This paper summarizes professor LI Guolie's clinical experience in treating orthostatic hypotension （OH） based on the theory of "raising the clear and directing the turbid downward". It is considered that the core pathogenesis of OH lies in the body's transition from a supine to an upright position， during which dysfunction of the middle jiao （焦） transformation and transportation， along with impaired pivot function， hinders the ascending of clear yang and the descending of turbid yin. Treatment should follow the general principle of "ascending the clear and directing the turbid downward"， placing emphasis on distinguishing the primary and secondary aspects. For cases where the clear yang fails to ascend， the self-formulated Li's Shengqing Jiangzhuo Decoction （李氏升清降浊汤）is used to supplement qi， raise the clear， and strengthen the middle jiao. For cases where the turbid yin fails to descend， the self-formulated Wuxiang Qingzhuo Beverage（五香清浊饮）with modifications is applied to resolve phlegm， eliminate stasis， harmonize the middle， and descend the turbid.

This paper summarizes professor LI Guolie's clinical experience in treating orthostatic hypotension （OH） based on the theory of "raising the clear and directing the turbid downward". It is considered that the core pathogenesis of OH lies in the body's transition from a supine to an upright position， during which dysfunction of the middle jiao （焦） transformation and transportation， along with impaired pivot function， hinders the ascending of clear yang and the descending of turbid yin. Treatment should follow the general principle of "ascending the clear and directing the turbid downward"， placing emphasis on distinguishing the primary and secondary aspects. For cases where the clear yang fails to ascend， the self-formulated Li's Shengqing Jiangzhuo Decoction （李氏升清降浊汤）is used to supplement qi， raise the clear， and strengthen the middle jiao. For cases where the turbid yin fails to descend， the self-formulated Wuxiang Qingzhuo Beverage（五香清浊饮）with modifications is applied to resolve phlegm， eliminate stasis， harmonize the middle， and descend the turbid.

ObjectiveThis study aims to explore the mechanism of action of Huangqi Guizhi Wuwutang in treating rheumatoid arthritis （RA）-associated sarcopenia by regulating autophagy and improving skeletal muscle homeostasis based on network pharmacology，bioinformatics，machine learning，and animal experiments. MethodsActive ingredients and targets of Huangqi Guizhi Wuwutang were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP），PubChem，and SwissTargetPrediction databases. RA-related datasets were retrieved from the GEO database，and differential genes were screened. Sarcopenia-related targets were searched through GeneCards and the Comparative Toxicology Database （CTD），and autophagy-related gene sets were downloaded from the Human Autophagy Database （HADb）. Their intersection was analyzed to identify autophagy-related therapeutic targets，followed by enrichment analysis. A protein-protein interaction （PPI） network was constructed using the STRING database，and key targets were selected using multiple methods. Machine learning was applied to predict models based on the expression profiles of intersecting targets，and nomogram models were constructed based on key targets. Molecular docking of the top four active ingredients with key targets was performed using AutoDockVina. A collagen-induced arthritis （CIA） rat model was established using bovine type Ⅱ collagen，with SD rats divided into groups including a blank group，a model group，and low-，medium-，and high-dose groups of Huangqi Guizhi Wuwutang （2.44，4.88，and 9.76 g·kg^-1） and administered for five consecutive weeks. Joint scores and gastrocnemius muscle mass were recorded and analyzed after modeling. Hematoxylin and eosin （HE） staining and Masson's staining were used to observe pathological changes in muscle tissue. Immunofluorescence staining was applied to observe the protein expression levels of myosin heavy chain （MYHC） and insulin-like growth factor-1 （IGF-1） in skeletal muscle. Western blot was used to detect the protein expression levels of autophagy-related proteins ATG5，Beclin1，LC3B，muscle-specific proteins （MuRF1），MaFbx，and MYHC. Real-time quantitative reverse transcription PCR （Real-time PCR） was performed to measure the mRNA expression levels of ATG5，Beclin1，LC3B，MuRF1，MaFbx，and MYHC in muscle tissue. ResultsNetwork pharmacology revealed that Huangqi Guizhi Wuwutang shared 25 common targets with autophagy genes related to RA-associated sarcopenia. The PPI network and machine learning identified six key targets，which were primarily involved in autophagy and inflammatory pathways. Animal experiments showed that compared to the blank group，the model group had significantly higher joint scores （P<0.01） and lower gastrocnemius muscle index （P<0.01）. HE staining indicated a significant reduction in the cross-sectional area of gastrocnemius muscle fibers，with notable inflammatory cell infiltration and muscle atrophy in the model group. Masson's staining revealed obvious collagen fiber proliferation and deposition，with significant muscle fibrosis in the model group. The protein and mRNA expression levels of ATG5，Beclin1，LC3B，MuRF1，and MaFbx were significantly increased （P<0.01），while the protein expression of MYHC and IGF1 was significantly downregulated （P<0.01）. Compared with the model group，the high-dose group of Huangqi Guizhi Wuwutang showed significantly reduced protein and mRNA expression levels of ATG5，Beclin1，LC3B，MuRF1，and MaFbx （P<0.01） and increased protein expression levels of MYHC and IGF1 （P<0.01）. The cross-sectional area of muscle fibers increased，and the muscle cell morphology approached normal. Moreover，pathological abnormalities in the gastrocnemius muscle were significantly improved，with reduced collagen fiber proliferation （P<0.01）. ConclusionHuangqi Guizhi Wuwutang can mediate autophagy by regulating the expression of ATG5，Beclin1，LC3B，and IGF1，thereby reducing skeletal muscle catabolism and improving skeletal muscle homeostasis，which contributes to the treatment of RA-associated sarcopenia. The findings provide insight into the mechanisms underlying the effects of Huangqi Guizhi Wuwutang in the treatment of RA-related sarcopenia and offer a reference for its enhanced clinical application.

[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34⁺ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results.

ObjectiveTo investigate the effect and mechanism of polysaccharide in water extract of Cyclocarya paliurus （CPWP） in inhibiting benign prostatic hyperplasia （BPH）. MethodsCPWP was obtained by heating reflux， aqueous extraction， alcohol precipitation， and freeze drying. The chemical composition and structural properties of CPWP were analyzed by high performance liquid chromatography with 1-pheny-3-methyl-5-pyrazolone pre-column derivatization and infrared spectroscopy. Male SD rats were randomly assigned into control， model， finasteride （ig 5 mg·kg^-1）， and low-， medium-， and high-dose （ig 50， 75， 100 mg·kg^-1） CPWP groups， with 8 rats in each group. The BPH model was established by subcutaneously injecting propionate testosterone in castrated rats. The rats in the drug intervention groups were administrated with corresponding drugs， and those in the control group were administrated with an equal volume of normal saline each day. After 30 consecutive days， the rats were sacrificed， and the prostate tissue was separated and weighed. The effects of drug interventions on the body weight， prostate wet weight， and prostate index of rats were examined. The prostate tissue was stained with hematoxylin-eosin （HE） for observation of pathological changes. Enzyme-linked immunosorbent assay was employed to measure the level of dihydrotestosterone （DHT）， and immunohistochemical staining was used to detect the expression of steroid 5 alpha-reductase 2 （SRD5A2） and Ki67 in the prostate tissue. ResultsCPWP was identified as a saccharide， with characteristic absorption peaks of saccharides. CPWP showed the total sugar content of 44.15% and molecular weight within the range of 5.5-78.8 kDa， being composed of mannose， rhamnose， galacturonic acid， glucose， galactose， xylose， and arabinose. Compared with the control group， the model group had significantly increased prostate wet weight and prostate index （P<0.01）， thick and tall prostate epithelial cells， increased internal wrinkles， papillary expansion into the cavity， an elevation in DHT level in the serum， and up-regulated expression of SRD5A2 and Ki67 in the prostate tissue （P<0.05， P<0.01）. Compared with the model group， both the finasteride and CPWP groups showed decreases in prostate wet weight and prostate index （P<0.05， P<0.01）， thinned prostate epithelial cells， with only a small portion of internal wrinkles and papillary expansion into the cavity， shortened papillary protrusions， lowered DHT level in the serum， and down-regulated expression of SRD5A2 and Ki67 in the prostate tissue （P<0.01）. Moreover， CPWP exerted effects in a dose-dependent manner. ConclusionCPWP inhibits BPH by regulating the expression of SRD5A2.

Objective To explore the clinical efficacy of ultra-micro 8.0 mm single-port thoracoscopic nerve resection through areola incision in the treatment of primary palmar hyperhidrosis(PPH).Methods 84 patients with PPH from January 2018 to January 2022,were divided into the observed group[(inframammary approach,IMA)group,n=42]and control group[axillary approach(AA)group,n=42].The observed group was treated with ultra-micro 8.0 mm IMA single-port endoscopic thoracic sympathectomy(ETS),the control group used the traditional AA single-port ETS.The visual analogue scale(VAS)score,serum C-reactive protein(CRP),cortisol(Cor),interleukin-6(IL-6)levels,the postoperative cosmetic effect and compensatory hyperhidrosis of the two groups were compared.Results The VAS scores of 2,12 and 24 h after surgery in the observation group were significantly lower than those of the control group,the differences were statistically significant(P<0.05);At 12 and 24 h postoperation,the levels of CPR,Cor,and IL-6 levels in both groups of patients were significantly higher than preoperation,but those in the observation group were significantly lower than those of the control group,the differences were statistically significant(P<0.05);The total satisfaction rate of postoperative incision in the observation group was significantly higher than that of the control group,the difference was statistically significant(P<0.05);There was no statistically significant difference of the incidence of compensatory hyperhidrosis between the two groups(P>0.05).Conclusion The clinical efficacy of ultra-micro 8.0 mm IMA single-port ETS through areola incision in the treatment of PPH is good.Compared with the traditional axillary single-port thoracoscopic method,it has the advantages of small trauma,less bleeding,less patient pain,high safety and high patient satisfaction.It is worthy of clinical promotion and application.

Objectives:To observe the effectiveness and safety of recombinant human brain natriuretic peptide(rhBNP)for the treatment of patients with right heart failure caused by pulmonary arterial hypertension(PAH).Methods:A total of 421 patients with right heart failure caused by PAH who were hospitalized in Fuwai Hospital from January 2019 to June 2024 were retrospectively included in this study.All patients were treated with rhBNP on top of conventional therapy.24 h urine volume,body weight,liver and renal function index,electrolyte,uric acid,red blood cell distribution width(RBCDW),cardiac function,blood pressure and heart rate before and after the treatment of rhBNP were compared.Clinical symptoms,signs and the occurrence of adverse events during treatment were also observed.Results:Compared with baseline,after treatment with rhBNP,the 24 h urine volume increased.The levels of body weight,transaminase,total bilirubin,direct bilirubin,uric acid,RBCDW,systolic blood pressure,heart rate,and N-terminal pro-B-type natriuretic peptide significantly decreased(all P<0.05).There were no statistically significant differences in the levels of serum creatinine,and tricuspid annular plane systolic excursion to pulmonary artery systolic pressure ratio(both P>0.05).Dyspnea and lower limbs edema were improved in 75.2%cases(227/302)and 66.9%cases(281/420)respectively.The incidence of adverse events and severe adverse events during rhBNP treatment were 1.2%(5/421)and 0.5%(2/421)respectively.Conclusions:Adding rhBNP on top of standard medication can effectively increase 24 h urine volume,reduce body weight,improve some prognostic indicators,improve the clinical symptoms and signs of heart failure without negatively affecting the renal function in right heart failure patients caused by PAH.Blood pressure should be closely monitored during the treatment process.

The complement component 4 binding protein alpha(C4BPA)gene is the alpha chain of complement binding protein 4.As a plasma protein involved in the complement and coagulation systems,it can influence immune responses and lipid metabolism.In order to study the polymor-phism of C4BPA gene and its correlation with milk quality traits in Chinese Holstein cows,genom-ic DNA was extracted from blood samples of 92 Chinese Holstein cows,and the target fragment of C4BPA gene was amplified by PCR,and the association analysis was performed by using direct se-quencing to obtain the SNP loci and milk quality traits.The results showed that among the four SNPs found at the third intron of the C4BPA gene,I3-11 G＞A was highly significantly correlated with milk protein and urea nitrogen(P＜0.05),I3-291 T＞G was significantly correlated with lac-tose(P＜0.05),I3-374 C＞T was highly significantly correlated with lactose and urea nitrogen(P＜0.05),and I3-375 T＞G was highly significantly correlated with lactose(P＜0.05),milk pro-tein and urea nitrogen.The chi-square test values for each point indicated that the population was in genetic equilibrium.Individuals of haplotype combination H1 H1 had the highest lactose content,and haplotype combination H1H2 can be used as the best haplotype combination in the molecular selection work of dairy cows.

Aim To explore the predictive value of serum free triiodothyronine(FT3)on the long-term prognosis of patients with coronary heart disease after percutaneous coronary intervention(PCI).Methods All the subjects were from a prospective cohort study(PRACTICE study).In this study,15 250 patients with coronary heart disease after PCI in the First Affiliated Hospital of Xinjiang Medical University were selected,and the clinical data,FT3 and creatinine were collected.All the subjects were followed up regularly,and the primary follow-up endpoints were all-cause mortality and cardiogenic mortality,the secondary endpoints were major adverse cardiovascular events(MACE)and major adverse cardiovascular and cerebrovascular events(MACCE).According to the admission criteria,3 109 patients were finally in-cluded in this study.According to the baseline value of FT3,patients were divided into normal FT3 group(FT3:3.65～6.8 pmol/L,1 446 cases)and low FT3 group(FT3＜3.65 pmol/L,1 663 cases).Kaplan-Meier analysis was used for survival analysis,and Log-rank test was used for survival comparison.Multivariate Cox regression analysis was used to e-valuate the risk factors of the follow-up results of the two groups.Results Compared with the normal FT3 group,all-cause mortality and cardiogenic mortality in the low FT3 group increased significantly(P＜0.05).Kaplan-Meier analysis showed that the cumulative risk of all-cause mortality and cardiogenic mortality increased in the low FT3 group(P＜0.05).Multivariate Cox regression analysis indicated that the risk of all-cause mortality increased by 1.639 folds in the low FT3 group(HR=2.639,95％CI:1.385～5.348,P=0.007),while no statistical difference was found in cardiogenic mortality after adjusting for multiple factors(P=0.125).Conclusion The decrease in serum FT3 levels has important predictive value for all-cause mortality after PCI in patients with coronary heart disease.

Aim To investigate the role and mechanism of KLHL21 gene in myocardial infarction(MI)of mice.Methods KLHL21 gene knockout(KO)mice were generated using CRISPR/Cas9 technology,and C57BL/6 wild-type mice were used as controls.Sixty KLHL21 KO mice and 60 wild-type mice were randomly divided into four groups:WT+Sham group(n=30),WT+MI group(n=30),KO+Sham group(n=30)and KO+MI group(n=30).Postoperative is-chemic and infarct areas were assessed using TTC and Evans Blue staining,myocardial injury markers were measured by ELISA,cardiac function was evaluated by ultrasound,and histological changes were examined using HE and Masson stai-ning.Western blot was used to detect proteins related to the nuclear factor-κB(NF-κB)signaling pathway.Results KLHL21 protein expression in the myocardial tissue of KO mice was significantly lower than that in WT mice.The infarct area in KO+MI mice was significantly larger than that in WT+MI group.KO+MI mice showed reduced cardiac function compared with WT+MI mice.HE staining revealed myocardial cell loss,liquefactive necrosis,nuclear fragmentation,and significant neutrophil infiltration,while Masson staining showed aggravated fibrosis in KO+MI group.Serum tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),creatine kinase-MB(CK-MB),and cardiac troponin Ⅰ(cTnⅠ)lev-els were significantly increased in KO+MI mice compared with WT+MI mice.Western blot analysis showed increased lev-els of phosphorylated inhibitor of nuclear factor-κB alpha(p-IKBα),P65,and P50,and decreased nuclear factor-κB al-pha(IKBα)in KO+MI mice.Conclusion KLHL21 gene plays a preventive role in myocardial infarction in mice,possibly through inhibition of NF-κB signaling pathway activation.