Analysis of clinicopathological and molecular abnormalities of angioimmunoblastic T-cell lymphoma / 北京大学学报(医学版)

OBJECTIVE@#To analyze the clinicopathological features, molecular changes and prognostic factors in angioimmunoblastic T-cell lymphoma (AITL).@*METHODS@#Sixty-one cases AITL diagnosed by Department of Pathology of Peking University Cancer Hospital were collected with their clinical data. Morphologically, they were classified as typeⅠ[lymphoid tissue reactive hyperplasia (LRH) like]; typeⅡ[marginal zone lymphoma(MZL)like] and type Ⅲ [peripheral T-cell lymphoma, not specified (PTCL-NOS) like]. Immunohistochemical staining was used to evaluate the presence of follicular helper T-cell (TFH) phenotype, proliferation of extra germinal center (GC) follicular dendritic cells (FDCs), presence of Hodgkin and Reed-Sternberg (HRS)-like cells and large B transformation. The density of Epstein-Barr virus (EBV) + cells was counted with slides stained by Epstein-Barr virus encoded RNA (EBER) in situ hybridization on high power field (HPF). T-cell receptor / immunoglobulin gene (TCR/IG) clonality and targeted exome sequencing (TES) test were performed when necessary. SPSS 22.0 software was used for statistical analysis.@*RESULTS@#Morphological subtype (%): 11.4% (7/61) cases were classified as type Ⅰ; 50.8% (31/61) as type Ⅱ; 37.8% (23/61) as type Ⅲ. 83.6% (51/61) cases showed classical TFH immunophenotype. With variable extra-GC FDC meshwork proliferation (median 20.0%); 23.0% (14/61) had HRS-like cells; 11.5% (7/61) with large B transformation. 42.6% (26/61) of cases with high counts of EBV. 57.9% (11/19) TCR+/IG-, 26.3% (5/19) TCR+/IG+, 10.5% (2/19) were TCR－/IG－, and 5.3% (1/19) TCR－/IG+. Mutation frequencies by TES were 66.7% (20/30) for RHOA, 23.3% (7/30) for IDH2 mutation, 80.0% (24/30) for TET2 mutation, and 33.3% (10/30) DNMT3A mutation. Integrated analysis divided into four groups: (1) IDH2 and RHOA co-mutation group (7 cases): 6 cases were type Ⅱ, 1 case was type Ⅲ; all with typical TFH phenotype; HRS-like cells and large B transformation were not found; (2) RHOA single mutation group (13 cases): 1 case was type Ⅰ, 6 cases were type Ⅱ, 6 cases were type Ⅲ; 5 cases without typical TFH phenotype; 6 cases had HRS-like cells, and 2 cases with large B transformation. Atypically, 1 case showed TCR－/IG－, 1 case with TCR－/IG+, and 1 case with TCR+/IG+; (3) TET2 and/or DNMT3A mutation alone group (7 cases): 3 cases were type Ⅱ, 4 cases were type Ⅲ, all cases were found with typical TFH phenotype; 2 cases had HRS-like cells, 2 cases with large B transformation, and atypically; (4) non-mutation group (3 cases), all were type Ⅱ, with typical TFH phenotype, with significant extra-GC FDC proliferation, without HRS-like cells and large B transformation. Atypically, 1 case was TCR－/IG－. Univariate analysis confirmed that higher density of EBV positive cell was independent adverse prognostic factors for both overall survival (OS) and progression free survival(PFS), (P=0.017 and P=0.046).@*CONCLUSION@#Pathological diagnoses of ALTL cases with HRS-like cells, large B transformation or type Ⅰ are difficult. Although TCR/IG gene rearrangement test is helpful but still with limitation. TES involving RHOA, IDH2, TET2, DNMT3A can robustly assist in the differential diagnosis of those difficult cases. Higher density of EBV positive cells counts in tumor tissue might be an indicator for poor survival.

Clinicopathological features and prognosis of anorectal melanoma: A report of 68 cases / 北京大学学报(医学版)

OBJECTIVE@#To investigate the clinicopathological characteristics of anorectal mucosal melanoma (ARMM), and to evaluate the prognostic factors.@*METHODS@#A total of 68 primary ARMM surgical specimens from 2010 to 2018 were retrospectively studied. Slides were reviewed to evaluate pathological features. Slingluff staging method was used for staging.@*RESULTS@#(1) Clinical features: The median age at diagnosis in this group was 61.5 years, with a male-to-female ratio 1 ∶1.62. The most common complaint was blooding (49 cases). For anatomic site, anorectum was the prevalent (66.2%), followed by rectum (20.6%). At the time of diagnosis, 28 cases were stage Ⅰ (localized stage, 41.2%), 25 cases were stage Ⅱ (regional lymph node metastasis, 36.8%), and 15 cases were stage Ⅲ (distant metastasis, 22.1%). Five patients underwent wide local excision, the rest abdominoperineal resection, and 48 patients received adjuvant therapy after surgery. (2) Pathological features: Grossly 88.2% of the tumors were exophytic polypoid masses, with the median tumor size 3.5 cm and the median tumor thickness 1.25 cm. Depth of invasion below lamina muscularis mucosae ranged from 0-5.00 cm (median 1.00 cm). The deepest site of tumor invasion reached muscular layer in 27 cases, and perirectal tissue in 16 cases. Melanin pigmentation was absent or not obvious in 67.6% of the cases. The predominant cytology was epithelioid (45 cases, 66.2%). The rate for ulceration, necrosis, lymphovascular invasion, and perineural invasion was 89.7%, 35.3%, 55.9%, and 30.9%, respectively. The median mitotic count was 18/mm2. The positive rate of S100, HMB-45 and Melan-A were 92.0%, 92.6% and 98.0%, respectively. The median of Ki-67 was 50%. The incidences of mutations within CKIT, BRAF and NRAS genes were 17.0% (9 cases), 3.8% (2 cases) and 9.4% (5 cases), respectively. (3) Prognosis: Survival data were available in 66 patients, with a median follow-up of 17 months and a median survival time of 17.4 months. The 1-year, 2-year and 5-year overall survival rate was 76.8%, 36.8% and 17.2%, respectively. The rate of lymphatic metastasis at diagnosis was 56.3%. Forty-nine patients (84.5%) suffered from distant metastasis, and the most frequent metastatic site was liver. Univariate analysis revealed that tumor size (>3.5 cm), depth of invasion below lamina muscularis mucosae (>1.0 cm), necrosis, lymphovascular invasion, BRAF gene mutation, lack of adjuvant therapy after surgery, deep site of tumor invasion, and high stage at diagnosis were all poor prognostic factors for overall survival. Multivariate model showed that lymphovascular invasion and BRAF gene mutation were independent risk factors for lower overall survival, and high stage at diagnosis showed borderline negative correlation with overall survival.@*CONCLUSION@#The overall prognosis of ARMM is poor, and lymphovascular invasion and BRAF gene mutation are independent factors of poor prognosis. Slingluff staging suggests prognosis effectively, and detailed assessment of pathological features, clear staging and genetic testing should be carried out when possible. Depth of invasion below lamina muscularis mucosae of the tumor might be a better prognostic indicator than tumor thickness.

Measurement of the Thermic Effect of Food in a Chinese Mixed Diet in Young People / 生物医学与环境科学（英文）

OBJECTIVE@#To determine the thermic effect of food (TEF) in a Chinese mixed diet in young people.@*METHODS@#During the study, the participants were weighed and examined for body composition every morning. The total energy expenditure (TEE) of the participants was measured by the doubly labeled water method for 7 days, and during this period, basal energy expenditure was measured by indirect calorimetry and physical activity energy expenditure was measured by an accelerometer. The value obtained by subtracting basal energy expenditure and physical activity energy expenditure from TEE was used to calculate TEF.@*RESULTS@#Twenty healthy young students (18-30 years; 10 male) participated in the study. The energy intake of the participants was not significantly different from the Chinese Dietary Reference Intake of energy ( P > 0.05). The percentage of energy from protein, fat and carbohydrate were all in the normal range. The intakes of fruits, milk and dietary fiber of the participants were significantly lower than those in the Chinese Dietary Guidelines ( P < 0.05). There was no significant difference in the body weight of the participants during the experiment ( P > 0.05). When adjusted for body weight, there was no significant difference in either TEE or basal energy expenditure between the male and female participants ( P > 0.05). In addition, there was no significant difference in physical activity energy expenditure and TEF between the male and female participants ( P > 0.05). The percentage of TEF in TEE was 8.73%.@*CONCLUSION@#The percentage of TEF in TEE in a Chinese mixed diet in young people was significantly lower than 10% ( P < 0.001). A value of 10% is usually considered to be the TEF in mixed diets as a percentage of TEE.

Effect of sodium bicarbonate Ringer′s solution on acute kidney injury following laparoscopic hepatectomy in elderly patients / 中华麻醉学杂志

Objective:To evaluate the effect of sodium bicarbonate Ringer′s solution on acute kidney injury(AKI) following laparoscopic hepatectomy in elderly patients.Methods:A total of 362 American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ elderly patients, aged 65-79 yr, with body mass index of 18-28 kg/m 2, scheduled for elective laparoscopic hepatectomy, were divided into 2 groups( n=181 each) using a random number table method: bicarbonate Ringer′s solution group(BR group) and lactated Ringer′s solution group(LR group). Bicarbonate Ringer′s solution and lactated Ringer′s solution were intravenously infused in BR group and LR group, respectively. All operations were performed under general anesthesia combined with abdominal fascia block, and the methods of controlled low central venous pressure and intermittent hepatic inflow occlusion were applied to reduce intraoperative bleeding. Radial artery blood samples were collected for blood gas analysis at 5 min before anesthesia induction(T 0), 20 min after occluding liver hilus(T 1), 10 min after hepatectomy and hemostasis(T 2), at the end of surgery(T 3) and at postanesthesia care unit discharge(T 4), and lactate value(Lac) was recorded. Blood samples from cubital vein were collected on admission to hospital(T A) and at 24(T 24) and 48 h after operation(T 48) for determination of serum creatinine(Cr) concentrations. Doppler-based renal resistive index(RRI) was measured at T A, T 4, T 24 and T 48. The incidence of AKI was calculated within 48 h after operation according to the Kidney Disease: Improving Global Outcomes criteria in 2012 for Cr concentration. Adverse reactions(such as nausea and vomiting) and complications(such as incision infection) within 48 h after operation were recorded. Results:Compared with the baseline at T 0, Lac concentrations were significantly increased at T 1-4 in both groups( P<0.01). Cr concentrations were significantly increased at T 24 and T 48, and RRI was increased at T 4, T 24 and T 48 than at T A in both groups( P<0.01). Compared with group LR, the incidence of AKI within 48 h after operation, Lac concentrations at T 3, 4, Cr concentrations at T 24 and T 48, and RRI at T 4, T 24 and T 48 were significantly decreased in group BR( P<0.05 or 0.01). There was no significant difference in the incidence of nausea, vomiting, incision infection, delirium, bile leakage and pulmonary infection within 48 h after operation among the two groups( P>0.05). Conclusions:Sodium bicarbonate Ringer′s solution can decrease the development of AKI following laparoscopic hepatectomy in elderly patients.

Effect of selective cerebral mild hypothermia on SUMO2/3 modification of Drp1 in a rat model of cerebral ischemia-reperfusion / 中华麻醉学杂志

Objective:To evaluate the effect of selective cerebral mild hypothermia on small ubiquitin-like modifier 2/3 (SUMO2/3) modification of dynamin-related protein 1 (Drp1) in a rat model of cerebral ischemia-reperfusion (I/R).Methods:Sixty clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 240-260 g, were divided into 4 groups ( n=15 each) using a random number table method: sham operation group (S group), cerebral I/R group (I/R group), selective cerebral mild hypothermia group (HT group) and normal temperature group (NT group). The operation was performed under the monitoring of cerebral temperature and rectal temperature.Only the cervical blood vessels were exposed in S group, while focal cerebral I/R was induced by 2 h middle cerebral artery occlusion (MCAO) followed by 24 h reperfusion in anesthetized animals in the other three groups.In HT group and NT group, 4 and 37 ℃ normal saline was perfused through the left internal carotid artery at a rate of 80 ml·kg -1·h -1 for 15 min, respectively. Modified neurological severity score (mNSS) was assessed at 24 h of reperfusion. Then the rats were sacrificed under deep anesthesia, brains were removed, brain tissues were obtained for determination of the percentage of cerebral infarct size (by TTC staining), and the ischemic penumbra tissues in the cerebral cortex were removed for examination of the ultra-structural changes of mitochondria (with a transmission electron microscope) and for determination of the SUMO2/3 modification of Drp1 (by CO-IP), expression of total Drp1 (T-Drp1) and total cytochrome c (T-Cytc) (by Western blot), and expression of mitochondrial outer membrane Drp1 (M-Drp1) and cytoplasmic Cytc (C-Cytc) (by Western blot) after isolation of mitochondria and cytoplasm. Results:Compared with S group, the mNSS and percentage of cerebral infarct size were significantly increased, the expression of M-Drp1, T-Drp1, C-Cytc and T-Cytc was up-regulated, and SUMO2/3 modification of Drp1 in ischemic penumbra area was increased ( P<0.05), the fragmentation of mitochondria was aggravated, and cristae rupture and vacuolation were obvious in the other three groups. Compared with I/R group, the mNSS and percentage of cerebral infarct size were significantly decreased, the expression of M-Drp1, T-Drp1, C-Cytc and T-Cytc was down-regulated, SUMO2/3 modification of Drp1 was increased ( P<0.05), the fragmentation of mitochondria was significantly attenuated, and cristae rupture and vacuolation were weakened in HT group. There were no significant differences in these detection parameters between NT group and I/R group ( P>0.05). Conclusions:The mechanism by which selective cerebral mild hypothermia alleviates the cerebral I/R injury is related to increased SUMO2/3 modification of Drp1, decreased binding of Drp1 to mitochondrial outer membrane, and reduced mitochondrial excessive fission in rats.

Effects of mild hypothermia on microglia polarization and JAK2/STAT3 signaling pathway during cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective:To investigate the effects of mild hypothermia on microglia polarization and janus kinase 2/signal transduction and transcriptional activation factor 3 (JAK2/STAT3) signaling pathway during cerebral ischemia-reperfusion (I/R) in rats.Methods:Forty-five clean-grade healthy male Sprague-Dawley rats, aged 8 weeks, weighing 260-280 g, were divided into 3 groups ( n=15 each) by the random number table method: sham operation group (S group), cerebral I/R group (I/R) and mild hypothermia group (H group). In I/R group and H group, cerebral I/R was induced by middle cerebral artery occlusion using a nylon thread in anesthetized animals, the nylon thread was removed to restore the perfusion after 2 h of occlusion, and the rectal temperature was maintained at 36-37 ℃ during the period. Group H was wiped with 75% alcohol for 3 h starting from the time point immediately after reperfusion, and the rectal temperature was maintained at 32-33℃. Modified neurological severity score (mNSS) was evaluated at 24 h of reperfusion. Animals were then sacrificed for determination of the cerebral infarct size (using TTC staining), expression of M1 marker inducible nitric oxide synthase (iNOS), M2 marker arginase 1(Arg-1), phosphorylated JAK2(p-JAK2)and phosphorylated STAT3(p-STAT3)(by Western blot), expression of iNOS mRNA and Arg-1 mRNA (by quantitative polymerase chain reaction), and contents of interleukin-6 (IL-6) and IL-10 (by enzyme-linked immunosorbent assay). Results:Compared with group S, mNSS and cerebral infarct size were significantly increased, the expression of iNOS, Arg-1 protein and mRNA in cerebral ischemic penumbral zone was up-regulated, and the p-JAK2/JAK2 ratio, p-STAT3/STAT3 ratio, and contents of IL-6 and IL-10 were increased in the other two groups ( P<0.05). Compared with I/R group, mNSS and cerebral infarct size were significantly decreased, the expression of iNOS protein and mRNA in cerebral ischemic penumbral zone was down-regulated, the expression of Arg-1 and mRNA was up-regulated, and the p-JAK2/JAK2 ratio, p-STAT3/STAT3 ratio and IL-6 content were decreased, and the IL-10 content was increased in group H ( P<0.05). Conclusions:Mild hypothermia can promote the polarization shift of microglia from M1 to M2 phenotype during cerebral I/R and inhibit the central inflammatory responses, and the mechanism may be related to inhibition of JAK2/STAT3 signaling pathway in rats.

Relationship between hippocampal miR-3065-5p and IGF-1/PI3K/Akt signaling pathway in a mouse model of perioperative neurocognitive disorder / 中华麻醉学杂志

Objective:To evaluate the relationship between hippocampal miR-3065-5p and insulin-like growth factor-1/phosphatidylinositol 3-kinase/protein kinase B(IGF-1/PI3K/Akt)signaling pathway in a mouse model of perioperative neurocognitive disorder (PND).Methods:Eighty clean-grade healthy male C75BL/6 mice, aged 12-14 weeks, weighing 20-30 g, were divided them into 4 groups ( n=20 each) using the random number table method: control group (C group), PND group, miR-3065-5p agonist group (Ag group) and miR-3065-5p agonist negative control group (Ag-NC group). PND model was prepared by internal fixation of tibial fracture under anesthesia with 1.5% isoflurane. Two days before developing the model, miR-3065-5p agomir 2 μl was injected into the lateral ventricle in Ag group, miR-3065-5p agomir negative control 2 μl was injected into the lateral ventricle in Ag-NC group. Morris water maze test and open field test were performed at 7 days after surgery. The mice were sacrificed after the end of test, and hippocampal tissues were obtained for determination of the expression of miR-3065-5p, IGF-1 mRNA and Bcl-2 mRNA (by quantitative real-time polymerase chain reaction) and expression of IGF-1, phosphorylated Akt (p-Akt), phosphorylated glycogen synthase kinase-3β (p-GSK3β) and Bcl-2 (by Western blot). Results:There was no significant difference in each parameter in the open field test among the four groups ( P>0.05). Compared with group C, the postoperative escape latency was significantly prolonged, the percentage of time of stay at the target quadrant was decreased, the number of crossing the original platform was reduced, the expression of miR-3065-5p was up-regulated, and the expression of IGF-1 mRNA, Bcl-2 mRNA, IGF-1, p-Akt, p-GSK3β and Bcl-2 was down-regulated in the other three groups ( P<0.05). Compared with PND group and Ag-NC group, the postoperative escape latency was significantly prolonged, the percentage of time of stay at the target quadrant was decreased, the number of crossing the original platform was reduced, the expression of miR-3065-5p was up-regulated, and the expression of IGF-1 mRNA, Bcl-2 mRNA, IGF-1, p-Akt, p-GSK3β and Bcl-2 was down-regulated in Ag group ( P<0.05). Conclusions:Up-regulation of miR-3065-5p can inhibit the activation of IGF-1/PI3K/Akt signaling pathway, which might be one of the mechanisms of PND developed in mice.

Effects of remimazolam on gastrointestinal motor function in patients undergoing gastrointestinal endoscopy / 中华麻醉学杂志

Objective:To evaluate the effects of remimazolam on gastrointestinal motor function in the patients undergoing gastrointestinal endoscopy.Methods:A total of 262 American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-64 yr, with body mass index of 18-28 kg/m 2, scheduled for elective gastrointestinal endoscopy from May 2022 to August 2022, were divided into 2 groups ( n=131 each) using a random number table method: remimazolam group (group R) and propofol group (group P). The patients in group R received intravenous remimazolam 0.20-0.25 mg/kg, and patients in group P received intravenous propofol 1.5-2.0 mg/kg. The gastrointestinal endoscopy was performed when the patients′ Modified Observer′s Assessment of Alertness/Sedation scores ≤3. During fasting before gastrointestinal preparation, before gastrointestinal endoscopy and while leaving the post-anesthesia care unit (PACU), the concentrations of serum motilin and gastrin were measured by enzyme-linked immunosorbent assay, the intestinal peristalsis rating assessed by the endoscopist during the examination was recorded, the occurrence of hypotension and hypoxemia during the examination and occurrence of abdominal distension, abdominal pain, and nausea and vomiting during stay in PACU were recorded. Results:Compared with group P, the intestinal peristalsis rating was significantly increased, the serum motilin and gastrin concentrations were increased while leaving PACU, the incidence of hypotension and hypoxemia was decreased during the examination, and the incidence of abdominal distention was decreased during stay in PACU in group R ( P<0.05). Conclusions:Remimazolam has a milder inhibitory effect on secretion of gastrointestinal hormones than propofol in the patients undergoing gastrointestinal endoscopy and is helpful for the recovery of gastrointestinal motility.

Bibliometrics Analysis in English and Chinese Literature on Drowning in Forensic Medicine from 1991 to 2020 / 法医学杂志

OBJECTIVES@#To explore the research hotspots and development trends of the field of forensic drowning from 1991 to 2020 by bibliometrics methods.@*METHODS@#Based on Web of Science, CNKI database, Wanfang Data knowledge service platform, python 3.9.2, CiteSpace 5.8.R3, Gephi 0.9.2, etc. were used to analyze the publishing trends, countries/regions, institutions, authors and topics of the study on drowning.@*RESULTS@#A total of 631 English literature were obtained, including 59 articles from Chinese authors, and 386 Chinese literature were obtained. The Chinese and English journals with the largest number of related literatures were Chinese Journal of Forensic Science (80 articles) and Forensic Science International (106 articles), respectively. Japan published the most articles in English, and China ranked third. Osaka City Univ (Japan, 28 articles) published the most English articles, and Guangzhou Forens Sci Inst (China, 22 articles) ranked second. Among Chinese literature, Guangzhou Forens Sci Inst (32 articles) published the most. The topic analysis of Chinese and English literature showed that diatom examination, virtual autopsy, postmortem biochemical examination, the nature of death, and postmortem submersion interval were the hot spots of current research, but English literature had more studies on new technologies and methods, while Chinese literature was more inclined to practice, application and experience summary.@*CONCLUSIONS@#The number of literature in forensic medicine on drowning is relatively stable. The scope of international and domestic collaborations in this field is still limited. The automated examination of diatoms, the establishment of diatom DNA barcodes and virtual autopsy will be the most important research hotspots in the coming period and are expected to achieve breakthroughs in drowning diagnosis, drowning location inference, postmortem submersion interval estimation, etc.

Establishment of a Mouse Model of Acquired Aplastic Anemia Mediated by Cyclophosphamide Combined with Cyclosporine / 中国实验血液学杂志

OBJECTIVE@#To establish a stable mouse model of acquired aplastic anemia.@*METHODS@#Female BALB/C mice aged 6 months were intraperitoneally injected with cyclophosphamide and cyclosporine for 14 days. The number of peripheral blood cells, the concentration of hemoglobin, the number of bone marrow nucleated cells, bone marrow smear, bone marrow pathological sections and other indexes were observed.@*RESULTS@#In BALB/C mice injected intraperitoneally with cyclophosphamide and cyclosporine, the number of peripheral blood cells and the concentration of hemoglobin were significantly decreased, especially the white blood cells and platelets. Bone marrow smear showed a significant decrease in the number of nucleated cells and bone marrow hyperplasia. Bone marrow pathology showed decreased hematopoietic cells and increased non-hematopoietic cells such as adipocytes.@*CONCLUSION@#The mouse model with intraperitoneal injection of cyclophosphamide and cyclosporine can meet the diagnostic criteria of acquired aplastic anemia, which can be used as a mouse model for the study of the pathogenesis and treatment of acquired aplastic anemia.

Relationship between microRNA-93-5p and mitochondrial fusion protein-2 in mouse nerve cells subjected to oxygen-glucose deprivation and reoxygenation / 中华麻醉学杂志

Objective:To evaluate the relationship between microRNA-93-5p and mitochondrial fusion protein-2 (Mfn2) in mouse nerve cells subjected to oxygen-glucose deprivation and reoxygenation (OGD/R).Methods:Mouse neuroblastoma cells were cultured in vitro to logarithmic growth phase.Experiment Ⅰ Cells were divided into 5 groups ( n=20 each) by the random number table method: control group (group C), group OGD/R, miR-93-5p inhibitor group (group I), siRNA-mfn2 plus miR-93-5p group (group siMfn2+ I) and miR-93-5p negative control group (group NC). Oxygen-glucose deprivation: the cells were cultured for 3 h in a low-glucose balanced salt solution at 37 ℃ in an environment of 5% CO 2-95% N 2.Restoration of oxygen and glucose: the cells were cultured in normal medium at 37℃ in 5% CO 2-95% air for 24 h. Group I, group siMfn2+ I and group NC were transfected with miR-93-5p inhibitor, miR-93-5p inhibitor plus siRNA-mfn2 and negative control miRNA, respectively, at 48 h before the OGD/R model was developed.Cell viability was measured by CCK-8 assay.Cell apoptosis rate was measured by flow cytometry.Quantitative real-time polymerase chain reaction was used to detect the expression of miR-93-5p and Mfn2 mRNA.Western blot was used to detect Mfn2 protein expression.Experiment Ⅱ The wild-type (WT)-Mfn2 and mutant (MUT)-Mfn2 were constructed and transfected into neuroblastoma cells with miR-93-5p mimic and miR-93-5p blank control (miR-93-5pNC), respectively.The cells were divided into 4 groups ( n=5 each) after 48 h of transfection by the random number table method: miR-93-5p NC-WT-Mfn2 co-transfection group, miR-93-5p mimic-WT-Mfn2 co-transfection group, miR-93-5p NC-MUT-Mfn2 co-transfection group, and miR-93-5p mimic-MUT-Mfn2 co-transfection group.The activity of luciferases was measured by double luciferase assay. Results:Experiment Ⅰ Compared with group C, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of miR-93-5p was up-regulated, and the expression of Mfn2 protein and mRNA was down-regulated in the other gorups ( P<0.05). Compared with group OGD/R or group NC, the cell viability was significantly increased, the apoptosis rate of cells was decreased, miR-93-5p expression was down-regulated, and the expression of Mfn2 protein and mRNA was up-regulated in group I ( P<0.05). Compared with group I, the cell viability was significantly decreased, the apoptosis rate of cells was increased, and the expression of Mfn2 protein and mRNA was down-regulated in group siMfn2+ I ( P<0.05). Experiment Ⅱ Compared with miR-93-5p NC-WT-Mfn2 co-transfection group, the luciferase activity was significantly decreased in miR-93-5p mimic-WT-Mfn2 co-transfection group ( P<0.05). There was no significant difference in luciferase activity between miR-93-5p NC-MUT-Mfn2 co-transfection group and miR-93-5p mimic-MUT-Mfn2 co-transfection group ( P>0.05). Conclusions:The miR-93-5p expression is up-regulated, which further targetedly down-regulates the expression of Mfn2, and this may be a mechanism of OGD/R in mouse nerve cells.

Gastric emptying of orally administered enzyme-hydrolyzed rice flour solution before surgery in patients undergoing laparoscopic cholecystectomy and effect on insulin resistance / 中华麻醉学杂志

Objective:To evaluate the gastric emptying of orally administered enzyme-hydrolyzed rice flour solution before surgery in the patients undergoing laparoscopic cholecystectomy and effect on insulin resistance.Methods:One hundred patients, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, aged 18-64 yr, with body mass index of 19-30 kg/m 2, scheduled for elective laparoscopic cholecystectomy under general anesthesia, were divided into 2 groups ( n=50 each) using a random number table method: water group (group C) and enzyme-hydrolyzed rice flour group (group M). Routine fasting and water deprivation were executed at 1 day before operation in two groups, and 300 ml water in group C or 300 ml enzyme-hydrolyzed rice flour solution in group M were taken orally at 2-3 h before induction on the day of surgery.Bedside antrum ultrasonography was used to calculate the gastric volume (GV) before oral administration (V 0), immediately after oral administration (V 1), and before induction (V 2), and then the ΔGV (GV 1-GV 0) was calculated.Fasting plasma glucose and insulin CONCENTRATIONS were measured on admission to hospital (T 1) and on an empty stomach on 1st morning after surgery (T 2), and then the homeostasis model assessment of insulin resistance (HOMA-IR) was calculated according to HOMA steady-state model formula.Visual analog scale (VAS) scores for subjective comfort (thirst, hunger, fatigue and anxiety) and grip strength were assessed before anesthesia (T 3) and before leaving PACU (T 4). Reflux and aspiration during induction, nausea and vomiting within 24 h after surgery, and anal exhaust time after surgery were recorded. Results:There was no significant difference in GV at V 0, V 1 and V 2 between the two groups ( P>0.05). Compared with the baseline at V 0, no significant was found in the GV at V 2 in both groups ( P>0.05). The fasting plasma glucose and insulin concentrations and HOMA-IR were significantly increased at T 2 than at T 1 in both groups ( P<0.05 or 0.01). Compared with group C, the fasting plasma glucose and insulin concentrations and HOMA-IR were significantly decreased at T 2, VAS scores for hunger, fatigue and anxiety were decreased at T 3, 4, grip strength was increased at T 3, 4, the postoperative anal exhaust time was shortened, and the incidence of nausea was reduced in group M ( P<0.05). No reflux and aspiration happened during induction in either group. Conclusion:The gastric emptying of 300 ml enzyme-hydrolyzed rice flour solution orally administered at 2 h before surgery is normal in the patients undergoing laparoscopic cholecystectomy, which does not increase the risk of reflux and aspiration during anesthesia induction, reduces postoperative insulin resistance, and increases patient′s subjective comfort, and enhances the postoperative recovery of intestinal function.

Role of hsa_circ_0025853 in hypothermia-induced reduction of oxygen-glucose deprivation and restoration injury to neuroblastoma cells / 中华麻醉学杂志

Objective:To evaluate the role of hsa_circ_0025853 in hypothermia-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury to neuroblastoma cells (SK-N-SH cells). Methods:SK-N-SH cells were cultured in vitro to the logarithmic phase and divided into 4 groups ( n=20 each) using a random number table method: control group (group C), group OGD/R, hypothermia group (group H) and hsa_circ_0025853 small interfering RNA (siRNA) plus hypothermia group (group S+ H). The cells were cultured in normal culture atmosphere in group C. The cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply for 24 h in group OGD/R.The cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply for 24 h at 32 ℃ in group H. The cells were transfected with siRNA at 48 h before establishing OGD/R model, and the other treatments were similar to those previously described in group H. At 24 h of restoration of oxygen-glucose, cell viability was measured by Cell Counting Kit-8, apoptosis rate were measured by flow cytometry.the expression of dynamin-related protein 1 (Drp1) mRNA and hsa_circ_0025853 was detected by quantitative real-time-polymerase chain reaction, and the expression of Drp1 protein was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of hsa_circ_0025853 was down-regulated, and the expression of Drp1 protein and mRNA was up-regulated in the other 3 groups ( P<0.05). Compared with group OGD/R, the cell viability was significantly increased, the apoptosis rate of cells was decreased, the expression of hsa_circ_0025853 was up-regulated, and the expression of Drp1 protein and mRNA was down-regulated in H and S+ H groups ( P<0.05). Compared with group H, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of hsa_circ_0025853 was down-regulated, and the expression of Drp1 protein and mRNA was up-regulated in group S+ H ( P<0.05). Conclusion:The mechanism by which hypothermia alleviates the OGD/R injury to SK-N-SH cells is related to the up-regulation of hsa_circ_0025853 expression, thus reducing the expression level of Drp1.

Correlation study of gamma-glutamyl transferase/high density lipoprotein cholesterol ratio,neutrophil/lymphocyte ratio and coronary heart disease / 中国综合临床

Objective:To investigate the correlation between gamma-glutamyl transferase/high-density lipoprotein cholesterol ratio (GHR), neutrophil/lymphocyte ratio (NLR) and coronary heart disease (CHD), and evaluated its pathogenic risk and predictive value for CHD.Methods:A total of 694 patients admitted to our hospital from December 2017 to December 2018 for suspected CHD and coronary angiography were selected. According to the results of coronary angiography,the patients were divided into CHD group ( n=527) and non-CHD group ( n=167). The clinical data of all patients were recorded. Gamma-glutamyl transferase (GGT), high-density lipoprotein cholesterol (HDL-C) and other biochemical indicators were recorded. Neutrophils, lymphocyte count and other hematological indicators were recorded. GHR, NLR and Gensini scores of the patients were calculated. Clinical data and GHR, NLR and other indicators were compared between the two groups. Receiver operating characteristic curve (ROC) was used to evaluate the predictive value of GHR, NLR in CHD, and to determine the optimal cut-off value; Logstic regression analysis was used to investigate the risk factors of CHD.Spearman correlation analysis was used to analyze the correlation between serum OPN, OPG and Gensini score in patients with CHD. Results:The GHR and NLR were 32.59(21.05, 48.24) and 3.53(2.18, 8.46) significantly higher in the CHD group than in the non-CHD group 16.56(10.07, 25.21) and 2.20(1.45, 3.28) respectively, with statistically significant differences ( Z=11.094, 9.055, P<0.05). ROC curve analysis showed that the AUC of NLR and MLR in diagnosing CHD was 0.785 and 0.732( P<0.05). When the critical values of GHR and NLR respectively were 19.805 and 2.678, respectively, the diagnostic efficiency of CHD was the highest, and the sensitivity and specificity were 79.30%, 62.90% and 63.80%, 68.30%, and the AUC of GGT in diagnosing CHD was 0.628. When the critical value was 19.500, the sensitivity and specificity were 80.50% and 39.50%, respectively,the AUC of GHR was greater than that of GGT ( Z=12.973, P<0.05). Multivariate Logistic regression analysis showed that Smoking ( OR=2.887, 95% CI:1.850-4.505, P<0.05), hypertension ( OR=2.009, 95% CI: 1.311-3.080, P<0.05), fasting plasma glucose ( OR=1.109, 95% CI:1.034-1.189, P<0.05), age ≥60 years ( OR=1.567, 95% CI:1.179-2.415, P<0.05), NLR ≥2.687 ( OR=3.152, 95% CI:2.066-4.808, P<0.05) and GHR ≥19.805 ( OR=4.768, 95% CI:3.131-7.262, P<0.05) was an independent risk factor for CHD. After gradually adjustment for risk factors such as smoking, hypertensive, fasting plasma glucose, age ≥60 years and NLR ≥2.687, GHR ≥19.805 was still an independent risk factor for coronary heart disease(OR and 95% CI were 4.620 (3.049-7.000), 4.768 (3.131-7.262), 6.567 (4.408-9.810), 4.768 (3.131-7.262), 4.768 (3.131-7.262), respectively; all P<0.001). Spearman correlation analysis showed that GHR and NLR were positively correlated with Gensini score ( r=0.312, 0.394; all P<0.05). Conclusion:GHR and NLR were positively correlated with the severity of coronary artery disease, which is of significance in the diagnosis of coronary heart disease. NLR ≥2.687 and GHR ≥19.805 were independent risk factors for CHD. GHR was superior to GGT and HDL-C alone in the diagnosis of CHD,and has certain clinical application value

Role of miR-188-5p in oxygen-glucose deprivation and restoration injury to mouse N2a cells: relationship with SENP3 / 中华麻醉学杂志

Objective:To evaluate the role of miR-188-5p in oxygen-glucose deprivation and restoration (OGD/R) injury to mouse neuroblastoma (N2a) cells and its relationship with small ubiquitin-like modifier-specific proteases 3 (SENP3).Methods:N2a cells were cultured and divided into 5 groups ( n=23 each) using a random number table method: control group (group C), OGD/R group, group NC, transfection of mir-188-5p agonist group (group M) and transfection of mir-188-5p inhibitor group group (group I). Cells in group C were cultured routinely.Cells in group NC, group M and group I were transfected with mir-188-5p negative control miRNA, agonist and inhibitor, respectively.N2a cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply to establish the model of OGD/R injury.At 24 h of oxygen-glucose restoration, the cell viability was recorded by the cell counting kit-8 assay, the amount of lactic dehydrogenase (LDH) released was detected, the expression of miR-188-5p and SENP3 mRNA was detected by quantitative real-time polymerase chain reaction, and SENP3 expression was determined by Western blot.The targeting relationship between miR-188-5p and SENP3 mRNA was detected using dual luciferase reporter assay. Results:Compared with group C, the cell viability was significantly decreased, amount of LDH released was increased, and expression of SENP3 and its mRNA was up-regulated in the other 4 groups, miR-188-5p expression was down-regulated in OGD/R and I groups, and miR-188-5p expression was up-regulated in group M ( P<0.05 or 0.01). Compared with group OGD/R, the cell viability was significantly decreased, amount of LDH released was increased, and expression of SENP3 and its mRNA was up-regulated, and miR-188-5p expression was down-regulated in group I, and the cell viability was increased, amount of LDH released was decreased, expression of SENP3 and its mRNA was down-regulated, and miR-188-5p expression was up-regulated in group M ( P<0.05 or 0.01). The dual luciferase reporter assay showed that miR-188-5p could act directly on SENP3. Conclusion:miR-188-5p is involved in OGD/R injury, which is associated with targeted down-regulation of SENP3 expression in N2a cells.

Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP / 中国实验方剂学杂志

Objective:To establish a rapid method to identify <italic>Levisticum officinale </italic>adulterated in<italic> Angelica sinensis</italic> by polymerase chain reaction -restriction fragment length polymophism（PCR-RFLP）. Method:By comparing sequences restriction sites in ribosomal DNA Internal Transcribed Spacer（ITS） of <italic>A. sinensis</italic> and <italic>L. officinale</italic>，the specific restriction site Fnu4HI of <italic>L. officinale</italic> was selected，and the primers for PCR-RFLP reaction were designed. Different <italic>A. sinensis</italic> and <italic>L. officinale</italic> were amplified by PCR. The conditions affecting the PCR-RFLP reaction，such as annealing temperature，primer concentration，cycle number and enzyme digestion reaction time，were optimized，and the accuracy of the method was investigated. The established PCR-RFLP identification method was used to investigate the applicability of <italic>L. officinale </italic>adulterated<italic> in A. sinensis</italic> with different aduleration ratios and different origins. Result:A PCR-RFLP method for identifying <italic>A. sinensis</italic> mixed with <italic>L. officinale</italic> was established. When the annealing temperature was 62 ℃ and the number of cycles was 30，when the <italic>L. officinale </italic>adulterated in<italic> A. sinensis</italic> could be digested by Fnu4H I restriction endonuclease after amplification with specific primers，and the two single DNA bands were detected between 100-500 bp，the <italic>A. sinensis</italic> were all negative. The minimum detection limit of this method for adulterated <italic>L. officinale</italic> in <italic>A. sinensis</italic> was 3%，which could be used for the detection of adulterated <italic>L. officinale</italic> in <italic>A. sinensis</italic>. Conclusion:The established PCR-RFLP identification method is sensitive and accurate in detecting whether there is <italic>L. officinale</italic> in <italic>A. sinensis</italic>，and it provides inspection reference and basis for the quality control of <italic>A. sinensis</italic>，with great significance to ensure the safety of its clinical medication.

Identification of Armeniacae Semen Amarum Adulterated in Persicae Semen by Allele-specific Polymerase Chain Reaction / 中国实验方剂学杂志

Objective:Due to the limitation of traditional identification methods of Chinese medicinal materials， the study established a rapid method to identify Persicae Semen mixed with Armeniacae Semen Amarum by allele-specific polymerase chain reaction （PCR）. Method:By comparing the ribosomal DNA internal transcribed spacer （ITS） gene sequences of Persicae Semen and Armeniacae Semen Amarum， single nucleotide polymorphism （SNP） sites were searched and specific primers were designed. Different Persicae Semen and Armeniacae Semen Amarum samples were amplified by PCR， the effects of annealing temperature， primer concentration and cycle number on the PCR reaction system were optimized， and the specificity and detection limit of this method were investigated. In addition， the established PCR method was used to detect the samples of Persicae Semen mixed with different proportion of Armeniacae Semen Amarum from different sources and producing areas. Result:A specific PCR method for identifying Persicae Semen mixed with Armeniacae Semen Amarum was established. When the annealing temperature was 63 ℃ and the number of primer cycles was 30， only Armeniacae Semen Amarum could be amplified with 432 bp specific band， while Persicae Semen samples did not have this band. The minimum detection limit of this method for Armeniacae Semen Amarum was 0.2 ng， and the detection limit for Armeniacae Semen Amarum adulterated in Persicae Semen was 1%. Conclusion:The established allele-specific PCR method can accurately detect whether there is Armeniacae Semen Amarum in Persicae Semen， which can provide experimental basis for the quality control of Persicae Semen and guarantee the safety of its clinical use.

Metrological Analysis of the Projects in the Field of Forensic Science Funded by the National Natural Science Foundation of China between 2000 and 2019 / 法医学杂志

Objective To understand the trend of funding in the field of forensic science by analyzing the projects funded by the National Natural Science Foundation of China （NSFC）. Methods Based on the data of funded projects in the field of forensic science published on NSFC website in the twenty years from 2000 to 2019, the trend of project number and fund, project type, discipline type, funded institution and personnel were analyzed with metrological methods. The funding theme was analyzed through multi-methods. Results In the twenty years, NSFC funded 352 projects in the field of forensic science, including 9 project types, 47 institutions and 213 persons. General programs （184 projects）, forensic toxicology, pathology, and toxicology analysis （H2301, 188 projects）, and the Academy of Forensic Science （37 projects） had the most funded projects. NSFC funded 174.69 million yuan in the field of forensic science. The Excellent Young Scientists Fund （3.50 million yuan/project）, forensic toxicology, pathology, and toxicology analysis （H2301, 78.64 million yuan）, and Xi'an Jiaotong University （23.78 million yuan） had the most funding. The differences in the number and funding of subsidized projects between the first ten years and the last ten years had statistical significance （P<0.05）. The main targets of funding were toxicology research, evidence-related research, estimation-related research, brain injury research, et al. The field of research mainly funded from 2010 to 2019 was interdisciplinary research. Conclusion Since 2010, the number of project, funding of project, the number of institutions and persons funded by NSFC in the field of forensic science showed an increasing trend. The types of disciplines, types of projects and themes of projects are more diverse, which will promote the long-term development of basic forensic research in China.

Research Progress on Forensic Palynology and Its Application in Forensic Science / 法医学杂志

Forensic palynology is to apply palynology to the field of forensic science, using pollen and spores to solve issues in juridical practice, such as civil and criminal issues. Sporopollens have a small size, wide distribution, diverse morphology, can be easily transferred, have durability, and is not easily noticed. It can provide strong investigation and related evidence for case detection as good trace evidence. The application of palynology in forensic science has achieved certain success, but it is underutilized in most countries. This paper analyzes the evidence value provided by sporopollen, collection of the sporopollen samples, the progress in detection technology and challenges ahead, based on the biological characteristics of sporopollen, combined with recent successful cases in forensic science, to comprehensively discuss the research progress in forensic palynology and its application prospects in forensic science.

Changes and significance of soluble low-density lipoprotein receptor-related protein-1 and regulated upon activation normal T cell expressed and secreted in acute coronary syndrome / 中国综合临床

Objective:To investigate the concentration of soluble low-density lipoprotein receptor-related protein-1(sLRP-1) and regulated upon activation normal T cell expressed and secreted(RANTES) in the plasma of patients with acute coronary syndrome(ACS) and the correlation with the severity of coronary artery disease, and to explore its clinical significance.Methods:From January 2019 to July 2019, 250 patients in the Department of Cardiology of the Affiliated Hospital of Chengde Medical College were selected for prospective study, including 158 patients with ACS, 42 patients with stable angina pectoris (SAP) and 50 patients in the control group.ACS patients included 92 patients with acute myocardial infarction(AMI) and 66 patients with unstable angina pectoris(UAP). The degree of coronary artery lesions was evaluated according to the Gensini score.The patients in the control group were excluded from coronary atherosclerosis by coronary angiography at the same time.The concentrations of sLRP-1 and RANTES in plasma were measured by enzyme linked immunosorbent assay(ELISA).Results:The plasma levels of lg sLRP-1 and RANTES in AMI group ((0.69±0.20) mg/L and(125.17±34.87 )ng/L) and UAP group ((0.62±0.19) mg/L and(118.51±21.46 ) ng/L) were higher than SAP group ((0.42±0.16) mg/L and(99.56±21.46)ng/L) and control group ((0.27±0.08) mg/L and (98.39±19.37)ng/L)(all P<0.05), while lg sLRP-1 and RNATES levels were not significantly different between AMI group and UAP group, SAP group and control group(all P>0.05). The levels of lg sLRP-1 and RANTES in patients with ACS decreased after PCI treatment(all P<0.05). Plasma lg sLRP-1 and RANTES levels were not correlated with Gensini scores in ACS patients(all P>0.05). Conclusion:SLRP-1 and RANTES are involved in the progression of ACS, and there is a certain correlation between them.Myocardial perfusion therapy improves the inflammatory response of coronary artery.