ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design（QbD） and to assess its health risk. MethodsTaking whole iron content， Fe²⁺ dissolution content and looseness as critical quality attributes（CQAs）， and calcination temperature， calcination time， spreading thickness and particle size as critical process parameters（CPPs） determined by the failure mode and effect analysis（FMEA）， the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation（AHP-CRITIC） hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry， and the health risk assessment was carried out by daily exposure（EXP）， target hazard quotient（THQ） and lifetime cancer risk（LCR）， and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃， calcination time of 1 h， particle size of 0.2-0.5 cm， spreading thickness of 1 cm， and vinegar quenching for 1 time［Haematitum-vinegar（10：3）］. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1， while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10^-6-1×10^-4， and the LCR of the rest was<1×10^-6， and the LCRs of calcined Haematitum were all<1×10^-6， indicating that the carcinogenic risk of calcined Haematitum was low， but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg were formulated as 1 014， 25， 17， 27， 7 mg·kg^-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible， and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.

ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design（QbD） and to assess its health risk. MethodsTaking whole iron content， Fe²⁺ dissolution content and looseness as critical quality attributes（CQAs）， and calcination temperature， calcination time， spreading thickness and particle size as critical process parameters（CPPs） determined by the failure mode and effect analysis（FMEA）， the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation（AHP-CRITIC） hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry， and the health risk assessment was carried out by daily exposure（EXP）， target hazard quotient（THQ） and lifetime cancer risk（LCR）， and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃， calcination time of 1 h， particle size of 0.2-0.5 cm， spreading thickness of 1 cm， and vinegar quenching for 1 time［Haematitum-vinegar（10：3）］. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1， while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10^-6-1×10^-4， and the LCR of the rest was<1×10^-6， and the LCRs of calcined Haematitum were all<1×10^-6， indicating that the carcinogenic risk of calcined Haematitum was low， but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg were formulated as 1 014， 25， 17， 27， 7 mg·kg^-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible， and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.

Objective:To investigate the correlation between the number of circulating tumor cells (CTC) in peripheral blood and clinicopathological features of patients with breast cancer.Methods:The clinical data of 104 breast cancer patients at Guangzhou Panyu Central Hospital between January 2017 and May 2020 were retrospectively analyzed. The number of CTC in peripheral blood, the levels of serum tumor markers [alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), carbohydrate antigen (CA)125, CA153] were detected. In blood samples, the number of CTC ≥ 2/ml was defined as CTC positive. Immunohistochemistry was used to analyze the protein expression of Ki-67 in tumor tissues. The association of CTC with clinicopathological features, serum tumor markers and Ki-67 protein expression was also analyzed.Results:The CTC positive rate was 80.77% (84/104). There were statistically significant differences in composition of whether there was vascular tumor thrombus (χ 2 = 0.860, P = 0.009), axillary lymph node metastasis (χ 2 = 12.382, P<0.01), N staging ( P = 0.002) and TNM staging (χ 2 = 7.698, P = 0.006) between patients with CTC positive and negative. However, there were no statistically significant differences in composition of age ( t = 0.634, P = 0.528), tumor quadrant (χ 2 = 6.523, P = 0.163), molecular subtyping (χ 2 = 4.164, P = 0.384), histological grade (χ 2 = 1.901, P = 0.387), T staging ( P = 0.099) and whether there was nerve invasion (χ 2 = 0.092, P = 0.761). The levels of serum CEA and CA125 in CTC positive patients were higher than those in CTC negative patients [median ( P25, P75): 2.50 ng/ml (2.21 ng/ml, 2.92 ng/ml) vs. 1.89 ng/ml (1.61 ng/ml, 2.35 ng/ml); 13.81 U/ml (11.79 U/ml, 16.28 U/ml) vs. 11.17 U/ml (8.91 U/ml, 12.80 U/ml); all P < 0.05], and CTC was positively correlated with serum CEA and CA153 levels ( r = 0.520, P<0.01; r = 0.497, P<0.01); CTC was not related to Ki-67 protein expression (χ 2 = 0.512, P = 0.474). Conclusion:The number of CTC in peripheral blood is closely related to clinical staging, lymph node or hematogenous metastasis, tumor markers CEA and CA153 levels of breast cancer. The increased number of CTC may cause tumor progression and metastasis.

Objective:To explore the consistency of peripheral whole blood and venous serum procalcitonin (PCT) levels, and the value of peripheral whole blood PCT in evaluating pediatric bacterial infection.Methods:This multicenter cross-sectional parallel control study was conducted in 11 children′s hospital. All the 1 898 patients older than 28 days admitted to these hospitals from March 2018 to February 2019 had their peripheral whole blood and venous serum PCT detected simultaneously with unified equipment, reagent and method. According to the venous serum PCT level, the patients were stratified to subgroups. Analysis of variance and chi-square test were used to compare the demographic characteristics among groups. And the correlation between the peripheral blood and venous serum PCT level was investigated by quantitative Pearson correlation analysis.The PCT resultes were also converted into ranked data to further test the consistency between the two sampling methods by Spearman′s rank correlation test. Furthermore, the ranked data were converted into binary data to evaluate the consistency and investigate the best cut-off of peripheral blood PCT level in predicting bacterial infection.Results:A total of 1 898 valid samples were included (1 098 males, 800 females),age 27.4(12.2,56.7) months. There was a good correlation between PCT values of peripheral whole blood and venous serum ( r=0.97 , P<0.01). The linear regression equation was PCT?venous serum=0.135+0.929×PCT peripheral whole blood. However, when stratified to 5 levels, PCT results showed diverse and unsatisfied consistency between the two sampling methods ( r=0.51-0.92, all P<0.01). But after PCT was converted to ordinal categorical variables, the stratified analysis showed that the coincidence rate of the measured values by the two sampling methods in each boundary area was 84.9%-97.1%. The dichotomous variables also showed a good consistency (coincidence rate 96.8%-99.3%, Youden index 0.82-0.89). According to the severity of disease, the serum PCT value was classified into 4 intervals（<0.5、0.5-<2.0、2.0-<10.0、≥10.0 μg/L）, and the peripheral blood PCT value also showed a good predictive value (AUC value was 0.991 2-0.997 9). The optimal cut points of peripheral whole blood PCT value 0.5、1.0、2.0、10.0 μg/L corresponding to venous serum PCT values were 0.395, 0.595, 1.175 and 3.545 μg/L, respectively. Conclusions:There is a good correlation between peripheral whole blood PCT value and the venous serum PCT value, which means that the peripheral whole blood PCT could facilitate the identification of infection and clinical severity. Besides, the sampling of peripheral whole blood is simple and easy to repeat.

Objective To investigate the efficacy and clinical value of balloon-assisted clipping for the treatment of giant unruptured intracranial aneurysms of internal carotid artery. Methods Patients with giant unruptured intracranial aneurysm of intracranial segment of internal carotid artery treated with balloon-assisted clipping in the Department of Neurosurgery, Xiangya Hospital, Central South University from September 2017 to May 2018 were enrolled retrospectively. The proximal internal carotid artery or the aneurysm neck were temporarily blocked by balloon, and then the aneurysm was clipped in the hybrid operating room. Demographic data, preoperative symptoms, aneurysm characteristics, position of balloon placement, intraoperative angiography, complications, and follow-up results were collected. Results A total of 12 patients with giant (diameter >2 cm) unruptured intracranial aneurysm of intracranial segment of internal carotid artery were enrolled. They were all successfully clipped using balloon-assisted clipping in the hybrid operating room. Among them, 1 was located in the ophthalmic segment, 3 in the supraclinoid segment, 4 in the posterior communicating segment, 2 in the anterior choroidal artery segment, and 2 in the bifurcation of the internal carotid artery. The balloons were placed in the proximal end of internal carotid artery in 9 cases and in the neck of aneurysm in 3 cases. Intraoperative angiography showed that 12 aneurysms were completely occluded; 1 had severe stenosis of parent artery, and 1 had mild stenosis. Postoperative complications included cerebral infarction in 1 case, temporary diabetes insipidus in 1 case (returned to normal 1 week after operation), hemiplegia in 1 case, and epilepsy in 1 case. Glasgow Outcome Scale score at discharge showed 5 in 9 cases, 4 in 2 cases, and 3 in 1 case. The patients were followed up for 2.3 to 12 months after operation (median 7.5 months). Reexamination of CT angiography showed no recurrence of aneurysm. Glasgow Outcome Scale score was 5 in 11 cases and 4 in 1 case. Conclusions The use of balloon-assisted clipping technique in the hybrid operating room for the treatment of giant intracranial segmental aneurysms of the internal carotid artery is safe and effective, and has a good long-term outcome.

Objective To investigate the clinical effects and influencing factors of the outcomes of early microsurgical treatment in patients with intracerebral hematoma from ruptured intracranial aneurysm. Methods From 2010 to 2016, patients with intracerebral hematoma from ruptured intracranial aneurysm admitted to the Department of Neurosurgery, Xinyu People's Hospital were enrolled retrospectively. The demographic data, Hunt-Hess grade,Glasgow coma scale(GCS)score,imaging data,and procedure-related complications were collected. Glasgow outcome scale (GOS) score was used to evaluate the outcomes. Four to 5 were defined as good outcome and 1 to 3 were defined as poor outcome. The Hunt-Hess gradesⅡ-Ⅲ were used as the low-grade group and the Ⅳ-Ⅴ grades were used as the high-grade group. The survival rate and quality of life of both groups of patients were compared according to the GOS scores. Results A total of 36 patients were enrolled during the study, including 32 with subarachnoid hemorrhage and intracerebral hematoma and 4 with simple intracerebral hematoma. Hunt-Hess grade was grade Ⅱ in 2 cases, Ⅲ in 18 cases, Ⅳ in 14 cases, and Ⅴ in 2 cases. Distribution of responsible aneurysms:18 patients in middle cerebral artery, 9 in anterior communicating artery, 6 in anterior cerebral artery, 3 in posterior communicating artery, including 4 patients with multiple aneurysms. All patients underwent aneurysm clipping+hematoma removal under the general anesthesia within 36 h after onset,24 of them were treated with decompressive craniectomy. One patient died of severe brain swelling after intraoperative reruptureof the aneurysm,1 died of postoperative massive cerebral infarction, and 1 died of severe pulmonary infection and diabetes after giving up further treatment. Thirty-three survivors were followed up for 1 year, 29 had good outcome(80.5％) and 7 had poor outcome (19.5％). There were significant differences in survival rate and quality of life between the low-grade group and the high-grade group (P=0.001). There were significant differences in the Hunt-Hess grade, baseline GCS score, and proportion of patients receiving decompressive craniectomy between the good outcome group and the poor outcome group.Conclusion The Hunt-Hess grade, baseline GCS score, and decompressive craniectomy were the influencing factors of the outcomes in patients with intracerebral hematoma from ruptured intracranial aneurysm. Removal of hematoma and aneurysm clipping should be performed as early as possible,and decompressive craniectomy should be performed if necessary.

Objective To investigate the associations of serum soluble CD40 ligand (sCD40L) levels with stroke risk,severity,and infarct volume.Methods Consecutive inpatients with acute ischemic stroke were recruited as a patient group.Healthy subjects were used as a control group.The demographics,vascular risk factors,and clinical data were collected from the patient group and control group.The serum sCD40L levels were measured by enzyme linked immunosorbent assay.According to the baseline National Institutes of Health Stroke Scale (NIHSS) scores,they were divided into a mild stroke group (＜ 8) and a moderate to severe stroke group (≥ 8).According to the median of infarct volume,the patients with ischemic stroke were divided into either a large infarction group or a small infarction group.Results A total 106 patients with acute ischemic stroke were recruited,including 47 females (44.3％) and 59 males (55.7％),and the mean age was 71.31 ± 11.27 years.There were 86 healthy subjects in the control group,including 41 females (47.7％) and 45 males (52.3％),the mean age was 73.56±9.32 years;there were.41 patients (38.7％) in large infarction group (≥1.8 cm3) and 65 (61.3％) in the small infarction group (＜1.8 cm3);there were 69 patients (65.1％) with mild stroke and 37 (34.9％) with moderate to severe stroke.The baseline serum sCD40L level in the patient group was significantly higher than that in the control group (5.61 ± 1.68 mg/L vs.3.56 ± 1.32 mg/L;t =9.236,P ＜0.01),the serum sCD40L level at day 14 after admission (4.19 ± 1.45 mg/L) in the patient group was significantly lower than the baseline level (P ＜0.01),but it was still higher than the control group (P ＜ 0.01).Multivariate logistic regression analysis showed that the higher low-density lipoprotein cholesterol (odds ratio [OR] 3.358,95％ confidence interval [CI] 2.681-4.056;P＜0.001) and serum sCD40L (OR 5.103,95％ CI 2.317-8.903;P＜0.001) levels were the independent risk factors for ischemic stroke;the higher serum sCD40L level (fourth vs.first quartile,OR 4.017,95％ CI 1.608-10.037;P =0.003),large atherosclerotic stroke (OR 2.321,95％ CI 1.014-5.314;P =0.046),cortical-subcortical infarcts (OR 2.679,95％ CI 1.111-6.460;P =0.028),and larger infarct volume (OR 3.216,95％ CI 1.398-7.395;P=0.006) were the independent risk factors for moderate to severe stroke;the higher serum sCD40L level (fourth vs.first quartile,OR 3.142,95％ CI 1.274-7.745;P =0.013),large atherosclerotic stroke (OR 2.956,95％ CI 1.299-6.767;P =0.010),cortical-subcortieal infarcts (OR 4.750,95％ CI 1.909-11.818;P ＜0.001),and baseline NIHSS score ≥8 (OR 8.509,95％ CI 3.432-21.094;P ＜ 0.001) were the independent risk factors for large infarction.Conclusion The serum sCD40L levels are closely associated with the risk,severity and infarct volume of ischemic stroke.

Nerve agent not only inhibit acetylcholinesterase ( AChE) at an early stage, but also induce prolonged and progressive neuroinflammation and delayed neurodegeneration.Recently, the US National Institute of Health ( NIH) has sponsored some major programs of toxic mechanisms and treatment of nerve agents, which aims at the development of quick and effective treatment to acute intoxication and delayed effect.The experimentally effective new antidotes mainly include AChE-targeting drugs, broad-spectrum reactivators and scavengers, antiinflamatory and nerve protection drugs.

Objective To compare the changes in energy metabolism in 2-chloroethyl ethryl sulfide(CEES)-poisoned bronchial epithelial cell 16HBE cultured in media at different glucose concentrations .Methods Bronchial epithelial cell 16HBE was cultured in high (4.5 mg/ml) or low (1.1 mg/ml) glucose medium and exposed to a sulfur mustard simulant CEES of 0.2, 0.5, 1.0 mmol/L.Cell growth and cytotoxicity were tested using MTS .ATP, ADP and AMP were detected by HPLC and the value of ATP/ADP, total adenine nucleotides ( TAN) and energy charge ( EC) was subsequently calculat-ed.Mitochondrial oxidative phosphorylation-related proteins, COX-10 and ISCU, were detected using Western blotting . Rhodamine 123 was applied to detect the mitochondrial membrane potential using flow cytometry .Results Low glucose accelerated the growth and energy metabolism of 16HBE cells in regular culture , and the contens of ADP , TAN, COX-10 and ISCU in low glucose group were significantly higher than those in high glucose group .CEES exposure (≥0.5 mmol/L) significantly affected cell viability in both high and low glucose groups , with significant difference between the two groups exposed to 1.0 mmol/L CEES.In high glucose group, 24 h after 0.5 or 1.0 mmol/L CEES exposure, the contents of ATP, ADP and TAN were significantly increased , while ATP/ADP and EC decreased .In low glucose group , ADP, AMP and TAN significantly decreased, while ATP/ADP and EC increased 24 h after 1.0 mmol/L CEES exposure.The mi-tochondrial membrane potential (MMP) also changed differently after 0.5 mmol/L CEES exposure.MMP in high glucose group marginally increased at 3 h, and significantly increased at 8-12 h (P<0.05), and returned to normal at 24 h. MMP in low glucose group showed a transient decrease at 5 h (P<0.01), and back to normal at 8 h.The protein levels of COX-10 and ISCU were significantly increased in high glucose group 24 h after 0.5-1.0 mmol/L CEES exposure , but sig-nificantly decreased in low one 24 h after 1.0 mmol/L CEES exposure .Conclusion When 16HBE is cultured at a high or low glucose concentration , the cell growth, stress responses and energy metabolism including MMP , COX-10, ISCU and ATP production are in different status before or after CEES exposure .High glucose could protect against CEES exposure .

Objective To explore the effect of 2-chloroethyl ethyl sulfide(CEES) poisoning on keratinocyte migration and the regulatory role of microRNA(miR)-34a.Methods MTS was used to detect the viability of cells exposed to CEES in order to select an appropriate dose of CEES exposure in this in vitro model.The protein level of keratin 5 and keratin 10 was detected to assess cell differentiation status .Scratch assay was applied to evaluate cell migration ,and miR-34a silencing in keratinocytes was achieved by transfecting chemically synthesized miR-34a specific miRNA inhibitor.t-ERK1/2 and p-ERK1/2 levels closely related to cell migration were detected using Western blotting .Results An in vitro CEES exposure model of keratinocytes was established at the optimal concentration of 0.5 mmol/L CEES in the viability test , and this dose was chosen to evaluate cell migration changes .The migration of cells was significantly inhibited 24 h after CEES exposure , accompanied by no changes in morphology and keratin 5/10 levels.Silencing of miR-34a significantly increased the migration of cells exposed to CEES , which could be blocked by adding 5 μmol/L U0126 , an ERK1/2 phosphorylation selective inhibitor.Conclusion Silencing of miR-34a can significantly increase keratinocyte migration and partially reverse the inhibition of CEES-caused migration , which could be mediated by ERK 1/2 pathway activation .