Traditional four examinations of traditional Chinese medicine （TCM） are based on the symptoms and signs of patients， which are the advantages of TCM but also have shortcomings. Chronic kidney disease has the characteristics of insidiousness， long-term， deficiency and variability during its onset， which are difficult to be intervened in time based on only symptoms， therefore it is necessary to extend the application of the four examinations in the diagnosis and treatment process of chronic kidney disease. Based on the background of the continuous development of TCM syndrome differentiation techniques， this article proposed the extension and application strategies of the traditional four examinations in chronic kidney disease， including the incorporation of microscopic syndrome differentiation to identify the causes of kidney disease and prevent symptom deterioration； the utilization of accurate examination information enhanced by artificial intelligence for controlling development of existing disease； the integration of disease differentiation and syndrome differentiation to summarize clinical rules towards using constant to measure variation； and the establishment of a kidney disease database for the storage of four examinations information to prevent recurrence after recovery. The four above extension and application strategies can be used to achieve the long-term management and treatment effects of timely and early diagnosis， dynamic observation of the condition， accurate application of intervention， and strengthened prognosis assessment in the diagnosis and treatment of chronic kidney disease， and expand the advantages of TCM in the prevention and treatment of chronic kidney disease.

ObjectiveTo investigate the effect of Fuzheng Huayu prescription on hepatocyte extinction and regeneration in fibrotic liver and its mechanism of action in promoting hepatocyte regeneration. MethodsMice were given intraperitoneal injection of CCl₄ for 6 weeks to establish a model of liver cirrhosis， and there were 10 mice in the model group， 10 in the sorafenib group， 10 in the Fuzheng Huayu prescription group， and 9 in the normal control group. Since week 4 of modeling， the mice in the Fuzheng Huayu prescription group and the sorafenib group were given the corresponding drug by gavage at a dose of 4.8 g/kg and 4 mg/kg， respectively， for three consecutive weeks， and those in the normal group and the model group were given an equal volume of sodium carboxymethyl cellulose. Serum liver function parameters were measured； the METAVIR scoring system was used to evaluate liver inflammation and fibrosis stage； Sirius Red staining and hydroxyproline （Hyp） content in liver tissue were used to evaluate collagen deposition； immunohistochemistry was used to measure the protein expression levels of type IV collagen， CD31， CD32b， Ki67， CyclinD1， glutamine synthetase， Wnt2， and HGF， and Western blot was used to measure the expression levels of Wnt2， LRP6， β-catenin， p-β-catenin， and CyclinD1 in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， the Fuzheng Huayu prescription group and the sorafenib group showed the following changes： significant reductions in the serum levels of alanine aminotransferase and aspartate aminotransferase and the content of Hyp in liver tissue （all P<0.01）； a significant reduction in METAVIR score； significant reductions in the expression levels of type Ⅳ collagen and CD31 （all P<0.05） and a significant increase in the expression level of CD32b （P<0.01）； significant reductions in the number of parenchymal extinction lesions and significant increases in the expression levels of Ki67 and CyclinD1 in liver tissue （all P<0.01）； significant increases in the protein expression levels of Wnt2， LRP6， β-catenin， and CyclinD1 and a significant reduction in the protein expression level of p-β-catenin （all P<0.05）； significant increases in the number of cells stained positive for both CD32b and Wnt2. ConclusionFuzheng Huayu prescription can inhibit hepatic sinusoidal capillarization， improve the Wnt2 exocrine function of liver sinusoidal endothelial cells， activate the Wnt/β-catenin signaling pathway associated with hepatocyte regeneration， and finally reverse liver cirrhosis.

Objective To explore the expression and significance of neutrophil extracellular traps(NETs)in peripheral blood of primary Sj?gren's syndrome(pSS)patients across different disease activity levels,and the predictive value of NETs and routine laboratory markers antithrombin Ⅲ(AT Ⅲ),alkaline phosphatase(ALP)and carbohydrate antigen 125(CA125)for pSS disease activity.Methods A total of 94 newly diagnosed pSS patients at the Affiliated Lianyungang Hospital of Xuzhou Medical University from October 2021 to December 2023 were categorized into active(n=49)and non-active(n=45)groups based on the European League Against Rheumatism(EULAR)Sj?gren's Syndrome disease activity index(ESSDAI).The levels of NETs biomarkers,namely serum myeloperoxidase(MPO)-DNA,were measured using ELISA.Laboratory routine indicators and MPO-DNA were integrated into multivariate Logistic regression to screen for independent influencing factors of pSS disease activity.Pearson's correlation was used to evaluate the relationship between MPO-DNA levels and ESSDAI scores.The efficacy of MPO-DNA alone or in combination with AT Ⅲ,ALP and CA125,for predictors of disease activity was evaluated using ROC curve.Results Serum MPO-DNA(23.884±3.494 μg/L),ALP(80.159±34.318 U/L)and CA125(20.300±16.560 U/ml)levels of active pSS patients were higher than those in the non-active patients(19.024±3.324 μ g/L,67.500±21.166U/L,13.200±6.340 U/ml),while AT Ⅲ(89.180±15.040 ng/ml)was lower than that in non-active patients(94.650±11.620 ng/ml),with significant differences(t=-7.921,-2.426,-2.925,2.094,all P＜0.05).Multivariate Logistic regression analysis showed laboratory indicator MPO-DNA,ALP and CA125 were independent risk factors,while AT Ⅲ was an independent protective factor(all P＜0.05).MPO-DNA was positively correlated with ESSDAI scores(r=0.602,P＜0.01).The AUC(95％CI)of the combination of ALP,CA125 and AT Ⅲ for predicting disease activity in pSS was 0.711(0.612～0.810).The AUC(95％CI)of MPO-DNA alone for predicting disease activity in pSS was 0.837(0.758～0.915),and the AUC(95％CI)of combination of MPO-DNA,ALP,CA125 and AT Ⅲ for predicting disease activity in pSS was 0.866(0.797～0.935),showing an improving in the predictive value.Conclusion The involvement of NETs in the occurrence and expression levels of pSS is related to its disease activity.NETs combined with ALP,CA125 and AT Ⅲ have effective diagnostic performance for the disease activity of pSS.This tool can serve as a biological indicator for predicting the disease activity of pSS.

[Objective]To summarize the experience of YE Tianshi in the treatment of kidney diseases by unblocking method,so as to provide ideas for clinical differentiation and treatment of kidney diseases.[Methods]Taking Clinical Practice with Medical Records as the research object,combining with the discussion of ancient books such as Inner Canon of Yellow Emperor and Treatise on Febrile and Miscellaneous Diseases,the experience of YE Tianshi's treatment of kidney diseases with unblocking method was summarized.[Results]Unblocking method has a rich connotation.All methods that help the flow of Qi and blood and restoring the normal physiological state of the organism can be considered as unblocking method.YE Tianshi put forward the academic proposition of"all diseases should be treated with unblocking method",and established many methods such as dredging collaterals,promoting free flow of Qi,removing blood stasis and promoting diuresis,activating Yang,which inspired the clinical application of unblocking method.In the identification and treatment of kidney diseases,YE Tianshi attached great importance to the pathogenesis of obstruction of kidney Qi,and considered that the disharmony of Yin and Yang,the pathogen of phlegm,dampness and blood stasis were accumulated,eight extraordinary meridians were blocked,deficiency interweaved with excess was the key to cause kidney diseases,treatment should pay attention to moving kidney Qi.In the treatment of edema,stranguria with turbid discharge disease,spermatorrhea and other kidney diseases,formed the treatment principles of"regulating Yin and Yang,functioning normally when unobstructed""removing blood stasis and dredging collaterals,opening the orifices and invigorating the kidney""dredging the eight extra meridians,tonifying the liver and kidney",selected prescription and dispatching drugs,and achieved remarkable effects.[Conclusion]YE Tianshi's application of unblocking method in the treatment of kidney diseases has its own characteristics,which is accurate in syndrome differentiation,clear in theory and simple in medication,and provides a way of thinking for the application of unblocking method in the treatment of kidney diseases.

Objective:To measure the overall and lobulated volume of the liver with different degrees of liver fibrosis and to further observe pathological changes such as liver microvasculature, hepatocyte apoptosis, and regeneration in order to understand the macroscopic volume changes of the liver during liver fibrosis and its relationship with liver tissue microscopic pathology in patients with chronic liver disease.Methods:53 patients with chronic hepatitis B, alcoholic fatty liver disease, autoimmune liver disease, nonalcoholic fatty liver disease, and drug-induced chronic liver disease who underwent both liver biopsy tissue and abdominal magnetic resonance imaging were collected. Patients were divided into early (F1-2), middle (F3-4), and late (F5-6) in accordance with the Ishak fibrosis stage and Masson stain. The liver and spleen volumes were measured using ITK-SNAP software. CD31 immunohistochemical staining was used to reflect intrahepatic angiogenesis. Ki67 and HNF-4α multiplex immunohistochemical staining were used to reflect hepatocyte regeneration. GS staining was used to determine parenchymal extinction lesions. TUNEL staining was used to observe hepatocyte apoptosis. Spearman correlation analysis was used to analyze the relationship between liver volume changes and liver histopathological changes.Results:As liver fibrosis progressed, the total liver volume and right lobe liver volume gradually decreased ( P<0.05), while the spleen volume gradually increased ( P<0.05). The expression of CD31 and GS gradually increased ( P<0.05), and the expression of Ki67 first increased and then decreased ( P<0.05). The positivity rate of CD31 was negatively correlated with the right lobe liver volume ( r=-0.609, P<0.001) and the total liver volume ( r=-0.363, P=0.017). The positivity rate of Ki67 was positively correlated with the right lobe liver volume ( r=0.423, P=0.018), while the positivity rate of apoptotic cells was significantly negatively correlated with the total liver volume ( r=-0.860, P<0.001). The positivity rate of GS was negatively correlated with the right lobe liver volume ( r=-0.440, P=0.002), and the number of PELs was negatively correlated with RV ( r=-0.476, P=0.013). The CD31 positive staining area was negatively correlated with the Ki67 positive staining area( r=-0.511, P=0.009). Conclusion:As liver fibrosis progresses, patients with chronic liver disease have a depletion in total liver volume and right lobe liver volume, and this is mainly in correlation with fewer liver cells and liver tissue microvasculature disorders.

Chloroquine has played a pivotal role in the treatment of malaria.Recently,with the development of clinical application,they are increasingly recognized for their effectiveness in other as-pects,such as anti-inflammation,immunosuppression,anti-amoebic effects and so on.In the outbreak of the new coronavirus infection,chloroquine and its derivative hydroxychloroquine swiftly included in the treatment of infected patients in many countries.However,several studies have shown these drugs have some detrimental influence,especially in cardiac arrhythmia(prolonged the QT interval and in-creased risk of torsional ventricular tachycardia).Thus,we should pay attention to both their benefi-cial effects and potential adverse manifestation.In order to regulate the use of drugs and guide the clin-ic,this paper reviewed the electrophysiological basis and clinical characteristics of cardiac adverse re-actions of these drugs from the perspective of ion channels.

Chloroquine has played a pivotal role in the treatment of malaria.Recently,with the development of clinical application,they are increasingly recognized for their effectiveness in other as-pects,such as anti-inflammation,immunosuppression,anti-amoebic effects and so on.In the outbreak of the new coronavirus infection,chloroquine and its derivative hydroxychloroquine swiftly included in the treatment of infected patients in many countries.However,several studies have shown these drugs have some detrimental influence,especially in cardiac arrhythmia(prolonged the QT interval and in-creased risk of torsional ventricular tachycardia).Thus,we should pay attention to both their benefi-cial effects and potential adverse manifestation.In order to regulate the use of drugs and guide the clin-ic,this paper reviewed the electrophysiological basis and clinical characteristics of cardiac adverse re-actions of these drugs from the perspective of ion channels.

【Objective】 To investigate the effect of polymerized human cord hemoglobin (PolyCHb) on the chemosensitivity of human breast cancer MCF-7 cell subcutaneous xenografts in nude mice and its mechanism. 【Methods】 The MCF-7 cells in exponential growth phase were collected and made into suspension cells at a density of 5×10⁷ cells/mL.Subsequently, the cells were inoculated subcutaneously in the right limb of 18 BALB/c-nu nude mice with 0.2 mL cells per mouse to establish subcutaneous xenograft.When the tumor volume reached about 100 mm³, they were randomly divided into chemotherapy group: doxorubicin 5 mg·kg^-1, once/week; chemotherapy + PolyCHb group: in addition to doxorubicin (chemotherapy group), PolyCHb 600 mg·kg^-1, 3 times/week; the control group: normal saline 90 mg·kg^-1, once/week; all were injected through tail vein continuously for 4 weeks.From the day of injection (d 0), the tumor volume of each group of nude mice was measured every 3 days, and the tumor growth curves were drawn accordingly.After 38 days, the tumor growth observation was completed.The tumor was removed and weighed to calculate the tumor inhibition rate.HE staining, immunohistochemistry and TUNEL method were used to observe the pathological changes of tumor tissue, detect the expression of HIF-1α, and detect tumor cell apoptosis respectively.The content of reactive oxygen species (ROS) of each group was determined by fluorescence staining. 【Results】 The tumor volume (mm³) of chemotherapy + PolyCHb group, chemotherapy group and the control group at day 38 were 196.35±103.45 vs 316.29±62.88 vs 519.42±177.33 (P<0.05), and the tumor inhibition rate (%) of chemotherapy + PolyCHb treatment group and chemotherapy group was 62.20 vs 39.11, respectively.HE staining and TUNEL detection showed that cell necrosis and apoptosis in the growth area of tumor tissue increased in chemotherapy + PolyCHb group.Immunohistochemistry and fluorescence staining showed that HIF-1α expression in chemotherapy + PolyCHb group decreased and reactive oxygen species (ROS) content increased. 【Conclusion】 PolyCHb increases the chemosensitivity of subcutaneous xenograft in nude mice with breast cancer, and its mechanism may be related to the increase of ROS in tumor tissue and the promotion of tumor cell apoptosis.

【Objective】 To investigate the application prospect of platelet-rich plasma lysate (PL) microneedles in the treatment of wound healing by optimizing the preparation process and evaluating the activity of platelet growth factor. 【Methods】 Firstly, hyaluronic acid (HA) was used as needle matrix material, and the concentration of HA was preliminarily determined by investigating the fluidity of HA solution, shape of needle, pressure change performance and puncture performance.Combined with the results of HA dissolution in PL, mechanical properties of PL microneedles and skin puncture, the preparation process of platelet-rich plasma lysis microneedles (PLMN) was optimized to investigate the growth factor activity of PLMN and evaluate the effect of PLMN on wound healing in rats. 【Results】 When the concentration of HA was 30 mg/mL, the solution had good fluidity.The HA microneedles could pierce three layers of sealing film, with the puncture rate at 70%, and the solution had good mechanical properties.When HA was dissolved in PL at a concentration of 30 mg/mL at 4℃, the solution had no crystallization and was with good fluidity.The prepared PLMN showed good needle shape and no bending under HD microscope and scanning electron microscope.In vitro transdermal simulation results showed that PLMN could penetrate 4 layers of sealing membrane, and the puncture rate of the first three layers was more than 85%.In vivo lysis experiment showed that PLMNs could be completely dissolved within 10 min after being inserted into the body.Forty minutes later, the micropores left after the insertion of the microneedle were almost closed, without redness, swelling, erythema and other adverse reactions.In vitro and in vivo transdermal experiments showed that clear blue holes were visible after PLMNs penetrated the skin stain, indicating that the microneedles could pierce the skin barrier.Growth factor detection results showed that the content of platelet-derived growth factor (PDGF-BB) and transforming growth factor (TGFβ1) in platelet lysate were (17.67±1.23) ng/mL and (105.95±2.16) ng/mL, respectively.After PLMNs were prepared and stored at room temperature for 1 week, the content of PDGF-BB(ng/mL) and TGFβ1(ng/mL) were 15.48±2.12 and 56.26±1.53, respectively.Compared with PL(vs 17.67±1.23 and 105.95±2.16), the content of PDGF-BB and TGFβ1 decreased, but still maintained the activity of growth factors.The results of wound healing experiment in rats showed that the wound healing rate (%) of PLMN group on the third day was significantly different from that of the control group(52.51±6.15 vs 33.31±3.60, P<0.05). 【Conclusion】 The PLMN prepared in this experiment has good mechanical properties and can successfully pierce the skin cuticle.It can also maintain the activity of growth factors, and has a good effect on wound healing in rats, which provides a new idea for minimally invasive treatment in the field of tissue repair.

【Objective】 To investigate the removal efficacy of inflammatory cytokines and blood compatibility of modified PBTNF. 【Methods】 Acrylic acid (AA) was firstly UV-grafted onto the surface of PBTNF to negatively charge the surface of the material. Subsequently, the three positively charged polyelectrolytes, DA, PEI, and CS were respectively electrostatic self-assembled with GO on the surface of PBTNF, forming two layers of film with GO as the outer layer: PBTNF-(DA/GO)2, PBTNF-(PEI/GO)2, PBTNF-(CS/GO)2. 【Results】 Scanning electron microscopy results showed that compared with the PBTNF grafted with AA, the adhesion of particles was observed on the surface of the three modified materials, and the photo shows that the color of the material surface was deepened after electrostatic self-assembly. The results of wettability showed that the surface hydrophilicity was significantly improved, indicating that the electrostatic self-assembled membrane was successfully immobilized on the surface of PBTNF. The removal efficiency (%) of IL-1β for PBTNF-(DA/GO)2, PBTNF-(PEI/GO)2 and PBTNF-(CS/GO)2 were 69.00±7.36 vs -2.35±2.69 vs -1.59±3.26 (P<0.05). The removal efficiency of IL-6 (%) were 40.15±1.86 vs -13.46±5.72 vs -1.21±3.41 (P<0.05). The removal efficiency of IL-8 (%) were 96.90±0.97 vs 17.84±11.74 vs 43.68±17.38 (P<0.05). The removal efficiency of TNF-α (%) was 44.46±2.50 vs 14.90±7.12 vs 20.64±1.22 (P<0.05). Plasma protein adsorption results (total protein, immunoglobulin G, albumin) and red blood cell deformability index showed that there was no statistical difference among the three modified PBTNFs and the control group (P>0.05). Although the red blood cell osmotic fragility (g/L) of the three modified PBTNFs is higher than that of the former: control group vs PBTNF-(DA/GO)2 vs PBTNF-(PEI/GO)2 vs PBTNF-(CS/GO)2: 4.39±0.05 vs 4.62±0.02 vs 4.48±0.03 vs 4.90±0.03 (P<0.05), the hemolysis rate (%) of them were all less than 5%, and PBTNF-(DA/GO)2 performed the lowest hemolysis rate which was (0.03±0.01)% (compared with PBTNF-(PEI/GO)2, P<0.05). The coagulation function test results showed that compared with the control group, the fibrinogen (g/L) of the three modified PBTNFs had no statistical difference (P>0.05); the activated partial thrombo plastin time (S) slightly extended, but all within the normal range of clinical standard; and the prothrombin time (S) of PBTNF-(CS/GO)2 was prolonged(P<0.05). 【Conclusion】 Among the three positively charged polyelectrolytes, including DA, PEI, and CS, PBTNF-(DA/GO)2 performed the best removal rate of inflammatory cytokines, and the blood compatibility evaluation results showed that PBTNF-(DA/GO)2 had no significant effect on red blood cells and coagulation function. Consequently, in the study of inflammatory cytokines adsorption, DA is expected to be the optimal polyelectrolyte assembling with GO for further research.