[This corrects the article DOI: 10.1016/j.apsb.2022.03.002.].

BACKGROUND: Preclinical studies have indicated that Angong Niuhuang Pills (ANP) reduce cerebral infarct and edema volumes. This study aimed to investigate whether ANP safely reduces cerebral infarct and edema volumes in patients with moderate to severe acute ischemic stroke. METHODS: This randomized, double-blind, placebo-controlled pilot trial included patients with acute ischemic stroke with National Institutes of Health Stroke Scale (NIHSS) scores ranging from 10 to 20 in 17 centers in China between April 2021 and July 2022. Patients were allocated within 36 h after onset via block randomization to receive ANP or placebo (3 g/day for 5 days). The primary outcomes were changes in cerebral infarct and edema volumes after 14 days of treatment. The primary safety outcome was severe adverse events (SAEs) for 90 days. RESULTS: There were 57 and 60 patients finally included in the ANP and placebo groups, respectively for modified intention-to-treat analysis. The median age was 66.0 years, and the median NIHSS score at baseline was 12.0. The changes in cerebral infarct volume at day 14 were 0.3 mL and 0.4 mL in the ANP and placebo groups, respectively (median difference: -7.1 mL; interquartile range [IQR]: -18.3 to 2.3 mL, P = 0.30). The changes in cerebral edema volume of the ANP and placebo groups on day 14 were 11.4 mL and 4.0 mL, respectively ( median difference: 3.0 mL, IQR: -1.3 to 9.9 mL, P = 0.15). The rates of SAE within 90 days were similar in the ANP (3/57, 5%) and placebo (7/60, 12%) groups ( P = 0.36). Changes in serum mercury and arsenic concentrations were comparable. In patients with large artery atherosclerosis, ANP reduced the cerebral infarct volume at 14 days (median difference: -12.3 mL; IQR: -27.7 to -0.3 mL, P = 0.03). CONCLUSIONS: ANP showed a similar safety profile to placebo and non-significant tendency to reduce cerebral infarct volume in patients with moderate-to-severe stroke. Further studies are warranted to assess the efficacy of ANP in reducing cerebral infarcts and improving clinical prognosis. TRAIL REGISTRATION Clinicaltrials.gov , No. NCT04475328.
Aged ; Female ; Humans ; Male ; Middle Aged ; Double-Blind Method ; Drugs, Chinese Herbal/adverse effects* ; Ischemic Stroke/drug therapy* ; Pilot Projects ; Stroke/drug therapy* ; Treatment Outcome

Acute cerebral infarction predominantly occurs in middle-aged and elderly individuals with multiple underlying diseases， and it is often accompanied by atherosclerosis or heart diseases. It has a low incidence rate in children and adolescents， who tend to have atypical symptoms in the early stage， leading to misdiagnosis or missed diagnosis. With the continuous development of imaging technology， the diagnosis of cerebral infarction has become more precise， which helps to further clarify the etiology of cerebral infarction and identify an increasing number of patients with different subtypes of cerebral infarction in clinical practice. This article reports a rare case of a pediatric patient with acute cerebral infarction caused by primary central nervous system vasculitis， which led to neurological dysfunction， in order to improve the awareness of cerebral infarction in children among clinicians.
Vasculitis

Objective: To investigate the effects of Erianin on cell proliferation and apoptosis in human oral squamous cell carcinoma (OSCC) cells, providing a research foundation for the clinical treatment of OSCC. Methods: Erianin was applied to OSCC cells (CAL27 and SCC9) at concentrations of 0, 2.5, 5, and 10 μmol/L. The inhibitory effect of Erianin on OSCC cell proliferation was evaluated using CCK-8 and soft agar colony formation assays. Western blotting (WB) was employed to analyze the expression levels of anti-apoptotic proteins B-cell lymphoma-extra large (Bcl-xL), B-cell lymphoma-2 (Bcl-2), myeloid cell leukemia-1 (Mcl-1), and apoptotic protein cleaved-Caspase 3 (c-Caspase 3) in OSCC cells. Caspase 3 activity was further assessed using a caspase 3 activity detection kit to examine the pro-apoptotic effect of Erianin in OSCC cells. Mcl-1 overexpression was induced in CAL27 cells via plasmid transfection, and the influence of Mcl-1 on the effects of Erianin in CAL27 cells was analyzed by WB and caspase 3 activity measurement. All animal experiments were approved by the Ethics Committee of Hunan Cancer Hospital. A CAL27 xenograft mouse model was established and randomly divided into two groups (n = 5): the treatment group received intraperitoneal injection of Erianin (25 mg/kg), while the control group was injected with phosphate-buffered saline (PBS) as the vehicle. Immunohistochemistry (IHC) was used to detect the expression levels of Ki67 and Mcl-1 in the tumor tissues. Results: Erianin inhibited the proliferation of CAL27 and SCC9 cells in a dose-dependent manner and downregulated the protein expression of Mcl-1, with minimal effects on Bcl-2 and Bcl-xL. Furthermore, Erianin induced apoptosis in OSCC cells, as evidenced by increased expression of c-Caspase 3 and enhanced caspase 3 activity (P<0.001). Overexpression of Mcl-1 inhibited the Erianin-induced increase in c-Caspase 3 protein levels and caspase 3 activity. In vivo results were consistent with the in vitro findings. After Erianin treatment, CAL27 cell growth in nude mice was suppressed (P<0.001), and the expression levels of the proliferation marker Ki67 and the anti-apoptotic protein Mcl-1 in the tumor tissues were downregulated (P<0.001). Conclusion Erianin exhibits potent anti-tumor effects, effectively inhibiting the proliferation of OSCC cells and inducing apoptosis. The underlying mechanism may involve the downregulation of the pro-survival protein Mcl-1.

【Objective】 To identify the risk factors for recurrence and malignant transformation (MT) in patients with low-grade glioma (LGG) after surgery. 【Methods】 The data of 163 patients who underwent LGG resection and subsequent follow-up from March 2009 to April 2019 were retrospectively collected. Patients who did not experience recurrence or MT after surgery were included in the control group (85 cases), those who experienced recurrence after surgery were included in the observation 1 group (44 cases), and those who experienced MT after surgery were included in the observation 2 group (34 cases). Based on the clinical data of the three groups of patients, their clinical characteristics were analyzed, and the risk factors and predictive value for recurrence and MT were explored using Logistic regression model and receiver operating characteristic (ROC) curve. 【Results】 There were significant differences between the control group and the observation 1 group in preoperative seizure, preoperative Karnofsky performance status (KPS) score, and surgical approach (P<0.05). There were significant differences between the control group and the observation 2 group in gender, preoperative KPS score, tumor size, and surgical approach (P<0.05). There were significant differences between the control group and the observation 1 group in isocitrate dehydrogenase (IDH) mutation, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-9 (MMP-9), cancer-testis antigen OY-TES-1, OY-TES-1 mRNA protein, tumor suppressor protein p53, mouse double minute 2 (MDM2), vascular endothelial growth factor (VEGF), or epidermal growth factor receptor (EGFR) (P<0.05). There were significant differences between the control group and the observation 2 group in PCNA, MMP-9, cancer-testis antigen OY-TES-1, OY-TES-1 mRNA protein, or VEGF (P<0.05). Logistic regression analysis showed that IDH mutation, MMP-9, and PCNA were independent risk factors for LGG recurrence (P<0.05), while VEGF, MMP-9, and PCNA were independent risk factors for LGG MT (P<0.05). The area under curve (AUC) of PCNA, MMP-9 and IDH mutation for predicting LGG MT after surgery was 0.744, 0.790, and 0.799, respectively. The AUC of PCNA, MMP-9, and VEGF for predicting LGG recurrence after surgery was 0.729, 0.750, and 0.900, respectively. 【Conclusion】 This study found that IDH mutation, MMP-9 and PCNA were independent risk factors for LGG recurrence, while VEGF, MMP-9 and PCNA were independent risk factors for LGG MT by retrospectively analyzing the clinical data and protein expression of 163 patients with LGG after surgery. These proteins have high accuracy in predicting LGG recurrence and MT after surgery. Therefore, the proteins may play an important role in the biological behavior and treatment effect of LGG, and can be used as reference indicators for prognosis evaluation and individualized treatment of LGG patients after surgery.

Objective To investigate ecological isolation between Oncomelania hupensis snail populations in hilly regions and marshland and lake regions in Yuanjiang valley, Changde City, Hunan Province, and to unravel its underlying mechanisms. Methods Taoyuan County, Shimen County, Linli County and Lixian County in Changde City were selected as snail sampling sites in hilly regions, and Lixian County, Jinshi City, West Lake Administration District, Hanshou County and Dingcheng District were selected as snail sampling sites in marshland and lake areas. Cytochrome C oxidase 1 (cox 1) gene was amplified in snail samples and sequenced. The genetic sequences of O. hupensis snails were aligned using the software MEGA 11, and the haplotypes of O. hupensis snails were determined using the software DNASP 5.10.01. The phylogenetic tree was generated using Bayesian inference with the software MrBayes 3.2, and analysis of molecular variance (AMOVA) was performed to analyze the source of genetic divergence and estimate the genetic divergence index (F_ST) among snail populations with the software Arlequin 3.5.2.2. The genetic barrier among 11 O. hupensis snail populations was estimated using the Monmonier algorithm of adegenet toolkit in R package. The settings with “land in winter and water in summer” in the Yuanjian River section were divided into two categories according to the upstream and downstream, and the areas with “land in winter and water in summer” in the upstream and downstream were transformed into raster data, and then loaded into the software Fragstats 4 for analysis of landscape indicators. The trends in changes of digital elevation were extracted from the Yuanjiang River section based on the digital elevation model, and made three-dimensional visualization using the R package. Results The mitochondrial cox 1 gene were amplified in 165 O. hupensis snais from 11 sampling sites and sequenced, and a total of 152 valid gene sequences were obtained, with 46 haplotypes or 9 populations determined. No haplotype was shared in snails between Taoyuan County and Dingcheng District and Hanshou County along the downstream of the Yuanjiang River. The total area of settings with “land in winter and water in summer” was 617.66 hm² in the upsteram of the Yuanjiang River, which consisted of 473 patches, with each patch measuring 1.31 hm², the largest area index of 0.735 2, the landscape division index of 0.999 9, and the landscape shape index of 45.293 7. The total area of settings with “land in winter and water in summer” was 9 956.92 hm² in the downstream of the Yuanjiang River, which consisted of 771 patches, with each patch measuring 12.91 hm², the largest area index of 97.839 9, the landscape division index of 0.042 7, and the landscape shape index of 7.249 6. The area of settings with “land in winter and water in summer” was much larger in the downstream than that in the upstream of the Yuanjiang River, and the stronger landscape connectivity and non-remarkable alteration of riverbed elevation provided suitable habitats for snail breeding. Conclusion The hydrological and environmental characteristics of the upstream of the Yuanjiang River restrain the breeding and spread of O. hupensis, resulting in ecological isolation between Oncomelania hupensis in Taoyuan County and those in the downstream of Yuanjiang River.

Objective:The three-dimensional(3D) reconstruction method of calf perforator vessel was explored to provide the basis for clinical safe and accurate harvesting of calf chain perforator flap.Methods:Gelatin-lead oxide is automatically perfused the femoral artery of 3 fresh adult lower limbs specimens. Data were collected after the CT scan of the full limb, 3D reconstruction of bones, arteries and their perforators was processed in Mimics Research 19.0 software. Dynamic display of vascular hierarchy and adjacent relationships by adjusting CT values. And show the anatomical region of each perforator artery in different colors, measure the feeding area of the single perforator using Scion Image software. The 3D printed calf bone and vascular model can be used for direct observation. The specimen anatomy was measured according to a perforator vessel parameter with an outer diameter of 0.5 mm through the deep fascia. The number of perforators, vessle pedicle length, and outer diameter were observed and statistically analyzed.Results:The digitized 3D reconstruction images can clearly show the 3D structured of the calf artery and perforator vessels and micro vessels, as well as the course, distribution characteristics of blood vessels, and the 3D relationship with adjacent structures. The printed 3D model visually observes the hierarchy of the perforator vessels and the anastomosis relationship between the perforators. 3 calf specimen arteries with perforator artery 22±4, vessel pedicle length (46.7±18.3) mm, outer diameter (0.8±0.3) mm, the average blood supply area of the single perforator (38.2±10.7) cm 2. Conclusion:The Mimics Research 19.0 software should be used to dynamically reconstruct the 3D structure of the branch vessel by adjusting the CT value, which improves the display effect of the perforator vessels. The 3D printing model makes the source, course, diameter and distribution range of the perforator vessels visually visible, which provides experimental support for the clinical construction of digital calf chain perforator flap harvest scheme for patients.

OBJECTIVE: This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province. METHODS: We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed. RESULTS: The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α ^3.7/αα (50.23%) and β ^IVS-II-654/β ^N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively. CONCLUSION Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.
Humans ; beta-Thalassemia/genetics* ; alpha-Thalassemia/genetics* ; Hemoglobinopathies/genetics* ; China/epidemiology* ; High-Throughput Nucleotide Sequencing

Objective:The three-dimensional(3D) reconstruction method of calf perforator vessel was explored to provide the basis for clinical safe and accurate harvesting of calf chain perforator flap.Methods:Gelatin-lead oxide is automatically perfused the femoral artery of 3 fresh adult lower limbs specimens. Data were collected after the CT scan of the full limb, 3D reconstruction of bones, arteries and their perforators was processed in Mimics Research 19.0 software. Dynamic display of vascular hierarchy and adjacent relationships by adjusting CT values. And show the anatomical region of each perforator artery in different colors, measure the feeding area of the single perforator using Scion Image software. The 3D printed calf bone and vascular model can be used for direct observation. The specimen anatomy was measured according to a perforator vessel parameter with an outer diameter of 0.5 mm through the deep fascia. The number of perforators, vessle pedicle length, and outer diameter were observed and statistically analyzed.Results:The digitized 3D reconstruction images can clearly show the 3D structured of the calf artery and perforator vessels and micro vessels, as well as the course, distribution characteristics of blood vessels, and the 3D relationship with adjacent structures. The printed 3D model visually observes the hierarchy of the perforator vessels and the anastomosis relationship between the perforators. 3 calf specimen arteries with perforator artery 22±4, vessel pedicle length (46.7±18.3) mm, outer diameter (0.8±0.3) mm, the average blood supply area of the single perforator (38.2±10.7) cm 2. Conclusion:The Mimics Research 19.0 software should be used to dynamically reconstruct the 3D structure of the branch vessel by adjusting the CT value, which improves the display effect of the perforator vessels. The 3D printing model makes the source, course, diameter and distribution range of the perforator vessels visually visible, which provides experimental support for the clinical construction of digital calf chain perforator flap harvest scheme for patients.

Objective: The investigate the effects and mechanism of exosomes derived from human umbilical vein endothelial cells (HUVECs) on wound healing in diabetes rabbits. Methods: The experimental research methods were used. The primary vascular endothelial cells (VECs) and human skin fibroblasts (HSFs) were extracted from skin tissue around ulcer by surgical excision of two patients with diabetic ulcer (the male aged 49 years and the female aged 58 years) admitted to Xiangya Third Hospital of Central South University in June 2019. The cells were successfully identified through morphological observation and flow cytometry. The HUVEC exosomes were extracted by ultracentrifugation and identified successfully by morphological observation, particle size detection, and Western blotting detection. Twenty female 3-month-old New Zealand rabbits were taken to create one type 2 diabetic full-thickness skin defect wound respectively on both sides of the back. The wounds were divided into exosomes group and phosphate buffer solution (PBS) group and treated accordingly, with 20 wounds in each group, the time of complete tissue coverage of wound was recorded. On PID 14, hematoxylin-eosin staining or Masson staining was performed to observe angiogenesis or collagen fiber hyperplasia (n=20). The VECs and HSFs were co-cultured with HUVEC exosomes for 24 h to observe the uptake of HUVEC exosomes by the two kinds of cells. The VECs and HSFs were divided to exosome group treated with HUVEC exosomes and PBS group treated with PBS to detect the cell proliferation on 4 d of culture with cell count kit 8, to detect and calculate the cell migration rate at 24 and 48 h after scratch by scratch test, to detect the cell migration number at 24 h of culture with Transwell test, and to detect the mRNA expressions of nuclear factor-erythroid 2-related factor 2 (NRF2) and transcription activating factor 3 (ATF3) by real time fluorescence quantitative reverse transcription polymerase chain reaction. Besides, the number of vascular branches and vascular length were observed in the tube forming experiment after 12 h of culture of VECs (n=3). The VECs and HSFs were taken and divided into PBS group and exosome group treated as before, and NRF2 interference group, ATF3 interference group, and no-load interference group with corresponding gene interference. The proliferation and migration of the two kinds of cells, and angiogenesis of VECs were detected as before (n=3). Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, independent sample t test, and least significant difference test. Results: The time of complete tissue coverage of wound in exosome group was (17.9±1.9) d, which was significantly shorter than (25.2±2.3) d in PBS group (t=4.54, P<0.05). On PID14, the vascular density of wound in PBS group was significantly lower than that in exosome group (t=10.12, P<0.01), and the collagen fiber hyperplasia was less than that in exosome group. After 24 h of culture, HUVEC exosomes were successfully absorbed by VECs and HSFs. The proliferative activity of HSFs and VECs in exosome group was significantly higher than that in PBS group after 4 d of culture (with t values of 54.73 and 7.05, respectively, P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs (with t values of 3.42 and 11.87, respectively, P<0.05 or P<0.01) and VECs (with t values of 21.42 and 5.49, respectively, P<0.05 or P<0.01) in exosome group were significantly higher than those in PBS group. After 24 h of culture, the migration numbers of VECs and HSFs in exosome group were significantly higher than those in PBS group (with t values of 12.31 and 16.78, respectively, P<0.01). After 12 h of culture, the mRNA expressions of NRF2 in HSFs and VECs in exosome group were significantly higher than those in PBS group (with t values of 7.52 and 5.78, respectively, P<0.05 or P<0.01), and the mRNA expressions of ATF3 were significantly lower than those in PBS group (with t values of 13.44 and 8.99, respectively, P<0.01). After 12 h of culture, the number of vascular branches of VECs in exosome group was significantly more than that in PBS group (t=17.60, P<0.01), and the vascular length was significantly longer than that in PBS group (t=77.30, P<0.01). After 4 d of culture, the proliferation activity of HSFs and VECs in NRF2 interference group was significantly lower than that in PBS group and exosome group (P<0.05 or P<0.01); the proliferation activity of HSFs and VECs in ATF3 interference group was significantly higher than that in PBS group (P<0.05 or P<0.01) and significantly lower than that in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in ATF3 interference group were significantly higher than those in PBS group (P<0.05 or P<0.01) and significantly lower than those in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in NRF2 interference group were significantly lower than those in PBS group and exosome group (P<0.05 or P<0.01). After 24 h of culture, the migration numbers of VECs and HSFs in ATF3 interference group were significantly more than those in PBS group (P<0.05) and significantly less than those in exosome group (P<0.05 or P<0.01); the migration numbers of VECs and HSFs in NRF2 interference group were significantly less than those in PBS group and exosome group (P<0.01). After 12 h of culture, the vascular length and number of branches of VECs in NRF2 interference group were significantly decreased compared with those in PBS group and exosome group (P<0.01); the vascular length and number of branches of VECs in ATF3 interference group were significantly increased compared with those in PBS group (P<0.01) and were significantly decreased compared with those in exosome group (P<0.01). Conclusions: HUVEC exosomes can promote the wound healing of diabetic rabbits by promoting the proliferation and migration of VECs and HSFs, and NRF2 and ATF3 are obviously affected by exosomes in this process, which are the possible targets of exosome action.
Animals ; Female ; Humans ; Male ; Rabbits ; Collagen/metabolism* ; Diabetes Mellitus ; Exosomes/metabolism* ; Human Umbilical Vein Endothelial Cells ; Hyperplasia/metabolism* ; NF-E2-Related Factor 2/metabolism* ; RNA, Messenger/metabolism* ; Ulcer ; Wound Healing ; Middle Aged