tang， Ephedrae Herba alone， and Aconiti Lateralis Radix Praeparaia alone. Biochemical parameters ， histopathological changes， and podocyte ultrastructure were analyzed. Western blotting was performed to determine the expression of cytoskeletal protein synaptopodin and mechanism-related proteins TRPC5， RAC1-GTP， and RAC1 in the kidney. Primary podocytes were isolated and cultured for three-dimensional imaging of foot processes， cytoskeletal fluorescence analysis， and TRPC5-RAC1 co-staining via immunofluorescence. ResultsCompared with the model group， Mahuang Fuzi Gancao Tang， Ephedrae Herba alone， and Aconiti Lateralis Radix Praeparaia alone increased serum albumin （ALB）， decreased UPCR， reduced podocyte foot process fusion rate， upregulated synaptopodin expression， and downregulated TRPC5， RAC1-GTP， and RAC1 expression （P<0.05）. Moreover， the interventions increased the phalloidin fluorescence area/field area ratio （P<0.01） and mean fluorescence intensity （P<0.05）， while decreasing the proportion of TRPC5-RAC1 co-stained double-positive cells/total cells per field （P<0.01） in primary podocytes. ConclusionMahuang Fuzi Gancao Tang and its pungent single herbs， Ephedrae Herba and Aconiti Lateralis Radix Praeparaia， ameliorated podocyte injury in the model with PAN-induced TRPC5 overexpression by reducing proteinuria and suppressing the TRPC5-RAC1 feedforward loop-mediated podocyte cytoskeletal damage.

ObjectiveTo investigate the molecular mechanisms of Mahuang Fuzi Gancao Tang and its pungent single herbs， Ephedrae Herba and Aconiti Lateralis Radix Praeparaia， in repairing podocyte injury based on the sweat pore theory， with a focus on the podocyte cytoskeletal transient receptor potential canonical 5 （TRPC5）-Ras-related C3 botulinum toxin substrate 1 （RAC1） feedforward loop. MethodsAn animal model with puromycin aminonucleoside （PAN）-induced overexpression of TRPC5 was established. Interventions included Mahuang Fuzi Gancao Tang， Ephedrae Herba alone， and Aconiti Lateralis Radix Praeparaia alone. Biochemical parameters ， histopathological changes， and podocyte ultrastructure were analyzed. Western blotting was performed to determine the expression of cytoskeletal protein synaptopodin and mechanism-related proteins TRPC5， RAC1-GTP， and RAC1 in the kidney. Primary podocytes were isolated and cultured for three-dimensional imaging of foot processes， cytoskeletal fluorescence analysis， and TRPC5-RAC1 co-staining via immunofluorescence. ResultsCompared with the model group， Mahuang Fuzi Gancao Tang， Ephedrae Herba alone， and Aconiti Lateralis Radix Praeparaia alone increased serum albumin （ALB）， decreased UPCR， reduced podocyte foot process fusion rate， upregulated synaptopodin expression， and downregulated TRPC5， RAC1-GTP， and RAC1 expression （P<0.05）. Moreover， the interventions increased the phalloidin fluorescence area/field area ratio （P<0.01） and mean fluorescence intensity （P<0.05）， while decreasing the proportion of TRPC5-RAC1 co-stained double-positive cells/total cells per field （P<0.01） in primary podocytes. ConclusionMahuang Fuzi Gancao Tang and its pungent single herbs， Ephedrae Herba and Aconiti Lateralis Radix Praeparaia， ameliorated podocyte injury in the model with PAN-induced TRPC5 overexpression by reducing proteinuria and suppressing the TRPC5-RAC1 feedforward loop-mediated podocyte cytoskeletal damage.

[Objective]To discuss the pathogenesis,hierarchy,treatment and medication rules of deficiency of Yang with hyperactivity of Yin syndrome in Treatise on Febrile Diseases.[Methods]Combining with the medical theory of later physicians,this paper clarifies the pathogenesis and syndrome of deficiency of Yang with hyperactivity of Yin syndrome,explains the mechanism of Yang deficiency and Yin excess syndrome from the perspective of pathophysiology.This paper also analyzes the differences among the various versions of the text of the Shaoyin Chapter including the variations in disease location,the pathogen nature,stomach Qi and body fluid,and inducts the implicit therapeutic principles in the Shaoyin Chapter.[Results]The principle of differentiation and treatment for deficiency of Yang with hyperactivity of Yin syndrome is"protecting stomach Qi and preserving body fluids",which can be subdivided into Taiyin damp earth disease,Shaoyin monarch fire disease,and Jueyin wind wood disease.Through careful observation,questioning,smelling and palpation,combined with the pathological and physiological basis of modern medicine,it can accurately diagnose deficiency of Yang with hyperactivity of Yin syndrome.Shaoyin plays a crucial role in Treatise on Febrile Diseases.The structure of the Shaoyin Chapter is well-organized and can dynamically reflect the changes in the pathogenesis of Shaoyin syndrome,the combination of Shaoyin and Taiyin or Jueyin,or the tri-coordination syndrome of these location.From the text,it can be inferred that the location of the disease begins on the surface and gradually extends to the interior.The treatment methods will include moxibustion,warming the meridians,warming Yang and dredging Yang.The severity of the illness and the presence of phlegm-fluid determine the treatment,which may involve dispersing cold,dispersing fluids,and promoting urination.The degree of exhaustion of bodily fluids may require either reviving Yang for resuscitation or nourishing Yin.[Conclusion]From a pathological and physiological perspective,the mechanisms behind the changes in the primary symptoms of traditional Chinese medicine can be explained.When treating deficiency of Yang with hyperactivity of Yin syndrome,the selection of herbal prescriptions should be based on factors such as the disease location,the pathogen nature,stomach Qi and body fluids.The treatment principles for deficiency of Yang with hyperactivity of Yin syndrome have instructive significance for the treatment of other syndromes as well.

Objective The plasma von Willebrand factor(vWF)level in patients with primary microvascular angina(PMVA)were measured to evaluate the vascular endothelial function of the patients.The change of vWF level in patients after the treatment with Shexiangbaoxin Pill were observeg.Methods Totally 69 patients who were definitely diagnosed as PMVA,They were randomly divided into conventional treatment group(33cases)and ShexiangBaoxin Pill group(36cases).The plasma vWF levels of the two groups were measured before and after treatment.Results The level of vWF before treatment in conventional treatment group was(50.93±32.98)μg/L.The level of vWF before treatment in ShexiangBaoxin Pill group was(27.45±25.02)μg/L.The level of vWF in conventional treatment group after treatment was(49.65±35.12)μg/L.The level of vWF after treatment in ShexiangBaoxin Pill group was(17.37±15.68)μg/L.The difference of vWF decrease in Baoxin Pill group after treatment(10.08±16.47)μg/L,was lower than that in conventional treatment group(1.28±12.37)μg/L,the difference is significant(P<0.05).Conclusion Shexiang Baoxin Pill has the function of protecting vascular endothelium,and PMVA patients can benefit from treatment.

The"lipid nephrotoxicity"hypothesis introduces the lipid metabolism disorders into the field of kidney disease research.Fatty acids as important components of lipids can be ectopic deposition in various cells of the kidney in pathological status,and are involved in the occurrence and progression of acute kidney in-jury(AKI)and chronic kidney disease(CKD).The mechanism of abnormal fatty acid metabolism in tubular epithelial cells(TEC)and its function change are closely associated with kidney injury.This article summa-rized the relationship between fatty acid metabolism in TEC with AKI and CKD,elaborated the specific mech-anism of fatty acid metabolism disorders leading to TEC injury,and discussed the treatment strategies based on regulating fatty acid metabolism disorders to improve kidney injury.

Objective:To analyze levels of oral Streptococcus sanguinis( Ss)in middle-aged and elderly patients with primary microvascular angina(PMVA)and changes in vascular endothelial function. Methods:In this case-control study, 21 middle-aged and elderly patients diagnosed with PMVA at the Department of Cardiology, Hangzhou Red Cross Hospital between January 2019 and July 2022(the PMVA group)were recruited, with ages ranging from 45 to 80(63.4±12.3)years, while 23 healthy individuals receiving health checkups during the same period served as the control group, with ages ranging from 48-76(62.5±6.5)years.The 21 middle-aged and elderly PMVA patients underwent tests for the measurement of subgingival plaque Ss levels of the oral cavity and levels of plasma vascular von Willebrand factor(VWF)and homocysteine(Hcy). Pearson linear regression analysis was conducted.Results:Ss was not found in subgingival plaque of the oral cavity in the control group, but low levels of Ss were detected in patients from the PMVA group(percentage: 1.754×10 -4; 6.218×10 -5, 4.450×10 -4). The VWF level in the PMVA group was higher than in the control group[(20.22 ± 4.44)μg/L vs.(12.00 ± 6.60)μg/L, t=4.890, P<0.01]. There was no statistical difference in the Hcy level between the PMVA group and the control group[(15.28±6.40)μmol/L vs.(12.86±2.63)μmol/L, t=1.615, P>0.05]. There was no significant correlation between Ss levels and VWF levels in the PMVA group( r=0.038, P>0.05). Conclusions:Ss can be detected in subgingival plaque of the oral cavity in PMVA patients, but not in healthy middle-aged and elderly people.The VWF level in PMVA patients is significantly higher than in healthy people, indicating that vascular endothelial function is impaired in middle-aged and elderly PMVA patients.However, there is no correlation between subgingival plaque Ss levels of the oral cavity and VWF levels in PMVA patients.

Objective:To investigate plasma levels of fibrinopeptide A(FPA)and platelet alpha-granule membrane glycoprotein 140(GMP-140), and to analyze the changes of coagulation activity in elderly patients with primary microvascular angina(PMVA).Methods:In this prospective case-control study, a total of 45 elderly patients(aged 60-80 years, 27 males and 18 females)admitted to our hospital from Jan.2019 to Dec.2020 were diagnosed as PMVA(as microvascular angina group)by clinical symptoms, electrocardiogram, dynamic electrocardiogram and coronary angiography.Forty-three age-and sex-matched elderly subjects who took a medical check-up and had no primary microvascular angina were included as the control group.Plasma levels of FPA, GMP-140 and high-sensitivity C-reactive protein were determined.The levels of systolic blood pressure, diastolic blood pressure, platelet, fasting blood glucose, triglyceride, total cholesterol, high-density lipoprotein and low-density lipoprotein, uric acid, renal function and liver function were detected.Results:The FPA level was higher in the PMVA group than in the control group[(430.32±364.05)μg/L vs.(263.73± 118.29)μg/L, t=2.913, P<0.01]. There was no difference in GMP-140 level between the PMVA group and the control group[(5.78±3.92)μg/L vs.(6.95±1.91)μg/L, t=-1.790, P>0.05]. The high-sensitivity C-reactive protein level was higher in the PMVA group than in the control group[(3.33±5.70)mg/L vs.(0.81±0.86)mg/L, t=2.927, P<0.01]. Conclusions:Compared with the control group, the FPA level is significantly increased with elevated coagulation activity in patients with primary microvascular angina.The increased inflammatory state in the PMVA group might promote the occurrence and development of microvascular angina, but whether the inflammatory state promotes the enhancement of coagulation activity needs further study to confirm.

The emergence of regular short repetitive palindromic sequence clusters (CRISPR) and CRISPR- associated proteins 9 (Cas9) gene editing technology has greatly promoted the wide application of genetically modified pigs. Efficient single guide RNA (sgRNA) is the key to the success of gene editing using CRISPR/Cas9 technology. For large animals with a long reproductive cycle, such as pigs, it is necessary to screen out efficient sgRNA
Animals ; CRISPR-Cas Systems/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; Gene Editing ; RNA, Guide/genetics* ; Swine

Objective: To investigate the effects of antimicrobial peptides merecidin on the biological functions of human lung adenocarcinomaA549 cells and the potential signaling pathways and targets that involved in promoting apoptosis, by studying the changes of phosphorylation levels of proteins in A549 cells after merecidin treatment. Methods: The antibacterial peptide mericidin (9 μmol/L) was applied to treat A549 cells for 6 h, and the total protein was collected and extracted. The peptide was digested by trypsin and labeled with TMT, and then fractionated by HPLC. The phosphorylated peptides were enriched with IMAC-Fe, and finally subjected to mass spectrometry analysis. Library identification and quantification of phosphorylated peptides obtained by mass spectrometry were processed using MaxQuant software, to further analyze the functions and pathways of differentially expressed phosphorylated proteins by combining with bioinformatic analysis. Results: Through IPA analysis of phosphorylated proteins in the normal control group and the antibacterial peptide merecidin treatment group, 753 differentially phosphorylated proteins in mericidin treatment group were screened out under the conditions of |Fold Change|≥2 and P<0.05, including 229 significantly up-regulated genes and 417 down-regulated genes. Among them, the differentially expressed proteins related to apoptosis included RB1, MAPK1, ARAF, PTK2, FOXO, MARCKSandsoon.Theresultsofbiologicalprocessanalysisshowedthatdifferentiallyexpressed phosphorylated proteins were mainly concentrated in cell signal transduction, degradation and transport of nucleic acid, and cellular energy metabolism, protein translation and synthesis, and cytoskeleton formation etc. The enrichment results showed that the differentially expressed phosphorylated proteins were mainly involved in apoptosis-related MAPK, ErbB, PI3K-Akt, and Ras signaling pathways. Protein-protein interaction analysis indicated the associations among apoptosis-related proteins PTK2, PRKCA, MA2PK2, MAPK1, and LMNA. Conclusion: The antibacterial peptide merecidin may induce apoptosis and alteration of other cell functions by affecting a variety of genes and signaling pathways such as RB1, MAPK1,ARAF, PTK2, FOXO and MARCKS etc.

Objective:To analyze the mutations in desmosomal protein genes of arrhythmogenic right ventricular cardiomyopathy (ARVC) in relatives of Yunnan unexplained sudden death (YUSD) cases in Nanjian County, Dali Prefecture, Yunnan Province, and provide a basis for etiological hypothesis and control measures.Methods:The blood samples of YUSD case relatives ( n = 7) and control villagers ( n = 7) were collected, and basic situation investigation and electrocardiography (ECG) examination were performed at the same time. Blood DNA was extracted as a template for PCR amplification, and Sanger method was used to perform plakophilin 2 (PKP2), desmoglein 2 (DSG2), desmocollin 2 (DSC2), desmoplakin (DSP), and junction plakoglobin (JUP) five ARVC desmosomal protein genes sequencing of a total of 97 exons, and comprehensive analysis of gene mutations was carried out. Results:Five of YUSD case relatives carried genetic mutation sites, including DSP gene heterozygous synonymous mutations about exon 20 c.2862 C>T (p.Cys954Cys) and exon 24F c.7122 C>T (p.Thr2374Thr), DSC2 gene heterozygous missense mutation about exon 15 c.2326 A>G (p.Ile776Val), and all the five people were single heterozygous mutation carriers. Among them, two case relatives of the father-son carried the same site mutation of the DSC2 gene; the abnormal ECGs of three YUSD case relatives were ST-T change or clockwise rotation. However, the mutation sites of PKP2, DSG2, DSC2, DSP and JUP genes in control villagers were not detected.Conclusions:YUSD case relatives in Nanjian County carry ARVC desmosomal protein genes DSP and DSC2 mutations. Pathogenic mutation of DSC2 gene c.2326 A>G (p.Ile776Val) is may related to the incidence of some YUSD cases.