Objective: To investigate the potential mechanism of Maxing Shigan Decoction (MSD) in the prevention and treatment of influenza virus infection by influencing pulmonary flora and expression of chemokines CCL5 and CXCL10 of mice. Method The infected mice model of influenza A virus was tested by intranasal inoculation. After 3 and 7 d of gavage or saline, the lung index and lung index inhibition rate were calculated. Pathological changes of lung tissue were detected by HE staining. The expression of CCL5 and CXCL10 in the lung tissue of mice was detected by immunohistochemistry and ELISA. The expression of CCL5 mRNA and CXCL10 mRNA in lung tissue of mice was detected by real-time fluorescence quantitative PCR (RT-PCR). The bacteria in lung tissue was sequenced by using the V3-V4 variable region of 16S rRNA, annotated and clustered. The alpha diversity, beta diversity and the species difference among groups were analyzed. The correlation of the expression of CCL5 and CXCL10 with the change of intestinal flora was also analyzed. Results: After 3 d of administration, the lung index of model group was significantly higher than normal group (P < 0.01) and drug group (P < 0.05, 0.01). Pulmonary inflammatory cell infiltration was obvious. The infiltration of pulmonary inflammatory cells in MSD group was significantly reduced, and the inhibition rate of lung index was similar to that in oseltamivir group. The value of IQA in lung injury was decreased significantly (P < 0.01). The expressions of CCL5 and CXCL10 in the lung tissue of the model control group were significantly higher than those of the normal control group (P < 0.01), and the expressions of CCL5 and CXCL10 in the oseltamivir group and the MSD were significantly lower than those in the model control group (P < 0.05, 0.01). The results of 16S rRNA gene sequencing showed the relative abundances of Bacteroides, Escherichia, and Proteus were increased, while that of Coprococcus was decreased in the model control group. In oseltamivir group and MSD group, the relative abundances of Bacteroides, Escherichia, and Proteus were significantly decreased, while the relative abundance of Coprococcus was increased. The results of alpha diversity showed that the ace index, Chao1 index, and Shannon index of each group were all higher than 0.05, and there was no difference in richness and diversity among groups. The results of beta diversity showed that there was no intersection of sample points among groups and difference in the composition of pulmonary flora among groups. Species among groups were significant differences. Spearman correlation analysis showed that the expression of CCL5 and CXCL10 was positively correlated with the abundance of Escherichia, Proteus, and Bacteroides, and negatively correlated with the abundance of Coprococcus. After 7 d of administration, there was no significant difference in the composition of pulmonary flora and the expression of CCL5 and CXCL10. Conclusion: MSD may improve the micro-ecological environment and immune microenvironment of the lung by promoting the growth of beneficial bacteria, and has a certain protective effect on the lung injury caused by influenza virus.

Classical prescription is long-lasting which is characterized by simple composition and obvious curative effect. It is popular over China and welcomed by many people. Japanese Kampo medicine and traditional Chinese medicine originated from the same resource but flowed into two branches. Some classical prescription experts are mostly influenced by Japanese Kampo medicine, emphasizing the correspondence between prescriptions and syndromes, physical identification and abdominal diagnosis, which have become the research direction of most classical prescription scholars. Traditional Chinese medicine and Japanese Kampo medicine differ in the dialectical system, application form, dose and dosage form of classical prescription. It is important to have a clear insight of the development status of classical prescription in Chinese medicine and that of Japanese Kampo medicine, understand the differences between them and make full use of advantages and avoid disadvantages. It can provide ideas for the correct development path of classical prescription and reference for the research of preparation of classical prescription in China.

OBJECTIVETo reveal the effects and related mechanisms of chlorogenic acid (CGA) on intestinal glucose homeostasis.

METHODSForty male Sprague-Dawley rats were randomly and equally divided into four groups: normal chow (NC), high-fat diet (HFD), HFD with low-dose CGA (20 mg/kg, HFD-LC), and HFD with high-dose CGA (90 mg/kg, HFD-HC). The oral glucose tolerance test was performed, and fast serum insulin (FSI) was detected using an enzyme-linked immunosorbent assay. The mRNA expression levels of glucose transporters (Sglt-1 and Glut-2) and proglucagon (Plg) in different intestinal segments (the duodenum, jejunum, ileum, and colon) were analyzed using quantitative real-time polymerase chain reaction. SGLT-1 protein and the morphology of epithelial cells in the duodenum and jejunum was localized by using immunofluorescence.

RESULTSAt both doses, CGA ameliorated the HFD-induced body weight gain, maintained FSI, and increased postprandial 30-min glucagon-like peptide 1 secretion. High-dose CGA inhibited the HFD-induced elevation in Sglt-1 expression. Both CGA doses normalized the HFD-induced downregulation of Glut-2 and elevated the expression of Plg in all four intestinal segments.

CONCLUSIONAn HFD can cause a glucose metabolism disorder in the rat intestine and affect body glucose homeostasis. CGA can modify intestinal glucose metabolism by regulating the expression of intestinal glucose transporters and Plg, thereby controlling the levels of blood glucose and insulin to maintain glucose homeostasis.

Animals ; Chlorogenic Acid ; pharmacology ; Diet, High-Fat ; adverse effects ; Glucagon-Like Peptide 1 ; metabolism ; Glucose ; metabolism ; Glucose Tolerance Test ; Glucose Transporter Type 2 ; metabolism ; Homeostasis ; Insulin ; blood ; Intestines ; drug effects ; metabolism ; Male ; Proglucagon ; metabolism ; Random Allocation ; Rats, Sprague-Dawley ; Sodium-Glucose Transporter 1 ; metabolism ; Weight Gain ; drug effects

Background Silicosis is a diffuse fibrosis of the lungs caused by long-term inhalation of free silicon dioxide (SiO₂). It has a complex pathogenesis and lacks effective treatment. Brusatol (Bru) has a variety of biological activities, and its role in silicosis fibrosis is unclear yet. Objective To investigate the effects of different concentrations of Bru on SiO₂-induced silicosis fibrosis in mice. Methods Thirty male C57BL/6J mice were randomly divided into five groups: a control group, a silica group, and three Bru intervention groups with low, medium, and high doses (1, 2, and 4 mg·kg⁻¹), with 6 mice in each group. Except the control group, the remaining groups were established as SiO₂-induced silicosis mouse models by using a single tracheal infusion of 50 μL 60 mg·mL⁻¹ SiO₂ suspension. The control group was dosed with equal amount of saline. The Bru intervention groups were injected intraperitoneally with Bru for 5 consecutive days and then injected every other day. After 28 d of exposure, the mice were executed and lung tissues were collected. The lung coefficient of the mice was measured, and the pathological changes of the lung tissues were observed after hematoxylin-eosin (HE) and Masson staining. The levels of apoptotic protein Cleaved-caspase 3, fibrosis-related protein α-smooth muscle actin (α-SMA), type I collagen (Col-I), autophagy-associated protein Beclin1, microtubule-associated protein 1 light chain 3 (LC3), Sequestosome 1 (p62/SQSTM1), Kelch like ECH-associated protein-1 (Keap1), and nuclear factor erythroid 2 related factor 2 (Nrf2) were detected by Western blot. The mRNA levels of Caspase 3, α-SMA, and Col-I were measured by realtime fluorescence-based quantitative PCR. Results Compared with the control group, the lung coefficient of mice in the silica group was significantly increased (P < 0.01); the lung tissues of the silicosis mice showed damaged alveolar walls, along with infiltration of inflammatory cells, fibrous nodules, and collagen deposition; furthermore, the protein and mRNA levels of Cleaved-caspase 3, α-SMA, and Col-I were significantly increased (P < 0.01); the expression levels of Beclin1, LC3-II/I, p62, and Nrf2 were increased, while that of Keap1 was decreased (P < 0.05). The interventions with low and medium doses of Bru reduced lung coefficient (P < 0.05) and protected against pathological damage and collagen deposition in the lung tissues of the silicosis mice; the protein and mRNA expression levels of Cleaved-caspase 3, α-SMA, and Col-I were significantly decreased in the low and medium dose groups (P < 0.05, P < 0.01), the expression levels of Beclin1, LC3-II/I, p62, and Nrf2 were also decreased (P < 0.05, P < 0.01), and the expression level of Keap1 was increased in the medium dose group (P < 0.05). However, compared with the silica group, the differences in lung coefficient, pathological damage, and protein and mRNA expression levels of Cleaved-caspase 3, α-SMA, and Col-I in the Bru high dose group were not statistically significant (P > 0.05). In addition, the high dose of Bru decreased Beclin1, LC3-II/I, and Nrf2 expression levels (P < 0.01), did not change p62 protein expression level (P > 0.05), while increased Keap1 protein level (P < 0.01). Conclusion Low and medium doses of Bru might regulate autophagy through the Keap1-Nrf2 pathway, ameliorate autophagic degradation impairment, reduce pulmonary coefficient, attenuate apoptosis, and delay the progression of fibrosis in SiO₂-induced silicosis mice.

Mesenchymal stem cell (MSCs) has recently emerged as an appealing and potential therapeutic strategy to cure a diverse range of diseases in the orthopaedic field. Owing to its capacity of osteogenic differentiation, most of researches just focused on promoting MSC differentiation. With the in-depth study, MSCs homing is also a key issue for bone formation and bone diseases treatment, which have been described that MSCs mobilize from in situ environment (bone marrow) and migrate into injured tissues during the healing process through peripheral circulation. MSC homing is the incipient step of bone formation. MSCs need to firstly migrate to the bone surface and then differentiate into osteogenic cells to enhance bone repair. Promoting MSCs homing have been shown to improve recovery of several orthopedic diseases, such as osteoporosis, fracture, bone defect and wear-particle-related osteolysis. Therefore, further research on MSCs homing may provide a new thinking for treatment of osteoporosis.
Bone Marrow ; Cell Differentiation ; Humans ; Mesenchymal Stem Cells ; Osteogenesis ; Osteoporosis

OBJECTIVE: To observe the effects of Taohong Siwu Decoction(, THSWD) on the mesenchymal stem cells(MSCs) migration, homing number and cytokine expression in callus during the early process of fracture healing, and to explore the mechanism of THSWD on accelerationg fracture healing by regulating the homing of MSCs in rats. METHODS: A rat model of right femoral shaft open fracture was established. Thirty-two 5-week-old male Sprague-Dawley rats, weighting 110 to 130 g, were divided into control group, low-dose group, medium-dose group and high-dose group by using random number table. Distilled water was given to the control group, and the other groups were given Taohong Siwu Decoction. The rats were gavaged twice a day for 5 consecutive days after surgery. Bone volume/tissue volume(BV/TV) and bone mineral density(BMD) were observed using micro-computed tomography (micro-CT) at 21 days after surgery. At 5 days post-fracture, peripheral blood MSCs from THSWD treated and untreated rats were cultured in vitro. Subsequently, the migration ability of MSCs was observed by cell migration assay. The number of MSCs homing to the callus at the early stage of fracture (5 d) was detected by Immunohistochemistry (IHC). Protein chip was used to detect the expression of cytokines in callus. RESULTS: Micro-CT results showed that BV/TV was higher in the high-dose group than in the medium-dose group (P=0.032), and higher in the medium-dose group than in the low-dose group(P=0.041), with no difference between the control and low-dose group (P=0.651). In addition, there was no difference in BMD between low-dose group and the model group (P=0.671), and lower in the low-dose group than in the medium-dose group(P=0.018), and the medium-dose group was lower than the high-dose group(P=0.008). Cell migration assay showed that THSWD promotes enhanced the migration ability of peripheral blood MSCs. IHC assay revealed that CD45^-, CD90⁺, CD29⁺ MSCs significantly increased in bone callus after THSWD intervention compared with the control group. Protein chip showed that THSWD promoted the upregulation of CINC-1(×2.91), CINC-3(×1.59), LIX(×1.5), Thymus Chemokine (×2.55), VEGF (×1.22) and the down-regulation of TIMP-1 (×2.98). CONCLUSION THSWD, a representative formula of "promoting blood circulation and removing blood stasis", can significantly accelerate fracture healing, and its mechanism may be related to enhancing the migration ability of peripheral blood MSCs and up-regulating CINC-1, CINC-3, LIX, Thymus Chemokine, VEGF and down-regulating TIMP-1 in bone callus, which promotes the peripheral blood MSCs homing in the early stage of fracture.
Animals ; Drugs, Chinese Herbal ; Fracture Healing ; Fractures, Bone/drug therapy* ; Humans ; Male ; Mesenchymal Stem Cells ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1/pharmacology* ; Vascular Endothelial Growth Factor A ; X-Ray Microtomography

Objective : To explore the mechanism of Wenyang Shengji Ointment (温阳生肌膏, WYSJO) in the treatment of diabetic wounds from the perspective of network pharmacology, and to verify it by animal experiments. Methods: The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and related literature were used to screen active compounds in WYSJO and their corresponding targets. GeneCards, Online Mendelian Inheritance in Man (OMIM), DrugBank, PharmGkb, and Therapeutic Target Database (TTD) databases were employed to identify the targets associated with diabetic wounds. Cytoscape 3.9.0 was used to map the active ingredients in WYSJO, which was the diabetic wound target network. Search Tool for the Retrieval of Interaction Gene/Proteins (STRING) platform was utilized to construct protein-protein interaction (PPI) network. Kyoto Encyclopedia of Genes and Genomes (KEGG) andGene Ontology (GO) enrichment analyses were performed to identify signaling pathways between WYSJO and diabetic wounds. AutoDock 1.5.6 was used for molecular docking of core components in WYSJO to their targets. Eighteen rats were randomly divided into control, model, and WYSJO groups (n = 6). The model and WYSJO groups were used to prepare the model of refractory wounds in diabetes rats. The wound healing was observed on day 0, 5, 9, and 14 after treatment, and the wound tissue morphology was observed by hematoxylin-eosin(HE) staining. The expression levels of core genes were detected by quantitative real-timepolymerase chain reaction (qPCR). Result: A total of 76 active compounds in WYSJO, 206 WYSJO drug targets, 3 797 diabetic wound targets, and 167 diabetic wound associated WYSJO targets were screened out through network pharmacology. With the use of WYSJO-diabetic wound target network, core targets of seven active compounds encompassing quercetin, daidzein, kaempferol, rhamnetin, rhamnocitrin, strictosamide, and diisobutyl phthalate (DIBP) in WYSJO were found. GO enrichment analysis showed that the treatment of diabetes wounds with WYSJO may involve lipopolysaccharide, bacteria-derived molecules, metal ions, foreign stimuli, chemical stress, nutrient level, hypoxia, and oxidative stress in the biological processes. KEGG enrichment analysis showed that the treatment of diabetes wounds with WYSJO may involve advanced glycation end products (AGE-RAGE), p53, interleukin (IL)-17, tumor necrosis factor (TNF),hypoxia inducible factor-1 (HIF-1), apoptosis, lipid, atherosclerosis, etc. The results of animal experiments showed that WYSJO could significantly accelerate the healing process of diabetic wounds (P < 0.05), alleviate inflammatory response, promote the growth of granulation tissues, and down-regulate the expression levels of eight core genes [histone crotonyltransferase p300 (EP300), protoc gene-oncogene c-Jun (JUN), myelocytomatosis (MYC), hypoxia inducible factor 1A (HIF1A), mitogen-activated protein kinase 14 (MAPK14), specificity protein 1 (SP1), tumor protein p53 (TP53), and estrogen receptor 1 (ESR1)] predicted by the network pharmacology (P < 0.05). Conclusion The mechanism of WYSJO in treating diabetes wounds may be closely related to AGE-RAGE, p53, HIF-1, and other pathways. This study can provide new ideas for the pharmacological research of WYSJO, and provide a basis for its further transformation and application.

OBJECTIVE: To evaluate clinical effects of Double-pulley dual row technique with shoulder arthroscopy in treating scapular glenoid fracture(Ideberg typeⅠ). METHODS: From July 2017 to March 2019, 8 patiens with scapular glenoid fracture (Ideberg typeⅠ) were treated with Double-pulley dual-row technique with shoulder arthroscopy, including 7 males and 1 female;5 cased of injuries in the left shoulder, 3 cased of injuries in the right shoulder;ranging in age from 22 to 56 years old; and the time from injury to operation ranged from 3 to 10 days. X-ray and CT of shoulder joint were taken before and after operation to evaluate the fracture severity and fracture healing. American Shoulder and Elbow Surgeous (ASES) and Constant- Murley scores were used to evaluate shoulder joint function. RESULTS: All patients were followed up, and the duration ranged from 12 to 24 months, and the fracture healing time ranged from 3 to 5 months. No operative site infection was found in all patients. CT scan of shoulder joint showed satisfactory reduction and no displacement. The shoulder joint function recovered well. ASES score at the latest follow up after operation ranged from 85 to 97 points, which were higher than those before operation; Constant-Murley score ranged from 83 to 96 points, which were higher than those before operation. CONCLUSION Double-pulley dual-row technique with shoulder arthroscopy is effective to fix scapular glenoid fracture of Ideberg typeⅠwith minimal tissue trauma and significant improvement of shoulder joint function.
Adult ; Arthroscopy ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Scapula/surgery* ; Shoulder ; Shoulder Joint/surgery* ; Treatment Outcome ; Young Adult

C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Hernia, Inguinal ; immunology ; surgery ; Humans ; Infant ; Interleukin-6 ; blood ; Laparoscopy ; methods ; Tumor Necrosis Factor-alpha ; blood

AIM:To study the ocular surface and corneal lesions under different degrees of retinopathy in patients with type 2 diabetes mellitus(T2DM).

METHODS: A total of 123 patients(246 eyes)with T2DM were enrolled in this study. All of patients were divided into non-diabetic retinopathy group(46 patients 92 eyes), non-proliferative diabetic retinopathy group(50 patients 100 eyes)and proliferative diabetic retinopathy group(27 patients 54 eyes)according to the type of retinopathy. Dry eye questionnaire, ocular surface disease index(OSDI), Schirmer Ⅰ text(SⅠt), tear break-up time(BUT), fluorescein staining of cornea(FL), corneal endoscopy and central corneal thickness measurement were used to analyze the differences of three groups.

RESULTS: There was no statistically significant difference in general data between the three groups. Significant differences existed in eye pain, dry eyes, lacrimation, eye fatigue, burning sensation, vision fluctuations in three groups(P<0.05). Foreign body sensation, itchy eyes, red eyes showed no significant difference in groups(P>0.05). There were significant differences in ODSI value, FL positive rate, BUT, SIt, corneal endothelial cell density and central corneal thickness in three groups(P<0.05). variable coefficient of corneal endothelial cell showed no Significant difference in groups(P>0.05).

CONCLUSION: The findings of this study show that patients with type 2 diabetes mellitus presented obviously discomfort symptoms of ocular surface, decreased tear film stability, increased positive rate of corneal fluorescein staining; decreased density of corneal endothelial cells; increased central corneal thickness in diabetic patients, all of which were associated with degree of retinopathy.