Diabetic retinopathy（DR）， as one of the most common and serious microvascular complications of diabetes mellitus， seriously threatens human health， and belongs to "Xiaoke eye diseases" in traditional Chinese medicine（TCM）， which has been richly experienced by medical practitioners through the ages， but is mostly recorded in a piecemeal manner and has not been systematically researched. This disease is featured by long course and repeated attack， and is refractory， which belongs to the research category of "persistent illness entering collaterals". Systematic establishment of TCM collateral disease theory for guiding prevention and treatment of DR has important clinical value. On the basis of close correlation between tertiary collaterals at the terminal of collaterals and capillaries and microcirculation， the concept of "tertiary collaterals-microvascular" is proposed. It is pointed out that DR falls within the scope of "tertiary collaterals-microvascular" diseases， and presents four types of micro-pathological characteristics， including stasis， insufficiency， growth and bleeding of tertiary collaterals. It is concluded that "deficiency of both Qi and Yin" is the basic pathogenesis of DR， and "blood stasis and collateral obstruction" is the important pathogenesis and key factor. Thus， the treatment method of "dispersing blood stasis， dredging collateral， tonifying Qi and Yin， stopping hemorrhage and improving eyesight" is determined， and the formula of Tongluo Mingmu capsules is developed. The article tightly focuses on the pathological changes such as stasis， growth， insufficiency and bleeding of collaterals， addresses both symptoms and root causes， and plays a synergistic role of both dispersing stasis and stopping bleeding. In this way， it can realize the purpose of tonifying Qi and Yin to replenish the essence， dispersing stasis and dredging collaterals to meet the requirement， as well as stopping hemorrhage and improving eyesight to deal with changes. Fundamental researches demonstrate that Tongluo Mingmu capsules has synergy effects of protecting both retinal capillaries and retinal cells. Phase-Ⅲ clinical trial of new drug has proven definite clinical efficacy and good safety， which provides a new drug choice for enhancing clinical effect of DR， and further supports the scientific value of Luobing theory in preventing and treating DR and other clinically significant diseases.

In the Qing Dynasty， YE Gui proposed the pathogenesis of "chronic diseases entering the collaterals" as a rule for collateral diseases. In the context of viral respiratory diseases， the course of the disease is considered a relative concept. A systematic review of historical literature reveals that "new pathogens" belong to the category of "new diseases". It identifies that "new diseases" include types where the pathogenic qi initially invades without harming the righteous qi， where old diseases relapse while not being completely cured， and where chronic diseases suddenly flare up. These are not limited to exogenous diseases. "New pathogens" refer to diseases caused by the invasion of warm-heat pathogens， the six excesses， and epidemic qi， differing from the modern traditional Chinese medicine term "new warm diseases" related to latent heat diseases. Based on the "collaterals-meridians-collaterals" transformation pathway described in The Inner Canon of Yellow Emperor （《黄帝内经》） and the pathogenesis of external pathogens entering the zang-fu organs via the meridians as described in Treatise on Febrile Disease （《伤寒论》）， as well as the external warm diseases' transformation rule explained by YE Gui from the perspective of collaterals， the concept of "new pathogen invading the collaterals" is proposed. Further analysis indicates that this concept aligns with the transmission and pathogenesis patterns of viral respiratory diseases. The treatment method of "clearing epidemic pathogens， detoxifying， and spreading the lung to relieve heat" is considered important for the prevention and control of viral respiratory infectious diseases.

ObjectiveExploring the formula rules of commonly used traditional Chinese medicines（TCMs） for epidemic treatment from the Qin and Han dynasties to the Qing dynasty through data mining， providing reference for the prevention and control of contemporary epidemics. MethodsThe articles on epidemic treatment in the electronic database of Chinese Medical Code V5.0 were systematically searched， and the contents such as source， dynasty， author， diagnosis， formula name， therapeutic method and efficacy， and composition of medicines from each article that met the inclusion criteria were extracted. Then， an Excel standardized database was established， and Python programs were used for data mining to summarize the frequency of commonly used medicines and perform hierarchical cluster analysis， Pearson correlation analysis， and association rule analysis. ResultsA total of 1 595 formulas were included， involving 558 TCMs. The efficacy of these medicines could be classified into two categories， namely， expeling pathogenic factors and reinforcing healthy Qi. According to the frequency deconstruction analysis， high-frequency medicines were mainly detoxification， Fu-organ dredging， aromatization and promoting blood circulation， followed by the medicines with the effect of treating the lungs， such as clearing the lungs and resolving phlegm， clearing heat and purging the lungs， relieving cough and asthma， and purging the lungs and relieving asthma. And the proportions of acrid-warm herbs and acrid-cold herbs varied in different periods. Hierarchical clustering and correlation analysis both suggested TCMs for expeling pathogenic factors and reinforcing healthy Qi often formed stable combinations with high association degrees. Association rule analysis showed that the core acrid-warm herb was mainly Ephedrae Herba， and the core acrid-cold herb was mainly Forsythiae Fructus， resulting in the core formulas of Maxing Shigantang and Yinqiaosan. ConclusionThroughout history， the prevention and control of epidemics have been based on the principle of "preserving healthy Qi and avoiding toxic Qi"， focusing on the treatment of the causes and characteristics of epidemics through detoxification， Fu-organ dredging， and aromatization， emphasizing the use of Rhei Radix et Rhizoma and other herbs to dredge Fu-organ， eliminate toxins and pathogens， and playing the role of actively intervene with symptomatic medication. And based on the external manifestations of the body's struggle between evil and righteousness， diagnose and treatment according to syndrome differentiation was performed.

ObjectiveExploring the formula rules of commonly used traditional Chinese medicines（TCMs） for epidemic treatment from the Qin and Han dynasties to the Qing dynasty through data mining， providing reference for the prevention and control of contemporary epidemics. MethodsThe articles on epidemic treatment in the electronic database of Chinese Medical Code V5.0 were systematically searched， and the contents such as source， dynasty， author， diagnosis， formula name， therapeutic method and efficacy， and composition of medicines from each article that met the inclusion criteria were extracted. Then， an Excel standardized database was established， and Python programs were used for data mining to summarize the frequency of commonly used medicines and perform hierarchical cluster analysis， Pearson correlation analysis， and association rule analysis. ResultsA total of 1 595 formulas were included， involving 558 TCMs. The efficacy of these medicines could be classified into two categories， namely， expeling pathogenic factors and reinforcing healthy Qi. According to the frequency deconstruction analysis， high-frequency medicines were mainly detoxification， Fu-organ dredging， aromatization and promoting blood circulation， followed by the medicines with the effect of treating the lungs， such as clearing the lungs and resolving phlegm， clearing heat and purging the lungs， relieving cough and asthma， and purging the lungs and relieving asthma. And the proportions of acrid-warm herbs and acrid-cold herbs varied in different periods. Hierarchical clustering and correlation analysis both suggested TCMs for expeling pathogenic factors and reinforcing healthy Qi often formed stable combinations with high association degrees. Association rule analysis showed that the core acrid-warm herb was mainly Ephedrae Herba， and the core acrid-cold herb was mainly Forsythiae Fructus， resulting in the core formulas of Maxing Shigantang and Yinqiaosan. ConclusionThroughout history， the prevention and control of epidemics have been based on the principle of "preserving healthy Qi and avoiding toxic Qi"， focusing on the treatment of the causes and characteristics of epidemics through detoxification， Fu-organ dredging， and aromatization， emphasizing the use of Rhei Radix et Rhizoma and other herbs to dredge Fu-organ， eliminate toxins and pathogens， and playing the role of actively intervene with symptomatic medication. And based on the external manifestations of the body's struggle between evil and righteousness， diagnose and treatment according to syndrome differentiation was performed.

As a public health event that seriously threatens human health， viral respiratory infectious diseases fall within the scope of "new illness entering collaterals". Collateral disease theory is used to guide prevention and treatment of this type of diseases， thus to propose the concept of infectious lung disease epidemic and reveal its progress rule of "Qi collaterals-Qi pathway-blood collaterals". By analyzing main therapeutic methods including detoxication， precipitation draining and aromatization， this paper proposes active intervention and treatment strategy of "simultaneously treating Wei and Qi， relieving exterior-interior syndrome， using medicine with evidence， stopping the disease tendency， overall adjustment and multi-target therapy". Through systematic summarization of historical medication regularity in infectious lung disease epidemic， this paper demonstrates that "maintaining healthy Qi" and "avoiding harmful Qi" are the general rules of prevention and treatment. Based on literature review， data mining and meta analysis of clinical experiences， this study inherits and innovatively develops the formula of Chinese patent medicine Lianhua Qingwen. The formula is developed through a modification to basic formulae Yinqiaosan and Maxing Shigantang. It learns from Wu Youke's experience in treating epidemic diseases with Rhei Radix et Rhizoma， considers pathological characteristics of infectious lung disease epidemic， and innovatively adds Rhodiolae Crenulatae Radix et Rhizoma and Pogostemonis Herba. The new formula is featured by clearing heat， being warm-pungent， function of both detoxication and aromatization， as well as simultaneous implementation of strengthening the body resistance and purging Fu organs. It can clear heat but won't over cool the body， keep the body warm but won't contribute to pathogenic fire， strengthening the body resistance but won't retain pathogen， and eliminate pathogen but won't damage healthy Qi. Meanwhile， series of basic and clinical studies have demonstrated that Lianhua Qingwen formula has the unique advantages for respiratory infectious diseases， which reveals the scientific connotation of infectious lung disease prevention and control under the guidance of pulmonary collateral disease diagnosis and treatment theory.

ObjectiveTo investigate the effect and mechanism of ginsenoside Rg₁ （G-Rg₁） combined with hirudin in treating myocardial fibrosis in the mouse model of acute myocardial infarction （AMI）. MethodSeventy-five C57BL/6N mice were randomized into sham， model， G-Rg₁ （20 mg·kg^-1）， hirudin （20 mg·kg^-1）， and G-Rg₁ （20 mg·kg^-1） + hirudin （20 mg·kg^-1） groups. The mouse model of AMI was established by ligation of the anterior descending branch of the left coronary artery and continued gavage for 4 weeks. The success of the modeling was judged by ECG changes of mice before and after ligation. The heart weight index， echocardiography， myocardial fibrosis， type Ⅰ collagen α1（COL1A1）， platelet-endothelial cell adhesion molecule-1 （PECAM-1/CD31）， α-smooth muscle actin （α-SMA）， intercellular adhesion molecule-1 （ICAM-1）， and vascular cell adhesion molecule-1 （VCAM-1） were measured before and after treatment. ResultAfter ligation of the anterior descending branch of the left coronary artery， the R wave and T wave merged into a tall tented wave， and the ST segment presented a "damaged" change， indicating that the model was successfully prepared. Compared with the sham group， the model group showed dull and dry hair， slow movement， increased heart weight index （P<0.01）， decreased left ventricular ejection fraction （EF） and left ventricular shortening fraction （FS） （P<0.01）， increased left ventricular end-diastolic diameter （LVEDd） and left ventricular end-systolic diameter （LVESd） （P<0.01）， disarranged myocardial fibers， collagen fiber hyperplasia （P<0.01）， increased expression of COL1A1 （P<0.01）， upregulated protein levels of ICAM-1 and VCAM-1 （P<0.01）， downregulated CD31 expression， and upregulated α-SMA expression （P<0.01）. Compared with the model group， the treatment groups recovered the above indexes in different degrees （P<0.05， P<0.01）， and the combined group had better effect （P<0.05）. ConclusionG-Rg₁ combined with hirudin can ameliorate myocardial fibrosis after AMI by inhibiting the expression of adhesion molecular protein in the heart tissue， reducing the adhesion of inflammatory cells， alleviating cardiac inflammation， and inhibiting cardiac endothelial-to-mesenchymal transition.

AIM:To investigate the effects of Balanophora involucrata Hook.f.(BIH)extracts on bile acid metabolism and liver injury in mice with metabolic dysfunction-associated fatty liver disease(MAFLD)through the gut mi-crobiota-farnesoid X receptor(FXR)axis,and to explore the underlying mechanisms.METHODS:Forty C57BL mice were randomly divided into control group,MAFLD model group,medium-dose BIH group,and high-dose BIH group.The mice in control group received a regular diet,while those in other groups were fed with high-fat diet for 12 weeks to induce MAFLD.The mice in medium-and high-dose BIH groups received 0.598 and 0.299 g/kg BIH solution,respectively,while those in control and MAFLD groups received an equivalent volume of normal saline.Serum levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were measured using an automatic biochemical analyzer.Liver morphology,steatosis and fibrosis were assessed by HE,oil red O and Masson staining.Levels of TC,tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in liver tissues,and bile acids in serum and ileum tissues were measured by ELISA.Protein expression of FXR and fibroblast growth factor 15(FGF15)in ileum tissues,and FXR,small heterodimer partner(SHP)and cholesterol 7α-hydroxylase(CYP7A1)in liver tissues were analyzed by Western blot.Intestinal microbiota changes were assessed by 16S rRNA gene sequencing.RESULTS:(1)The MAFLD mice exhibited increased serum TC,TG,LDL-C and bile acid levels,liver TC,TNF-α and IL-6 levels,and lipid deposition.However,BIH intervention improved these factors and increased FXR and SHP pro-teins,but decreased CYP7A1 expression in the liver.The protein levels FXR and FGF15 in the ileum were also elevated.(2)Intestinal flora analysis demonstrated that BIH intervention improved the abundance and diversity of intestinal flora in MAFLD mice.Specifically,there was an increase in Bacteroidetes/Firmicutes ratio and a decrease in Proteobacteria and Verrucomicrobia.At the genus level,abundance of Duncaniella,Muribaculum and Paramuribaculum increased,while He-licobacter decreased.CONCLUSION:Treatment with BIH regulates intestinal flora,decreases FXR levels,enhances CYP7A1 expression,promotes bile acid synthesis,reduces hepatic cholesterol accumulation,and attenuates liver steato-sis and inflammation in MAFLD mice,indicating potential therapeutic effects.

Objective:To discuss the effect of the small ubiquitin-like modifier-specific protease 1(SENP-1)/hypoxia-inducible factor 1α(HIF-1α)pathway on chronic intermittent hypoxia(CIH)-induced vascular endothelial injury in the rats,and to clarify the related mechanism.Methods:The SD rats were randomly divided into control group and CIH group,and then the rats in each group were further divided into 2,4,and 6-week subgroups,and there were 8 rats in each subgroup.The rats in CIH group were exposed to CIH in a CIH chamber to induce CIH and create the obstructive sleep apnea hypopnea syndrome(OSAHS)models,while the rats in control group were exposed to normoxic conditions.The serum and thoracic aorta tissue of the rats in various groups were collected at each time point.HE staining was used to observe the thoracic aorta vascular injury of the rats in various groups;ELISA method was used to detect the levels of nitric oxide(NO),endothelin-1(ET-1),von Willebrand factor(vWF),and thrombomodulin(TM)in serum of the rats in various groups;Western blotting method was used to detect the expression levels of SENP-1,HIF-1α,and vascular endothelial growth factor A(VEGFA)proteins in thoracic aorta tissue of the rats in various groups.In vitro,the aortic endothelial cells(rAECs)of the rats were cultured and infected with SENP-1 shRNA adenovirus(sh-SENP-1)to construct the cell line with low expression of SENP-1.The CIH was used to induce the vascular endothelial cell injury,and the cells were divided into CIH group,CIH+sh-NC group,and CIH+sh-SENP-1 group;control group was set up separately.CCK-8 method was used to detect the proliferation activities of the cells in various groups;ELISA method was used to detect the activities of lactate dehydrogenase(LDH)in the supernatant and the levels of NO,ET-1,malondialdehyde(MDA),and activities of superoxide dismutase(SOD)in the cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells in various groups;Western blotting method was used to detect the expression levels of SENP-1,HIF-1α,and VEGFA proteins in the cells in various groups.Results:With the extension of CIH induction time,compared with control group,the thoracic aorta endothelium in CIH group gradually became rough and significantly thickened,the level of serum NO of the rats in CIH group was decreased(P＜0.05),and the levels of serum ET-1,vWF,and TM,and the expression levels of SENP-1,HIF-1α,and VEGFA proteins in thoracic aorta tissue were increased(P＜0.05).Compared with control group,the proliferation activity of the cells in CIH group was decreased(P＜0.05),the LDH activity in the supernatant,the levels of ET-1,MDA,and the apoptotic rate in the cells were increased(P＜0.05),while the levels of NO and activity of SOD in the cells were decreased(P＜0.05),and the expression levels of SENP-1,HIF-1α,and VEGFA proteins in the cells were increased(P＜0.05).Compared with CIH group,the proliferation activity of cells in CIH+sh-SENP-1 group was increased(P＜0.05),the activity of LDH in the supernatant,the levels of ET-1,MDA,and the apoptotic rate of the cells were decreased(P＜0.05),while the level of NO and activity of SOD in the cells were increased(P＜0.05),and the expression levels of SENP-1,HIF-1α,and VEGFA proteins were decreased(P＜0.05).Conclusion:The SENP-1/HIF-1α pathway is highly activated in the thoracic aorta injury tissue of the rats induced by CIH.Silencing SENP-1 expression can reduce CIH-induced vascular endothelial cell injury,and its mechanism may be related to downregulating the activation level of SENP-1/HIF-1α pathway.

This paper introduces Professor Huang Huang's clinical experience in treating depression with modified classical pre-scriptions of Zhongjing.Professor Huang Huang inherited and summarized Zhongjing's academic thoughts on treating emotional disea-ses,and formulated Jieyu Chufan Recipe by combining Banxia Houpu Decoction with Zhizi Houpu Decoction.He believes that depres-sion is characterized by qi stagnation and phlegm obstruction,as well as internal disturbance of stagnation fire,and formulated a treat-ment method of clearing heat and eliminating restlessness,relieving depression and resolving phlegm.Professor Huang Huang uses Jieyu Chufan Recipe as the basic prescription,and combines relevant classical prescriptions with modifications to treat depression ac-cording to different constitutions and syndromes of patients,achieving satisfactory clinical results.

D-mannose has many functional activities and is widely used in food, medicine, agriculture and other industries. D-mannitol oxidase that can efficiently convert D-mannitol into D-mannose has potential application in the enzymatic preparation of D-mannose. A D-mannitol oxidase (PsOX) was found from Paenibacillus sp. HGF5. The similarity between PsOX and the D-mannitol oxidase (AldO) from Streptomyces coelicolor was 50.94%. The molecular weight of PsOX was about 47.4 kDa. A recombinant expression plasmid pET-28a-PsOX was constructed and expressed in Escherichia coli BL21(DE3). The K_m and k_cat/K_m values of PsOX for D-mannitol were 5.6 mmol/L and 0.68 L/(s·mmol). Further characterization of PsOX showed its optimal pH and temperature were 7.0 and 35 ℃, respectively, while its enzyme activity could be stably remained below 60 ℃. The molar conversion rate of 400 mmol/L D-mannitol by PsOX was 95.2%. The whole cells of PsOX and AldO were used to catalyze 73 g/L D-mannitol respectively. The reaction catalyzed by PsOX completed in 9 h and 70 g/L D-mannose was produced. PsOX showed a higher catalytic efficiency compared to that of AldO. PsOX may facilitate the enzymatic preparation of D-mannose as a novel D-mannose oxidase.
Recombinant Proteins/metabolism* ; Paenibacillus/metabolism* ; Mannose/metabolism* ; Escherichia coli/metabolism* ; Mannitol/metabolism*