Stroke is an important disease that leads to disability and death in humans and has relatively high disabil-ity and mortality rates.Microglial cells,as an important component of the central nervous system,can be induced into dif-ferent phenotypes of M1 and M2 by pathological stimulation and play a dual role of neurotoxicity and neuroprotection.This article reviews the role of microglial cells in the progression of ischemic stroke and explores the new targets for the treat-ment of ischemic stroke through intervention of microglial cells.

Objective:To explore the efficacy and safety of Ganhai Weikang capsule (GWC) in the treatment of functional dyspepsia (FD).Methods:A randomized, double-blind, placebo-controlled parallel, multi-center, superiority clinical trial was conducted. From March 2018 to April 2020, totally 324 patients with dyspepsia symptoms, who were diagnosed as chronic non-atrophic gastritis by endoscopy and pathology and met the Rome Ⅳ diagnostic criteria for FD from 7 top hospitals were enrolled, including the First Affiliated Hospital of Naval Medical University (Shanghai Changhai Hospital), Heilongjiang Hospital of Traditional Chinese Medicine, Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital, Qilu Hospital of Shandong University, the First Affiliated Hospital of Zhejiang University, Beijing Hospital of Traditional Chinese Medicine of Capital Medical University and the Third Xiangya Hospital of Central South University. The patients were randomly divided into the GWC group and the placebo group according to the ratio of 1∶1. The patients of GWC group were given GWC and the patients of placebo group were given GWC capsule simulant. The patients of both groups orally took capsules before meals, 2.4 g each time and 3 times per day, and the course of treatment was 4 weeks. The main efficacy index was the total clinical effective rate after 4 weeks, and the secondary efficacy index was the changes of clinical symptom scores of upper abdominal pain, upper abdominal burning, postprandial fullness and early satiety. The safety index included laboratory tests and adverse events. Chi-square test and Wilcoxon rank sum test were used for statistical analysis.Results:A total of 320 FD patients were enrolled in the full analysis set (FAS), which included 161 cases in GWC group and 159 cases in placebo group. A total of 298 cases were in the per-protocol set (PPS), 149 cases each in GWC group and placebo group. The results of FAS and PPS both showed that the total clinical effective rates of the GWC group were higher than those of the placebo group (84.5%, 136/161 vs. 44.0%, 70/159 and 83.9%, 125/149 vs. 46.3%, 69/149), and the differences were statistically significant ( χ2=57.07 and 46.32, both P<0.001). In addition, the differences of the total score of main symptoms and each symptom (upper abdominal pain, upper abdominal burning, postprandial fullness and early satiety) before and after treatment of GWC group were all higher than those of the placebo group (FAS: 10 (7, 14) vs. 5 (3, 11); 3 (2, 4) vs. 2 (0, 3); 2 (0, 4) vs. 1 (0, 3); 3 (1, 4) vs. 2 (1, 3); 2 (0, 4) vs. 1 (0, 3). PPS: 10 (7, 13) vs. 5 (3, 11); 3 (2, 4) vs. 2 (0, 3); 2 (0, 4) vs. 1 (0, 2); 3 (1, 4) vs. 2 (1, 3); 2 (0, 4) vs.1 (0, 3)), and the differences were statistically significant (FAS: Z=5.80, 5.91, 3.19, 3.72 and 3.30; PPS: Z=5.14, 5.11, 2.86, 3.21 and 2.84; all P<0.01). The results of FAS and PPS indicated that the improvement rates of main symptoms and each symptom (upper abdominal pain, upper abdominal burning, postprandial fullness and early satiety) of GWC group were all higher than those of the placebo group (FAS: 77.8% (54.6%, 91.3%) vs. 42.9% (28.6%, 61.5%); 100.0% (60.0%, 100.0%) vs. 50.0% (25.0%, 60.0%); 100.0% (50.0%, 100.0%) vs. 50.0% (25.0%, 100.0%); 71.4% (33.3%, 100.0%) vs. 41.4% (25.0%, 66.7%); 100.0% (50.0%, 100.0%) vs. 50.0% (20.0%, 100.0%). PPS: 77.8% (54.2%, 89.5%) vs. 44.0% (28.6%, 65.0%); 100.0% (60.0%, 100.0%) vs. 50.0% (25.0%, 100.0%); 100.0% (50.0%, 100.0%) vs. 50.0% (25.0%, 100.0%); 71.4% (33.3%, 100.0%) vs. 46.4% (25.0%, 66.7%); 100.0% (50.0%, 100.0%) vs. 50.0% (20.0%, 100.0%)), and the differences were statistically significant (FAS: Z=8.60, 7.72, 4.98, 4.24 and 5.61; PPS: Z=7.90, 7.03, 4.49, 3.88 and 4.83; all P<0.001). After 2 weeks of treatment, the differences of the total score of main symptoms and score of each symptom (upper abdominal pain, upper abdominal burning and early satiety) before and after treatment of GWC group were all higher than those of the placebo group (5.0 (3.0, 8.0) vs. 4.0 (2.0, 6.0); 2.0 (1.0, 2.0) vs. 2.0 (0.0, 2.0); 1.5 (0.0, 2.0) vs. 1.0 (0.0, 2.0); 1.5 (0.0, 2.0) vs. 1.0 (0.0, 2.0)), and the differences were statistically significant ( Z=2.95, 3.44, 2.43 and 2.79, all P<0.05). There was no significant difference in the incidence of adverse events between the GWC group and the placebo group (0.6%, 1/163 vs. 0, 0/159). Conclusion:The clinical total effective rate of GWC in the treatment of FD is superior to that of placebo and it has good safety.

Objective By observing the changes of angiogenesis related factors and cerebral perfusion before and after treatment in patients with large area cerebral infarction，the formation of collateral circulation of Salvia miltiorrhiza polyphenolic acid injection and its effect on the improvement of nerve function were discussed.Methods Forty-four patients with large area cerebral infarction were selected，and all subjects were randomly divided into the salvia miltiorrhiza polyphenolic acid treatment group and the conventional treatment group （control group） according to the ratio of 1：1，with sample content of 22 patients in each group. Both groups were treated with cerebral infarction dehydration，cranial pressure reduction and brain protection，and the salvia miltiorrhiza polyphenolic acid treatment group was treated with Salvia miltiorrhiza polyphenolic acid injection on the basis of the above medication. The course of treatment in both groups was 14 days. The changes of VEGF，bFGF，Ang and quasi-continuous arterial spin labeling were observed in the two groups before and 14 days after treatment. Changes in the ratio of cerebral blood flow （CBF） to CBF in the region of interest （ROI） and contralateral image region （CBF lesions/CBFnormal），as well as changes in NIHSS score and mRS score.Results Serum levels of VEGF，bFGF and Ang in the two groups increased after treatment compared with those before treatment，and the difference was statistically significant （P<0.05）.Fourteen days after treatment，serum levels of VEGF，bFGF and Ang in the salvia miltiorrhiza polyphenolic acid treatment group were significantly higher than those in the control group，with statistically significant differences（P<0.05）.After treatment，CBF lesions/CBFnormal in the two groups were increased compared with those before treatment，and the difference was statistically significant（P<0.05）.Fourteen days after treatment，CBF lesions/CBFnormal in the salvia miltiorrhiza polyphenolic acid treatment group were significantly higher than those in the control group，with statistically significant differences（P<0.05）.The NIHSS and mRS scores of the two groups decreased after treatment compared with before treatment，with statistically significant differences（P<0.05），and the salvianolic acid treatment group decreased significantly compared with the control group （P<0.05）.Conclusion Salvia miltiorrhiza polyphenolic acid for injection can promote angiogenesis in patients with large area cerebral infarction，and can significantly improve cerebral blood flow in diseased brain tissue，improve cerebral perfusion，and then effectively improve the symptoms of neurological impairment

White matter hyperintensities (WMHs) are one of the MRI markers of cerebral small vessel disease, which are more common in the elderly, and are closely associated with the clinical manifestations such as cognitive impairment and gait disorder, however, the pathophysiological mechanism is currently unclear. This article expounds the possible pathophysiological mechanisms of WMHs from many aspects, such as hypoperfusion, blood-brain barrier destruction, oxidative stress, inflammation, and genetics.

Speaking lesson is an effective way and teaching skill to prepare lessons for the teaching of anatomy. Taking the speaking lesson design of lecture teaching of the neck as an example, this article shows the teaching of topographic anatomy for "5+3" integrated clinical medical students. Adhering to the teaching concept of "student-centered", we have launched the theoretical teaching of "problem-centered discussion" and the experimental teaching of "operator responsibility system" in groups, with the "thanksgiving" humanistic education running through the whole process of theoretical and experimental courses. The teaching is a good way to cultivate the students' gratitude consciousness and humanistic quality, self-learning ability, critical thinking ability, communication and expression ability, hands-on operation ability and teamwork spirit, which has laid a solid foundation for training medical professionals in the new era.

White matter hyperintensities (WMHs) is a magnetic resonance imaging phenotype of cerebral small vessel disease,which is manifested as diffuse or confluent subcortical white matter.There is usually no clinical manifestation in the early stage of WMHs,and its pathophysiological mechanism has not been fully elucidated.Studies have shown that a variety of factors are associated with WMHs.The article reviews the risk factors for WMHs.

Objective To evaluate the treatment of posterior malleolar two-part fractures complicated with medial and lateral malleolar fractures via a combination of posterolateral and posteromedial approaches.Methods From January 2014 to January 2017,26 patients were operatively treated at Department of Orthopaedics,Tongji Hospital for posterior malleolar two-part fractures complicated with medial and lateral malleolar fractures via a combination of posterolateral and posteromedial approaches.They were 10 men and 16 women,aged from 53 to 67 years(average,61.5 years).The surgery was conducted in prone position via the posterolateral and posteromedial approaches to expose simultaneously the fractures ends at medial,lateral and posterior malleoli for open reduction.The lateral malleolar fractures were fixated with plate,the medial malleolar fractures with screws and posterior malleolar fractures with plate or cannulated screws depending on the size of the fracture blocks.The outcomes were assessed using the ankle-hindfoot scores of American Orthopaedic Foot and Ankle Society(AOFAS) and the visual analogue scale(VAS).Results Of this cohort,22 were followed up for 30 months on average(range,from 18 to 48 months).All the cases healed by the first intension without any infection.Their postoperative X-ray showed bone union after an average of 12.5 weeks(range,from 10 to 15 weeks).No nonunion,loosening or breakage of implants was found.The mean time for walking with full weight-bearing was 13 weeks(range,from 11 to 16 weeks).Their AOFAS ankle-hindfoot scores at the final follow-ups were 85.4(range,from 80 to 92),yielding 13 excellent and 9 good cases with a good to excellent rate of 100%.Their mean VAS scores were decreased significantly from preoperative 8.6±0.6 to postoperative 1.7±0.3(f=153.000,P=0.000).Conclusion In treatment of posterior malleolar two-part fractures complicated with medial and lateral malleolar fractures,a combination of posterolateral and posteromedial approaches in prone position can expose and reduce simultaneously the fractures ends at medial,lateral and posterior malleoli,leading to satisfactory clinical outcomes.

Objective To investigate the effect of acidic tumor microenvironment on invasion and migration and its mechanism in glioma cells. Methods (1) The pH value of the medium was adjusted by acid-base titration. Human glioma cells U87 and U251 were cultured in the acid group and the normal group with pH values of 6.4 and 7.4, respectively; and 3 d after cultivation, the expressions of hypoxia-inducible factor-2α (HIF-2α) and CD44 were detected by Western blotting; Transwell assay was used to examine the invasion and migration of U87 and U251 cells; immunofluorescence was employed to examine the CD44 expression. (2) The U87 and U251 cells were divided into small interfering RNA (siRNA) -nonsense sequence group and siRNA-CD44-1 group, and the siRNA nonsense sequences and siRNA-CD44-1 interfering fragments were transfected by lipofectin-3000, respectively; three d after transfection, the migration and invasion abilities of cells from the two groups were detected by Transwell assay. (3) U87 and U251 cells were divided into acid group (cultured with a pH value of 6.4), blank control group, siRNA nonsense sequence group, siRNA-CD44-1 group, and siRNA-CD44-2 group; and cells from the later four groups were cultured with a pH value of 7.4; after culture for 4 d, the siRNA-nonsense sequence group, siRNA-CD44-1 group and siRNA-CD44-2 group were transfected with siRNA-nonsense sequences, siRNA-cd44-1 interfering fragments and siRNA-CD44-2 interfering fragments, respectively; three d after transfection, the expressions of CD44, N-Ca, Vimentin, and matrix metalloproteinase (MMP)-2 proteins in these 5 groups were detected by Western blotting. Results (1) As compared with the normal group, the expression levels of HIF-2α and CD44 in U87 and U251 cells of the acid group were significantly increased; both Transwell and invasion experiments showed that the number of transmembrane cells in the acid group was significantly larger than that in the normal group (P<0.05); immunofluorescence staining showed that the CD44 expression in acid group was significantly higher than that in normal group (P<0.05). (2) Both Transwell and invasion experiments showed that the number of transmembrane cells in the siRNA-CD44-1 group was significantly smaller than that in the siRNA nonsense sequence group (P<0.05). (3) Western blotting showed that the expression levels of CD44, N-Ca, Vimentin and MMP-2 in U87 and U251 cells of the blank control group, siRNA nonsense sequence group, siRNA-CD44-1 group, and siRNA-CD44-2 group were obviously decreased as compared with those in the acid group; the expression levels of CD44, N-Ca, Vimentin and MMP-2 in U87 and U251 cells of the siRNA-CD44-1 group and siRNA-CD44-2 group were obviously lower than those in the siRNA nonsense sequence group. Conclusion Acidic tumor microenvironment enhances the capabilities of invasion and migration of glioma cells through increasing CD44 expression.

Objective To study the influence of micro (miR)-325 in progression of glioma and its molecular mechanism by regulating transferrin receptor (TFRC) gene expression in glioma cells. Methods (1) Thirty-five glioma tissues and paired adjacent normal tissues were collected during surgical excision performed in our hospital from January 2015 to January 2018. The miR-325 and TFRC mRNA expression levels in the glioma tissues and paired adjacent normal tissues were detected by inverse transcription-quantitative PCR (RT-qPCR); the expression of miR-325 in glioma tissues of patients with different clinical characteristics and the survival curves of patients with low or high miR-325 expressions were compared. (2) RT-qPCR was used to examine the miR-325 expression in HA, U251, and U87 cell lines in vitro; the regulatory relations between miR-325 and its potential target gene TFRC in U251, and U87 cell lines were measured by luciferase report assay; miR-325 mimic and its negative control were transfected into U251 and U87 cell lines for 48 h, and then, the mRNA and protein expressions of TFRC were detected by RT-qPCR and Western blotting, respectively; control small interfering RNA (siRNA)+nonsense inhibitor, TFRC siRNA+nonsense inhibitor, and siTFRC+miR-325 inhibitor were transfected into U251 and U87 cell lines for 48 h, respectively, Western blotting was employed to detect the TFRC protein expression, cell proliferation was detected by CCK-8 assay, and cell invasion was detected by Transwell assay; pcDNA3.1 empty vector+nonsense sequence, TFRC pcDNA3. 1+nonsense sequence, TFRC pcDNA3.1+miR-325 mimic were transfected into U251 and U87 cell lines for 48 h, respectively, TFRC protein expression was detected by Western blotting, cell proliferation was detected by CCK-8 assay, and cell invasion was detected by Transwell assay. Results (1) As compared with those in the adjacent tissues, the miR-325 expression was significantly decreased and the TFRC mRNA expression was statistically increased in glioma tissues (P<0.05); the TFRC mRNA expression and miR-325 expression were negatively correlated in glioma tissues (P<0.05); as compared with patients with Karnofsky functional status scores≥80, patients with scores<80 had significantly decreased miR-325 expression; as compared with glioma tissues of WHO grading I-II, glioma tissues of grading III-IV had significantly decreased miR-325 expression (P<0.05); the survival rate of patients with low miR-325 expression was statistically lower than that of patients with high miR-325 expression (P< 0.05). (2) As compared with that in HA cells, the miR-325 expression was statistically down-regulated in U87 and U251 cells (P<0.05); in TFRC wild-type (TFRC WT) transfected cells, the miR-325 mimic group had significantly lower luciferase activity than the nonsense sequence group, while the miR-325 inhibitor group had significantly higher luciferase activity than the nonsense inhibitor group (P<0.05); as compared with those in the nonsense sequence group, the TFRC mRNA and protein expressions were statistically decreased in U87 and U251 cells of miR-325 mimic group; as compared with those in the control siRNA+nonsense inhibitor group, the TFRC protein expression and absorbance value were significantly decreased, and number of invasive cells was significantly smaller in the siTFRC+nonsense inhibitor group; and as compared with those in the siTFRC+nonsense inhibitor group, the TFRC protein expression and absorbance value were significantly increased, and number of invasive cells was significantly larger in the siTFRC+miR-325 inhibitor group (P<0.05); as compared with the pcDNA3.1 empty vector+nonsense sequence group, the TFRC protein expression and absorbance value were significantly increased, and number of invasive cells was significantly larger in the TFRC pcDNA3.1 +nonsense sequence group, and as compared with the TFRC pcDNA3.1+nonsense sequence group, the TFRC protein expression and absorbance value were significantly decreased, and number of invasive cells was significantly smaller in the TFRC pcDNA3.1+miR-325 mimic group (P<0.05). Conclusion The miR-325 expression is decreased in glioma cells and has a tumor suppressor effect; patients with low miR-325 expression have poor prognosis; miR-325 inhibits cancer cell progression by inhibiting the expression of the target gene TFRC.

BACKGROUND/AIMS: This study was designed to investigate the effect of Fengliao-Changweikang (FLCWK) in diarrhea-predominant irritable bowel syndrome (IBS-D) rats and explore its underlying mechanisms. METHODS: IBS-D model rats were induced by neonatal maternal separation (NMS) combined with restraint stress (RS). In in vivo experiments, the model rats were randomly divided into 5 groups: NMS + RS, FLCWK (low dose, middle dose, and high dose), and pinaverium bromide. The normal control (no handling) rats were classified as the NH group. The therapeutic effect of FLCWK was evaluated by fecal characteristics, electromyographic response and abdominal withdrawal reflex scores. In in vitro experiments, the model rats were randomly divided into 2 groups: NMS + RS, FLCWK (middle dose), and no handling rats were used as the NH group. The differences in basic tension and ACh-induced tension of isolated colonic longitudinal smooth muscle strips (CLSMs) among the 3 groups were observed. In addition, different inhibitors (nifedipine, TMB-8, L-NAME, methylene blue, and 4-AP) were pretreated to explore the underlying mechanisms. RESULTS: In in vivo experiments, fecal characteristics, electromyographic response, and abdominal withdrawal reflex scores significantly improved in the FLCWK group, compared with the NMS + RS group. In in vitro experiments, the basic tension and ACh-induced tension of CLSMs in IBS-D rats were significantly inhibited by FLCWK. After pre-treatment with different inhibitors, the ACh-induced tension of CLSMs in each group showed no significant difference. CONCLUSIONS: FLCWK manifested curative effect in IBS-D rats by inhibiting colonic contraction. The underlying mechanisms may be related to regulatory pathway of nitric oxide/cGMP/Ca2+ and specific potassium channels.
Animals ; Colon ; Gastrointestinal Motility ; Herbal Medicine ; In Vitro Techniques ; Irritable Bowel Syndrome ; Methylene Blue ; Muscle, Smooth ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Potassium Channels ; Rats ; Reflex