Based on the traditional Chinese medicine （TCM） theories of body fluid metabolism and turbid phlegm obstructing the orifices， and drawing on recent advances in cerebrospinal fluid-interstitial fluid （CSF-ISF） exchange， glymphatic clearance， and extracellular matrix （ECM） regulation， this article discusses the possible scientific connotation of "invisible phlegm" from the perspectives of pathogenesis， clinical manifestations， and therapeutic implications. Impaired brain fluid exchange may lead to reduced clearance， the retention of metabolic wastes and pathological proteins. Abnormal ECM remodeling may further disrupt molecular diffusion and local microenvironmental homeostasis， resulting in a pathological process characterized by slowed fluid movement， reduced clearance， and abnormal retention. This process bears a certain functional resemblance to the TCM pathogenesis of impaired distribution of body fluids， endogenous formation of turbid phlegm and obstruction of the brain orifices， and may also be associated with manifestations such as a clouded sensation in the head， cognitive abnormalities， and emotional disturbances. On this basis， the potential effects of certain phlegm-dissolving Chinese medicines in improving brain clearance function and extracellular microenvironmental homeostasis warrant further investigation. We therefore propose that， the "invisible phlegm" of TCM in brain tissue may manifest as a pathological retention state characterized by restricted fluid exchange， impaired clearance function， and extracellular microenvironmental imbalance. This perspective may provide a new avenue for investigating TCM treatment of neurological and psychiatric disorders from the perspective of phlegm.

Based on the traditional Chinese medicine （TCM） theories of body fluid metabolism and turbid phlegm obstructing the orifices， and drawing on recent advances in cerebrospinal fluid-interstitial fluid （CSF-ISF） exchange， glymphatic clearance， and extracellular matrix （ECM） regulation， this article discusses the possible scientific connotation of "invisible phlegm" from the perspectives of pathogenesis， clinical manifestations， and therapeutic implications. Impaired brain fluid exchange may lead to reduced clearance， the retention of metabolic wastes and pathological proteins. Abnormal ECM remodeling may further disrupt molecular diffusion and local microenvironmental homeostasis， resulting in a pathological process characterized by slowed fluid movement， reduced clearance， and abnormal retention. This process bears a certain functional resemblance to the TCM pathogenesis of impaired distribution of body fluids， endogenous formation of turbid phlegm and obstruction of the brain orifices， and may also be associated with manifestations such as a clouded sensation in the head， cognitive abnormalities， and emotional disturbances. On this basis， the potential effects of certain phlegm-dissolving Chinese medicines in improving brain clearance function and extracellular microenvironmental homeostasis warrant further investigation. We therefore propose that， the "invisible phlegm" of TCM in brain tissue may manifest as a pathological retention state characterized by restricted fluid exchange， impaired clearance function， and extracellular microenvironmental imbalance. This perspective may provide a new avenue for investigating TCM treatment of neurological and psychiatric disorders from the perspective of phlegm.

Hashimoto thyroiditis(HT)is one of the most common autoimmune diseases."Wood depression and earth stagnation"is its important pathogenesis.Liver depression and qi stagnation,wood depression multiplies the earth,spleen deficiency and earth stagnation,phlegm and blood stasis intermingle,resulting in wood depression and earth stagnation.Or middle-earth stagnation insults wood in turn,liver yang deficiency,wood depression,and failure to disperse and discharge,resulting in earth stagnation and wood depression.It may be related to the intestinal immune mechanism in modern medical study.Based on the theory of"wood depression and earth stagnation",this article discussed the pathogenesis of HT from the perspective of wood depression and imbalance of microbe-gut-brain axis,earth stagnation and dysbiosis of intestinal flora,damage to the intestinal barrier,and proposed the treatment principles,i.e.,"disperse stagnated liver qi for relieving qi stagnation,ascend yang and regulate qi""eliminate stagnation and remove turbidity,invigorate spleen and restore normal movement",which could provide the ideas for mechanism research and clinical treatment of HT.

Objective To investigate the predictive value of triglyceride-glucose(TyG)for the risk of death in patients with meta-bolic dysfunction-associated steatotic liver disease(MASLD).Methods Examination data of adult MASLD patients were obtained from the National Health and Nutrition Examination Survey.A weighted multivariate COX proportional risk model was used to estimate the asso-ciation between TyG index and all-cause mortality and cardiovascular mortality.Segmented COX regression analysis was used to explore nonlinear relationships and associations between TyG index and all-cause mortality and cardiovascular mortality on either side of the in-flection point.Results Hazard ratios for all-cause mortality in the lowest to the highest quartiles of the TyG index were 1.00,1.01,1.17 and 1.36,respectively.For cardiovascular mortality,hazard ratios were 1.00,0.89,1.27 and 1.83,respectively.There was a U-shape relationship between the TyG index and all-cause mortality and cardiovascular mortality in patients with MASLD.Above the inflection point,each one-unit increase in TyG index was associated with a 25％increase in the risk of all-cause mortality and a 46％increase in the risk of cardiovascular mortality.Conclusion TyG index is U-shapedly associated with all-cause mortality and cardio-vascular mortality in patients with MASLD.Monitoring the TyG index is useful in assessing the risk of death in patients with MASLD.

Hashimoto thyroiditis(HT)is one of the most common autoimmune diseases."Wood depression and earth stagnation"is its important pathogenesis.Liver depression and qi stagnation,wood depression multiplies the earth,spleen deficiency and earth stagnation,phlegm and blood stasis intermingle,resulting in wood depression and earth stagnation.Or middle-earth stagnation insults wood in turn,liver yang deficiency,wood depression,and failure to disperse and discharge,resulting in earth stagnation and wood depression.It may be related to the intestinal immune mechanism in modern medical study.Based on the theory of"wood depression and earth stagnation",this article discussed the pathogenesis of HT from the perspective of wood depression and imbalance of microbe-gut-brain axis,earth stagnation and dysbiosis of intestinal flora,damage to the intestinal barrier,and proposed the treatment principles,i.e.,"disperse stagnated liver qi for relieving qi stagnation,ascend yang and regulate qi""eliminate stagnation and remove turbidity,invigorate spleen and restore normal movement",which could provide the ideas for mechanism research and clinical treatment of HT.

Objective To investigate the predictive value of triglyceride-glucose(TyG)for the risk of death in patients with meta-bolic dysfunction-associated steatotic liver disease(MASLD).Methods Examination data of adult MASLD patients were obtained from the National Health and Nutrition Examination Survey.A weighted multivariate COX proportional risk model was used to estimate the asso-ciation between TyG index and all-cause mortality and cardiovascular mortality.Segmented COX regression analysis was used to explore nonlinear relationships and associations between TyG index and all-cause mortality and cardiovascular mortality on either side of the in-flection point.Results Hazard ratios for all-cause mortality in the lowest to the highest quartiles of the TyG index were 1.00,1.01,1.17 and 1.36,respectively.For cardiovascular mortality,hazard ratios were 1.00,0.89,1.27 and 1.83,respectively.There was a U-shape relationship between the TyG index and all-cause mortality and cardiovascular mortality in patients with MASLD.Above the inflection point,each one-unit increase in TyG index was associated with a 25％increase in the risk of all-cause mortality and a 46％increase in the risk of cardiovascular mortality.Conclusion TyG index is U-shapedly associated with all-cause mortality and cardio-vascular mortality in patients with MASLD.Monitoring the TyG index is useful in assessing the risk of death in patients with MASLD.

Atherosclerosis is a vascular disease characterized by arterial occlusion formed by the pathological accumulation of pathological vascular cells and apoptotic cell debris. Atherosclerotic vulnerable plaque is an important pathological basis for inducing severe thrombotic cardiovascular events， and the study of its etiology and pathogenesis has always been a hot issue in the field of cardiovascular research. Efferocytosis is a new type of programmed death cell removal， which refers to the process of macrophages phagocytosing and degrading apoptotic cells to prevent secondary necrosis. It is a key homeostatic mechanism in the body's physiological process. In the pathological state， the dysfunction of efferocytosis causes the pathological accumulation of apoptotic cells and necrotic debris， leading to the occurrence of secondary cell necrosis and the continuous release of intracellular toxic content and inducing inflammatory regression disorders and cholesterol metabolism disorders， which are closely related to the occurrence and development of atherosclerotic vulnerable plaques. The theory of ''blood stasis and toxin'' is an important theory of traditional Chinese medicine （TCM） to explain the occurrence and development of atherosclerosis. Atherosclerosis starts from the pathological state of blood stagnation. Prolonged blood stagnation leads to blood stasis and toxic substances. Blood stasis and toxic pathogens interact with each other in blood vessels and eventually form plaques in blood vessels. The theory of ''blood stasis and toxin causing a catastrophe'' is an important understanding of the occurrence and development of acute cardiovascular events. From the perspective of TCM theory， the pathophysiological mechanism of efferocytosis is similar to the etiology and pathogenesis of the ''blood stasis and toxin'' in TCM. Therefore， this paper took the theory of ''blood stasis and toxin'' as the breakthrough point to explore the mechanism of efferocytosis in atherosclerotic vulnerable plaques， and proposed a detoxification and blood circulation method to regulate cell burial to prevent and treat atherosclerotic vulnerable plaques. The research strategy aims to provide new ideas and theoretical basis for the prevention and treatment of atherosclerosis by detoxification and blood circulation.

Objective:To investigate the protective effect of Epigallocatechin-3-gallate (EGCG) on triptolide (TP)-induced immunological liver injuries, and explore the relevant mechanisms of action. Methods: A total of 40 female C57BL/6 mice were randomly divided into four groups:control group,EGCG group,TP group and TP+EGCG group. The ALT levels in serum was examined by Reitman Frankel method. The activity of hepatic MDA,SOD and GSH was examined by spectrophotometry. HE staining was used to observed the changes of the hepatic histopathology. The hepatic IL-17,IL-6 levels was examined by ELISA and the expression of hepatic TLR4 protein was examined by Western blot. Results:The results showed that serum alanine aminotransaminase ( ALT) levels of TP group were obviously elevated (P<0. 005,vs normal control group) and serum ALT levels were obviously reduced in EGCG treatment group(P<0. 005,vs normal TP group). There were no significantly differences between EGCG group and control group (P>0. 05). Meanwhile,EGCG could ameliorate hepatic pathological damage. Furthermore,in TP group,the activity of malondialdehyde ( MDA) ,the expression of Toll-like receptor 4 (TLR4) protein and the contentration of hepatic interleukin (IL)-17,IL-6 were higher than normal control group ( P<0. 005 ) . On the contrary, the activity of superoxide dismutase ( SOD ) and restored glutathione ( GSH ) were significantly lower than normal control group ( SOD, P<0. 05;GSH, P<0. 005 ) . In EGCG treatment group, the expression of TLR4 protein and the concentration of MDA,hepatic IL-17 and IL-6 were lower than TP group ( TLR4,P<0. 05;MDA,P<0. 005;IL-17,P<0. 005;IL-6,P<0. 005). On the contrary,SOD and GSH were significantly higher than TP group (SOD,P<0. 05;GSH,P<0. 005). Conclusion:This study suggests that EGCG possesses hepatoprotective effect against TP-induced immunological liver injury through its anti-inflammatory and anti-oxidant actions.

Objective:To construct the Escherichia coli (E. coli)expression system for preparation of the bone morphogenetic protein-2 (BMP2)with collagen-binding domain (CBD),and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods:CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of recombinant protein was induced using isopropylβ-D-thiogalactopyranoside (IPTG) at 37 ℃.Ni-NTA chelate chromatography was used to purify CBD-BMP-2.Denaturing CBD-BMP2 was refolded by dilution method using ultrapure water.The refolding CBD-BMP2 was filtered through a 0.22μm microfiltration membrane for degermation.The recovery rate was calculated by the ratio of the protein concentration before and after degermation. The expression, purification, and renaturation of recombinant protein were detected by SDS-PAGE method.The concentration of CBD-BMP2 was detected by BCA assay.Results:The recombinant vector pet21b/CBD-BMP2 was successfully transformed into E.coli BL21,and the recombinant protein was expressed as inclusion bodies in E.coli.The SDS-PAGE results showed denaturing protein was dissolved in supernatant of lysis buffer with 8 mol·L-1 urea and the purified recombinant protein existed in elution buffer B with relative molecular mass about 14 000.Two bands (14 000 and 28 000)were seen in the SDS-PAGE picture,which indicated that the monomer was successfully refolded into dimer by dilution method.The concentrations of recombinant protein before and after degermation were 110 and 80 mg · L-1 , respectively, and the recovery rate was about 73%. Conclusion:The recombinant vector pet21b/CBD-BMP2 is transformed into E.coli BL21 successfully,and the recombinant CBD-BMP2 is expressed and refolded efficiently. The methods of prokaryotic expression system for preparing recombinant CBD-BMP2 protein are established.

Objective To observe the protective effect of gradual ischemic postconditioning (IP) capable of improving reperfusion on reperfusion injury in patients with ST-segment elevation acute myocardial infarction (STEAMl) undergoing primary percutaneous coronary intervention (PPCI).Methods 102 in-patients with STEAMI undergoing PPCI in the Department of Cardiology in the First People's Hospital of Kunshan City Affiliated to Jiangsu University from February 2011 to August 2014 were enrolled in this study. They were divided into three groups by a random number table: IP group (32 cases), gradual IP group (30 cases) and routine reperfusion group (40 cases). In IP group, after the opening of the infarction related blood vessel, ischemic postconditioning within the first minute of arterial reperfusion was made through three episodes of 1 minute inflation and 1 minute pressure withdrawn of an angioplasty balloon, and then persistent reperfusion was carried out. In the gradual IP group, the patients received three times of gradual angioplasty balloon inflation and denation, 1 minute/1minute, 30 seconds/30 seconds and 15 seconds/15 seconds respectively, presenting the gradual change of IP time. In the routine reperfusion group, after the opening of blocked blood vessel, the patients underwent routine PCI to persistently recover the coronary artery blood supply. The changes of related lead ST segment regression (Sum-STR), incidence of reperfusion arrhythmia, corrected thrombolysis in myocardial infarction (TIMI) franle count (CTFC), peaks of MB isoenzyme of creatine kinase (CK-MB), left ventricular ejection fraction (LVEF) and frequency of adverse events in follow-up period were compared among the three groups.Results The baseline characteristics were comparable in three groups. The incidence of ventricular premature beats was significantly lower in gradual IP group than that in routine reperfusion group [30.0% (9/30) vs. 55.0% (22/40),P < 0.05], and although theincidence of ventricular premature beats was lower in IP group than that in routine reperfusion group [34.4% (11/32) vs. 55.0% (22/40)], no statistically significant difference was found (P > 0.05). The incidence of ventricular tachycardia was significantly lower in IP and gradual IP groups than that in routine reperfusion group [15.6% (5/32), 13.3% (4/30) vs. 40.0% (16/40), bothP < 0.05]. The incidences of ventricular fibrillation, bradyarrhythmia and sinus arrest were lower in IP group and gradual IP group than those in routine reperfusion group, but no statistically significant differences were found (allP > 0.05). In IP group and gradual IP group, the Sum-STR incidence, CTFC, CK-MB peaks were lower than those of routine reperfusion group [Sum-STR: (56.7±18.3)%, (57.3±21.5)% vs. (44.6±21.6)%; CTFC: 25.47±5.37, 24.46±6.41 vs. 31.62±7.56; CK-MB peaks (U/L): 126.3±78.5, 121.6±82.5 vs. 147.4±72.5; allP < 0.05], the effect of gradual IP group being the best among the three groups. The levels of LVEF were slightly higher in IP and gradual IP groups than the level in routine reperfusion group (0.507±0.042, 0.511±0.062 vs. 0.497±0.062), but no statistically significant difference was found (bothP > 0.05). In routine reperfusion group, one patient died because the ventricular fibrillation could not be corrected and another one died of no-reflow during operation. Each group had 1 patient died during the 4 weeks of follow-up after operation, in the routine reperfusion group, one died of refractory heart failure, and the cause of death of other two patients, one in IP group and another in gradual IP group, was considered due to subacute thrombosis in stent. Major bleeding events were not found in each group.Conclusion Gradual IP can ameliorate myocardial reperfusion injury more significantly in patients with STEAMI undergoing PPCI.