ObjectiveTo explore the mechanism of Huoxue Rongluo prescription （HXRLP） in repairing the blood-brain barrier （BBB） after ischemic stroke （IS）. MethodsMale C57BL/6 mice were randomly divided into sham operation （Sham） group， cerebral infarction model （MCAO） group， environmental circadian disruption with cerebral infarction model （ECD-MCAO） group， low-， medium-， and high-dose HXRLP （HXRLP-L， M， and H） groups （8.5， 17， 34 g·kg^-1·d^-1， respectively）， and positive drug butylphthalide （NBP） group （0.23 mL·d^-1）. In the Sham group， only the exposed blood vessels were isolated without suture insertion. In the other groups， the middle cerebral artery occlusion （MCAO） model of mice was prepared. In the ECD-MCAO group， HXRLP groups， and NBP group， the environmental circadian disruption （ECD） model was prepared. The mice in the Sham group， MCAO group， and ECD-MCAO group were given the same volume of soybean oil by gavage， while those in the other groups were given the corresponding drugs by gavage. Samples were collected after 7 consecutive days of administration. The mNSS score was used to evaluate the repair effect of HXRLP on neurological deficits after IS. Hematoxylin-eosin （HE） staining was used to assess the impact of HXRLP on the pathological damage of brain tissue after IS. 2，3，5-Triphenyltetrazolium chloride （TTC） staining and cerebral blood perfusion status were used to evaluate the repair effect of HXRLP on brain tissue damage after IS. Evans blue staining and transmission electron microscopy were used to evaluate the improvement effect of HXRLP on the permeability injury of BBB after IS. Immunofluorescence （IF） staining was used to observe the expression of von Willebrand Factor （vWF）， brain and muscle Arnt-like 1 （Bmal1）， and Occludin in brain tissue. Western blot was used to detect the protein expression of Bmal1， Occludin， tight junction protein （Claudin-5）， vascular endothelial growth factor （VEGF）， and angiopoietins（Ang）， and related analysis was conducted. ResultsCompared with the Sham group， the MCAO group exhibited significantly aggravated neurological deficits， cerebral infarction volume， brain pathological damage， and BBB leakage （P0.01） and significantly reduced cerebral blood perfusion （P0.01）. The expression of Bmal1， vWF， vascular endothelial growth factor A （VEGFA）， and Ang in brain tissue was significantly enhanced （P0.01）， while the expression of Occludin and Claudin-5 was significantly weakened （P0.01）. Compared with the MCAO group， the ECD-MCAO group showed significantly aggravated neurological deficits， cerebral infarction volume， and BBB leakage （P0.01）， obviously worsened brain pathological damage （P0.05）， significantly reduced cerebral blood perfusion （P0.01）， and significantly decreased expression of Bmal1， vWF， VEGFA， Ang， Occludin， and Claudin-5 in brain tissue （P0.01）. Compared with the ECD-MCAO group， the HXRLP groups of all doses presented significantly improved neurological deficits， cerebral infarction volume， brain pathological damage， and BBB leakage （P0.01）， significantly increased cerebral blood perfusion （P0.01）， and enhanced expression levels of Bmal1， vWF， VEGFA， Ang， Occludin， and Claudin-5 in brain tissue （P0.01）. ConclusionHXRLP can regulate the clock protein Bmal1 and promote the expression of VEGFA， Ang， Occludin， and Claudin-5， thereby improving BBB damage after IS.

Objective To investigate the role and action mechanism of rosemarinic acid(RA)on the malignant behavior of colorectal cancer(CRC)SW480 cells.Methods We collected tumor tissues from patients with primary untreated CRC who underwent surgery in People's Hospital of Rugao from January 2019 to December 2020,as well as colon tissues from normal individuals.The expression of β-catenin was detected by immunohistochemistry and Western blotting to analyze its relationship with the clinical stage,prognosis,and immune infiltration.CCK-8 was used to detect the effects of 10,15 and 20 μmol/L of RA on the proliferation of SW480 cells;flow cytometry detected apoptosis;Transwell and scratch assay detected changes in the invasion and migration ability of the cells,respectively.Western blotting detected the expressions of proteins related to apoptosis,epithelial-mesenchymal transition(EMT),and Wnt/β-catenin pathway.A mouse model of CRC transplant tumor was established,and the effect of RA on the growth of transplant tumor was investigated by gavage administration.The levels of inflammatory factors were detected by ELISA.Results The expression of β-catenin was low in normal colon tissues and significantly higher in CRC tissues.In the CRC patients with high expression of β-catenin,the number of patients with tumors larger than 5 cm was significantly higher than that in the low expression group,whereas the overall survival of the patients was significantly smaller than that in the low expression group(P＜0.05).Cell proliferation was significantly inhibited and decreased in a concentration-dependent manner in the RA group at all concentrations(P＜0.05).In animal experiments,the expression of apoptotic proteins Bax and Bad was significantly increased and the expression of anti-apoptotic protein Bcl-2 was significantly decreased in the RA-treated group compared with the control group(P＜0.05).It was also found that the invasive and migratory ability of SW480 cells was significantly inhibited(P＜0.05),the expression of E-cadherin protein was significantly increased,the expression of Snail,N-cadherin and Vimentin was significantly reduced(P＜0.05),the expression of Axin and GSK-3β was increased,and the expression of β-catenin was reduced(P＜0.05).Conclusion RA may inhibit the biological activity of SW480 by interfering with Wnt/β-catenin pathway-mediated EMT.

【Objective】 To analyze the height growth of children aged 6 - 15 years in Nanning, Guangxi Zhuang Autonomous Region, so as to provide evidence for the assessment of local children′s height development. 【Methods】 A total of 25 225 children aged 6 - 15 years were selected to get their physical examination data from 41 primary schools in Nanning by stratified cluster sampling method in December 2021.Then the height data were compared with the current domestic standards. 【Results】 The average height of boys in Nanning was lower than the national standard before the age of 10 years and 7 months, and the gap with the national standard gradually narrowed after the age of 10 years and 7 months. The average height of boys in Nanning City exceeded the national standard between the age of 11 years and 1 month and 13 years and 6 months, and then lagged behind the national standard again after the age of 13 years and 7 months. The mean height of girls in Nanning City was lower than the national standard height in several age groups, and it was more obvious before the age of 9 years and 7 months. The proportion of height ≤-2s,≤-s,≥ +s and ≥+2s in boys aged 6 to 15 years in Nanning City fluctuated from 2.59% to 6.04%, 12.09% to 23.43%, 7.18% to 18.79% and 0.93% to 3.14%, respectively; the total proportions were 4.56%, 17.46%, 11.35% and 1.74%, respectively; the minimum/maximum proportion values of each height group were at 11 years old /8 years old, 14 years old/8 years old, 8 years old/12 years old, and 6 years old/11 years old, respectively. The proportions of girls aged 6 - 15 years in Nanning City with height ≤-2s, ≤-s, ≥+s, and ≥+2s fluctuated from 2.06% to 5.19%, 9.35% to 25.15%, 8.21% to 15.80% and 1.23% to 3.49%, respectively; the total proportions were 3.38%, 16.91%, 11.97% and 2.29%, respectively; and the minimum/maximum proportion values of each height group were at 13 years old/6 years old, 12 years old/6 years old, 7 years old/12 years old, and 6 years old/11 years old, respectively. 【Conclusions】 The overall height level of children in Nanning is still lower than the national level, with short prepubertal basal heights, an earlier age of onset of accelerated pubertal height, and a shorter duration of accelerated pubertal height in boys. Strengthening pre-pubertal height management and emphasizing the onset and duration of children′s pubertal development, especially the height development of boys during puberty, can help improve the adult lifelong height of children in this region.

[Objective] To investigate the inhibitory effect of quercetin on cisplatin-resistant human colorectal cancer (CRC) cells SW480 and SW620 and its possible mechanism. [Methods] Cisplatin-resistant subtypes of SW480 and SW620 cells were first cultured and the effects of quercetin on cell proliferation and chemosensitivity were determined by MTT assay. Flow cytometry was used to detect apoptosis and protein blotting was used to detect the expressions of relevant proteins. [Results] MTT assay showed that quercetin at the concentrations of 80 μmol/L and 160 μmol/L significantly inhibited the proliferation of SW480 and SW620 cells. Flow cytometry results showed that the apoptosis rate was significantly higher in the cisplatin combined with quercetin group than in the other three groups. Protein blotting showed that cleaved-caspase-3 and caspase-9 expressions were significantly increased in the cisplatin combined with quercetin group. The chemotherapeutic drug sensitivity assay showed that the elevated IC50 of drug-resistant cells was decreased significantly after quercetin administration (P<0.05). [Conclusion] The present study clarifies that quercetin can increase the sensitivity of human CRC cells to cisplatin. This effect may be related to its mechanism of action in affecting cell proliferation, apoptosis and autophagy.

Objective To identify lipid metabolism genes in cerebral infarction;To explore the intervention effect of Huoxue Rongluo Prescription.Methods Multi-chip combined differential analysis(GSE61616,GSE30655)was used to identify lipid metabolism genes in cerebral infarction in combination with Reactome database,and the expression differences of lipid metabolism genes in cerebral infarction were identified and verified in GSE97537 chip;Pearson correlation analysis was used to analyze the correlation of 51 cerebral infarction samples in GSE61616,GSE30655,GSE97537,GSE137595,GSE22255,GSE163614,and GSE78731 datasets;PPI,GO and KEGG analysis of lipid metabolism genes in cerebral infarction were performed through STRING database and R clusterProfiler package.SD rats were made to the model of cerebral infarction,and was administered with Huoxue Rongluo Prescription extract 11.7 g/kg by intragastric administration for 7 days.The symptoms of neurological deficit,the changes of Nissl bodies and the mRNA expressions of PLA2G4A,SPHK1,and PTGES key genes in lipid metabolism in cerebral infarction were observed.Results TSPO,CYP1B1,PLIN2,CH25H,PLA2G4A,ANGPTL4,PTGS1,SPHK1,and PTGES were identified as lipid metabolism genes in cerebral infarction,and were significantly highly expressed and positively correlated in cerebral infarction.Among them,PTGS1,PLA2G4A,and SPHK1 interacted with each other,which were the key genes of lipid metabolism in cerebral infarction;the lipid metabolism gene in cerebral infarction mainly exerted molecular functions such as oxidoreductase activity,iron ion binding,heme binding,etc.,mediating arachidonic acid metabolism,phospholipase D signaling pathway,VEGF signaling pathway,involved in regulation of lipid metabolism process,fatty acid metabolism process,fatty acid derivative metabolism process.The symptoms of neurological deficit in the model rats with cerebral infarction were severe(P<0.001),and Huoxue Rongluo Prescription could effectively improve the neurological deficit of model rats(P<0.001).The Nissl staining indicated that the neuronal structure was abnormal and the number was significantly reduced after cerebral infarction(P<0.001).Huoxue Rongluo Prescription could increase the number of neurons(P<0.001)and repair the neuronal structure.RT-qPCR showed that the key genes of lipid metabolism in cerebral infarction were significantly higher in cerebral infarction(P<0.001),corroborated with the bioinformatics results,and Huoxue Rongluo Prescription could reduce the expression of key lipid metabolism genes of PTGS1,PLA2G4A,and SPHK1(P<0.001,P<0.01,P<0.05).Conclusion Huoxue Rongluo Prescription can down-regulate the expressions of PTGS1,PLA2G4A,SPHK1,exert molecular functions such as oxidoreductase activity,iron ion binding,heme binding,and mediate arachidonic acid metabolism,phospholipase D signaling pathway,and VEGF signaling pathway.It participates in the process of lipid metabolism regulation,fatty acid metabolism,and fatty acid derivative metabolism,increases the number of Nissl bodies,improves the symptoms of neurological deficits,and exerts neuroprotective effects.

Objective:To investigate the application of modified Hanley surgery in abscess of anal tube space (AATS), and its influences on anal function, interleukin (IL)-8 and IL-6.Methods:The clinical data of 96 patients with AATS from February 2020 to February 2022 in Shaoxing Central Hospital were retrospectively analyzed. Among them, 48 patients were treated with incision-thread drawing procedure (ITDP group), and 48 patients were treated with modified Hanley group (modified Hanley group). The curative effect, anal function, inflammatory factor level, neovascularization factor level and wound healing status were compared between the two groups.Results:There was no significant difference in the recovery rate between modified Hanley group and ITDP group: 100.00% (48/48) vs. 95.83% (46/48), χ2 = 0.51, P>0.05. The poor rate of anal function 2 months after surgery in modified Hanley group was significantly lower than that in ITDP group: 0 vs. 12.50% (6/48), and there was statistical difference ( P<0.05). The tumor necrosis factor-α(TNF-α), IL-8 and IL-6 24 h after surgery in modified Hanley group were significantly lower than those in ITDP group: (127.11 ± 13.96) ng/L vs. (160.59 ± 11.57) ng/L, (92.20 ± 11.62) ng/L vs. (124.33 ± 12.05) ng/L and (79.38 ± 12.47) ng/L vs. (100.07 ± 12.50) ng/L, and there were statistical differences ( P<0.01). The monocyte chemotactic protein-1 1 week after surgery in modified Hanley group was significantly lower than that in ITDP group: (92.85 ± 14.63) ng/L vs. (122.90 ± 15.59) ng/L, the vascular endothelial growth factor-A and transforming growth factor-β 1 were significantly higher than those in ITDP group: (188.06 ± 22.53) ng/L vs. (137.80 ± 19.52) ng/L and (1 897.6 ± 97.3) ng/L vs. (1 608.6 ± 98.1) ng/L, and there were statistical differences ( P<0.01). The pain score, edema score and neonatal granulation score 7, 14 and 21 d after surgery in modified Hanley were significantly lower than those in ITDP group, and there were statistical differences ( P<0.01). Conclusions:In the treatment of AATS, the modified Hanley surgery not only contributes to the thorough removal of the lesions, but also plays a positive role in protecting the anal function, reducing the level of inflammation, and promoting the recovery of endothelial function.

Objective:To explore the clinical efficacy of small incision floating line drainage for the treatment of high perianal abscess.Methods:A retrospective analysis was conducted on the clinical data of 95 patients with high perianal abscess treated at the Medical Community General Hospital of Shaoxing Central Hospital from April 2019 to April 2021. Among them, 47 cases were treated with small incision floating line drainage (experimental group), and 48 cases were treated with conventional multi incision drainage (control group). The surgical time, intraoperative bleeding, postoperative pain, urination status, anal function evaluation, wound healing status and the clinical efficacy of the patient after 2 months of treatment were compared between the two groups.Results:The surgical time, intraoperative bleeding volume in the experimental group were lower than those in the control group: (18.70 ± 0.48) min vs. (38.10 ± 2.52) min, (32.35 ± 3.56) ml vs. (51.56 ± 6.24) ml, there were statistical differences ( P<0.05). The postoperative pain, urination status, anal function evaluation in the experimental group were better than those in the control group. In the experimental group, multiple incision drainage had a greater impact on patients and the wound healing cycle was longer. After treatment for 2 months, the total effective rate in the experimental group was better than that in the control group: 100.00%(47/47) vs. 91.67%(44/48), there was statistical difference ( χ2 = 4.09, P<0.05). Conclusions:Patients with high perianal abscess and treated with small incision floating line drainage has a shorter wound healing cycle, less pain, lower anal damage, and better clinical efficacy.

Ischemic stroke is a major public health problem worldwide. Although the circadian clock is involved in the process of ischemic stroke, the exact mechanism of the circadian clock in regulating angiogenesis after cerebral infarction remains unclear. In the present study, we determined that environmental circadian disruption (ECD) increased the stroke severity and impaired angiogenesis in the rat middle cerebral artery occlusion model, by measuring the infarct volume, neurological tests, and angiogenesis-related protein. We further report that Bmal1 plays an irreplaceable role in angiogenesis. Overexpression of Bmal1 promoted tube-forming, migration, and wound healing, and upregulated the vascular endothelial growth factor (VEGF) and Notch pathway protein levels. This promoting effect was reversed by the Notch pathway inhibitor DAPT, according to the results of angiogenesis capacity and VEGF pathway protein level. In conclusion, our study reveals the intervention of ECD in angiogenesis in ischemic stroke and further identifies the exact mechanism by which Bmal1 regulates angiogenesis through the VEGF-Notch1 pathway.
Rats ; Animals ; Vascular Endothelial Growth Factor A/pharmacology* ; Brain Ischemia/metabolism* ; Ischemic Stroke ; Signal Transduction ; ARNTL Transcription Factors/pharmacology* ; Neovascularization, Physiologic/physiology*

ObjectiveTo confirm the clinical efficacy and safety of Yishen Yangxin Anshen tablets in the treatment of insomnia （heart-blood deficiency and kidney-essence insufficiency syndrome）. MethodA randomized block， double-blind， placebo-controlled， multi-center clinical trial design method was adopted， and a total of 480 patients with insomnia due to deficiency of heart blood and insufficiency of kidney essence （treatment group-control group 3∶1） from seven hospitals （Guang'anmen Hospital， China Academy of Chinese Medical Sciences， The First Clinical Hospital， Jilin Province Academy of Traditional Chinese Medicine（TCM）， The Second Affiliated Hospital of Liaoning University of TCM， The First Affiliated Hospital of Henan University of Chinese Medicine， Henan Province Hospital of TCM， Hebei General Hospital， The First Hospital of Hunan University of Chinese Medicine） were enrolled. The treatment group was given Yishen Yangxin Anshen tablets and the control group received placebo tablets （4 tablets/time， 3 times/day， 4 weeks of administration， 4 weeks of follow-up after drug withdrawal）. The sleep dysfunction rating scale （SDRS） score， pittsburgh sleep quality index （PSQI） score， TCM， polysomnography （PSG） indicators from four hospital （Guang'anmen Hospital， China Academy of Chinese Medical Sciences， Henan Province Hospital of TCM， Hebei General Hospital， The First Hospital of Hunan University of Chinese Medicine）， and other efficacy indicators were compared between the two groups before and after treatment. Through general physical examination， laboratory examination， and observation of adverse events， the safety of the drugs was evaluated. ResultThe baseline indexes of the two groups showed no significant difference and thus the two groups were comparable. After treatment， the total score of SDRS in the treatment group was lower than that in the control group （P<0.01）. After drug withdrawal for 4 weeks， the total score of SDRS demonstrated no significant change in the treatment group as compared with that at the end of treatment， indicating that the rebound change of curative effect was not obvious. After treatment， the total score of PSQI in the treatment group decreased as compared with that in the control group （P<0.01）， and the change of total score of PSQI in the treatment group was statistically significant （P<0.05） after drug withdrawal for 4 weeks but small， indicating that the rebound change of curative effect was not obvious. After treatment， the total effective rate about the TCM symptoms in the treatment group was higher than that in the control group （χ²=137.521，P<0.01）. After treatment， the disappearance rates of single indexes in the treatment group， such as difficulty in falling asleep， easily waking up after sleeping， early awakening， short sleep time， dreamfulness， palpitation， forgetfulness， dizziness， mental fatigue， and weakness of waist and knee， increased compared with those in the control group （P<0.01）. After treatment， the treatment group demonstrated fewer awaking times （AT）， longer total sleep time （TST）， lower ATA/TST ratio， and higher sleep efficiency （%） than the control group （P<0.05）. No abnormal value or aggravation related to drugs was observed in either group. The incidence of adverse events in the treatment group and the control group was 5.57% and 8.40% respectively. No serious adverse events or adverse events leading to withdrawal happened in either group. ConclusionYishen Yangxin Anshen tablets is effective and safe for patients with insomnia of deficiency of heart-blood and insufficiency of kidney-essence.

To explore the pharmacogenomic markers that affect the platinum-based chemotherapy response in non-small-cell lung carcinoma (NSCLC), we performed a two-cohort of genome-wide association studies (GWAS), including 34 for WES-based and 433 for microarray-based analyses, as well as two independent validation cohorts. After integrating the results of two studies, the genetic variations related to the platinum-based chemotherapy response were further determined by fine-mapping in 838 samples, and their potential functional impact were investigated by eQTL analysis and in vitro cell experiments. We found that a total of 68 variations were significant at P < 1 × 10^-3 in cohort 1 discovery stage, of which 3 SNPs were verified in 262 independent samples. A total of 541 SNPs were significant at P < 1 × 10^-4 in cohort 2 discovery stage, of which 8 SNPs were verified in 347 independent samples. Comparing the validated SNPs in two GWAS, ADCY1 gene was verified in both independent studies. The results of fine-mapping showed that the G allele carriers of ADCY1 rs2280496 and C allele carriers of rs189178649 were more likely to be resistant to platinum-based chemotherapy. In conclusion, our study found that rs2280496 and rs189178649 in ADCY1 gene were associated the sensitivity of platinum-based chemotherapy in NSCLC patients.