ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion （MCAO） mice by regulating the expression of neutrophil-related chemokines in exosomes， thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free （SPF） C57BL/6J mice were randomly divided into four groups： Sham-operated group， MCAO model group， Huanglian Jiedutang group （6 g·kg^-1）， and Ginaton group （21.6 mg·kg^-1）， with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·（10 g）^-1 for 7 consecutive days， while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days， MCAO surgery was performed. The distal and proximal ends of the right common carotid artery （CCA） were ligated， a small incision was made between the two ligatures， and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling， mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin （HE） staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy， particle size analysis， and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase （MPO） in the brains of mice in each group. ResultsCompared with the sham-operated group， the model group showed decreased neurological function scores （P<0.01）， obvious cerebral infarction （P<0.01）， reduced cerebral blood flow （P<0.01）， neuronal necrosis in the brain， and decreased numbers of Nissl bodies （P<0.01）. The mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were significantly increased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were increased （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were elevated （P<0.01）. Compared with the model group， the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores （P<0.05）， reduced cerebral infarct volume （P<0.01）， restored cerebral blood flow （P<0.01）， reduced necrotic cells in the brain， and increased numbers of Nissl bodies （P<0.01）. In the Huanglian Jiedutang group， the mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were decreased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were reduced （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were decreased （P<0.01）. ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice， thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.

ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion （MCAO） mice by regulating the expression of neutrophil-related chemokines in exosomes， thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free （SPF） C57BL/6J mice were randomly divided into four groups： Sham-operated group， MCAO model group， Huanglian Jiedutang group （6 g·kg^-1）， and Ginaton group （21.6 mg·kg^-1）， with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·（10 g）^-1 for 7 consecutive days， while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days， MCAO surgery was performed. The distal and proximal ends of the right common carotid artery （CCA） were ligated， a small incision was made between the two ligatures， and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling， mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin （HE） staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy， particle size analysis， and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase （MPO） in the brains of mice in each group. ResultsCompared with the sham-operated group， the model group showed decreased neurological function scores （P<0.01）， obvious cerebral infarction （P<0.01）， reduced cerebral blood flow （P<0.01）， neuronal necrosis in the brain， and decreased numbers of Nissl bodies （P<0.01）. The mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were significantly increased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were increased （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were elevated （P<0.01）. Compared with the model group， the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores （P<0.05）， reduced cerebral infarct volume （P<0.01）， restored cerebral blood flow （P<0.01）， reduced necrotic cells in the brain， and increased numbers of Nissl bodies （P<0.01）. In the Huanglian Jiedutang group， the mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were decreased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were reduced （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were decreased （P<0.01）. ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice， thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.

Objective:To observe the effects of electroacupuncture (EA) at acupoints of Shaoyang meridian on mechanical and thermal pain threshold, TRPV1, CGRP mRNA and TRPV1, p-TRPV1, CGRP protein expressions in TG of CM rats; To explore its mechanism.Methods:Totally 24 SD rats were divided into blank group, model group and EA group according to random number table method, with 8 rats in each group. The blank group did not do any treatment; the model group and the EA group were given intraperitoneal injection of nitroglycerin (10 mg/kg) on the 1st, 3rd, 5th, 7th and 9th day, once a day, for 5 times to prepare migraine rat model; after modeling, the model group was given binding and restraint for 30 min, and the EA group was given binding and restraint for 30 min treatment; the changes of basic mechanical and thermal pain thresholds of rats before and after modeling and intervention were observed, the mRNA expression levels of TRPV1 and CGRP in TG rats were detected by qPCR, and the protein expression levels of TRPV1, p-TRPV1 and CGRP in TG rats were detected by Western blot method.Results:After intervention, compared with the model group, the periorbital and plantar mechanical pain thresholds and plantar thermal pain thresholds in EA group increased ( P<0.05), the TRPV1 and CGRP mRNA expressions in trigeminal ganglion decreased ( P<0.01), the protein expressions of TRPV1, p-TRPV1 and CGRP decreased ( P<0.05). Conclusion:EA at acupoints of Shaoyang meridian can obviously increase the mechanical and thermal pain threshold and improve pain sensitization in CM rats. The mechanism would be related to down-regulating the expression levels of TRPV1 and CGRP mRNA and TRPV1, p-TRPV1 and CGRP protein in the TG of CM rats.

ObjectiveTo investigate the possible mechanism of electroacupuncture at Fengchi （GB 20）， Yanglingquan （GB 34） and Waiguan （TE 5） for acute migraine attacks by peroxisome proliferator-activated receptor γ/ nuclear transcription factor-κB pathway in the trigeminal neurovascular system. MethodsForty SD rats were randomly divided into blank group， model group， electroacupuncture group， electroacupuncture plus inhibitor group， and agonist group， 8 rats in each group. Except for the blank group， rats were injected with inflammation decoction intracranial catheter to establish rat models of acute migraine. Thirty minutes after modelling， the electroacupuncture group was given 0.9% NaCl solution by intraperitoneal injection and then electroacupuncture at "Fengchi" （GB 20）， "Yanglingquan" （GB 34）， and "Waiguan" （TE 5） for 30 mins； the electroacupuncture plus inhibitor group was given PPARγ inhibitor T0070907 （1.5 mg/kg） and eclectroacupuncture as the above for 30 mins； the agonist group was given PPARγ inhibitor pioglitazone hydrochloride （10 mg/kg） for 30 mins. Rats in the blank group and the model group were injected intraperitoneally with 0.9% NaCl solution and then bound and restrained for 30 mins. Behavioural scores were evaluated before modelling， 30 mins after modelling （pre-intervention） and post-intervention； after the last behavioural test， PPARγ， NF-κB p65 mRNA content in rat dura mater was detected by real-time fluorescence quantitative PCR； protein expression of PPARγ， NF-κB p65， interleukin 6 （IL-6）， tumour necrosis factor α （TNF-α） in spinal trigeminal nucleus of rats was detected by protein blotting； immunofluorescence double-labelling method was used to detect the fluorescence intensity of PPARγ， NF-κB p65 in spinal trigeminal nucleus of rats. ResultsCompared with the blank group at the same time， the behavioural scores of rats in the remaining groups increased after modelling and after intervention （P＜0.01）. Compared with the model group at the same time， the beha-vioural scores of rats in the electroacupuncture group， electroacupuncture plus inhibitor group， and agonist group decreased after the intervention （P＜0.01）. Compared with the electroacupuncture group at the same time， beha-vioural scores reduced in the agonist group and elevated in the electroacupuncture plus inhibitor group after intervention （P＜0.01）. Compared with the blank group， expression of PPARγ and NF-κB p65 mRNA elevated in the dura mater of rats in the model group， and expression of PPARγ， NF-κB p65， IL-6， and TNF-α protein enhanced in spinal trigeminal nucleus， and the fluorescence intensity of PPARγ and NF-κB p65 enhanced （P＜0.01）. Compared with the model group， PPARγ mRNA expression in dura mater elevated and NF-κB p65 mRNA expression reduced in each intervention group， PPARγ protein expression in spinal trigeminal nucleus enhanced， and NF-κB p65， IL-6， and TNF-α protein expression weakened； in the electroacupuncture group and the agonist group， PPARγ fluorescence intensity enhanced， and the fluorescence intensity of NF-κB p65 weakened in each intervention group （P＜0.05 or P＜0.01）. Compared with the electroacupuncture group， in the electroacupuncture plus inhibitor group， PPARγ mRNA， protein expression and fluorescence intensity reduced， NF-κB p65 mRNA， protein expression and fluorescence intensity elevated， and IL-6 and TNF-α protein expression enhanced； in the agonist group， PPARγ mRNA and protein expression elevated， NF-κB p65 mRNA and protein expression reduced， and IL-6 and TNF-α protein expression decreased （P＜0.05 or P＜0.01）. ConclusionElectroacupuncture at "Fengchi" （GB 20）， "Yanglingquan" （GB 34） and "Waiguan" （TE 5） can can reduce the symptoms of acute migraine attacks in rats， and its mechanism may be related to the up-regulation of PPARγ expression and the reduction of NF-κB expression， thus inhibiting the neurogenic inflammatory response.

Objective To compare the set up errors derived from different registration methods of the X-ray volume imaging (XVI) system for radiotherapy in the treatment of middle/lower-segment esophageal cancer, and to provide a reference for radiation treatment of esophageal cancer. Methods We randomly selected 63 patients with middle/lower-segment esophageal cancer, and obtained their reconstructed XVI images at the first therapy to perform automatic registration with gray-value and bone registration methods. We acquired and compared the three translation errors (along x [left to right], y [head to feet], and z [front to back] axes) and three rotation errors (around the x, y, and z axes) derived from the two registration methods. Results Gray-value registration had significantly smaller translation errors along the x and z axes than bone registration (x azes t = −2.78, z azes t = −2.15, P < 0.05), but there was no significant difference along the y axes (P > 0.05). The rotation errors around the three axes were all smaller than 1°, and were smaller with gray-value registration than with bone registration, but without significant differences (P > 0.05). Conclusion We recommend gray-value registration for radiotherapy in the treatment of middle/lower-segment esophageal cancer. Manual verification or fine-tuning is recommended after automatic registration in clinical practice. Besides translation errors, rotation errors should also be paid attention to.

Objective:To explore the potential mechanism of PD-1 inhibitor P on RIMI from the perspective of immune microenvironment.Methods:To establish a mouse model of radiation-induced myocardial injury (RIMI), twenty C57BL/6 mice were randomly divided into 4 groups, 5 in each group. Group A was the healthy control group; Group B was the PD-1 inhibitor group; Group C was the simple irradiation group, with a heart irradiation of 15 Gy; Group D was the irradiation+ PD-1 inhibitor group. One month after irradiation, the mice were anesthetized and sacrificed. The morphological changes of myocardial tissues were observed by HE staining. The myocardial fibrosis was assessed by Masson staining. CD 3+ , CD 3+ CD 4+ , CD 3+ CD 8 lymphocyte subsets and cytokines (IL-4, IL-6, IL-17A, TNF-α, TGF-β 1 and INF-γ) levels were determined by flow cytometry. The apoptosis rate of myocardial cells was detected by TUNE. Results:One month after irradiation, there was no obvious myocardial fibrosis in group B, and collagen fibers were distributed in the interstitium of myocardial cells in groups C and D. Semi-quantitative analysis results showed that the myocardial collagen volume fraction (CVF) of groups A, B, C and D were (1.97±0.36)%, (2.83±1.03)%, (5.39±0.77)% and (7.72±1.43)%, respectively. The CVF between group A and group B was similar ( P=0.314), and the differences in CVF between the other groups were statistically significant (all P<0.05). Compared with group A, the absolute value and percentage of CD 3+ T lymphocytes were significantly increased in groups B, C and D (all P<0.01). The values in group D were significantly higher than those in group B and group C (all P<0.01); The absolute value and percentage of CD 3+ CD4 T lymphocytes were similar among four groups (all P>0.05); The absolute value and percentage of CD 3+ CD 8 T lymphocytes in group D were significantly higher than those in groups A, B and C (all P<0.001). The expression levels of IL-6, IL-17A, and TGF-β 1 in group D were significantly higher compared with those in groups A, B and C (all P<0.001). The apoptotic index was gradually increased in four groups, and the differences in apoptotic index among four groups were statistically significant (all P<0.001). Conclusion:PD-1 inhibitors can aggravate RIMI by promoting myocardial immune inflammatory response.

Objective The literature study the setup errors of head and neck, thoracic, abdominal and pelvic tumors by megavoltage fan-beam CT based image guidance in TOMO-HD to provide the margin enlarging from clinic target volume (CTV) to planning target volume (PTV) in treatment planning system of TOMO-HD. Methods 103 patients with head and neck (30 patients), thoracic (42 patients), abdominal and pelvic (31 patients) carcinoma were enrolled. Megavoltage fan-beam CT based image guidance in tomotherapy-HD was used to acquire CT scan before every treatment. The left-right (X), superior-inferior (Y), anterior-posterior (Z) and rotation (Fy) setup errors of patients can be obtained from the tomography image automatically restructured by the system. Calculating the systematic error and the random error in the three dimensions and check whether the setup data accord with the normal distribution or not, then acquire the data expand in the three directions. Results According to 2593 fan-beam CT scans, the shift errors (µ ± s) in X, Y, Z and Fy (rotation) of three study group were [(−0.31 ± 2.16) mm、(1.09 ± 3.56) mm、(2.36 ± 2.27) mm, (0.29 ± 0.96)°] (head and neck tumor), [(−0.98 ± 2.95) mm、(0.45 ± 6.86) mm、(3.79 ± 2.47) mm, (0.18 ± 0.60)°] (thoracic cancer) and [(−0.86 ± 2.85) mm、(−1.59 ± 6.91) mm、(5.77 ± 2.40) mm, (0.20 ± 0.68)°](abdominal and pelvic carcinoma). The systematic errors (∑) and random errors (σ) in X, Y, Z dimensions of patients with head and neck, thoracic, abdominal and pelvic tumors were (1.06 mm and 1.84 mm), (1.93 mm and 3.43 mm), (2.41 mm and 2.71 mm), (1.10 mm and 2.56 mm), (3.79 mm and 5.46 mm), (1.38 mm and 1.99 mm) and (1.39 mm and 0.87 mm), (4.98 mm and 5.69 mm), (1.19 mm and 2.05 mm), respectively. Conclusion It is recommended as a reference for image guidance in TOMO-HD according to the frequency distribution of setup errors, for patients with head and neck, chest and abdominal and pelvic tumors, the maximum range of motion in three dimensions are (5.00, 5.00, 5.00) mm, (6.63, 17.25, 16.00) mm and (6.49, 16.24, 13.60) mm.

Objective: To explore the short and mid-term efficacy of device closure of patent foramen ovale (PFO) for treating the patients with PFO combining cryptogenic stroke (CS) and transient ischemic attack (TIA). Methods: A total of 56 PFO patients with CS and TIA receiving device closure in our hospital from 2009-05 to 2015-12 were retrospectively studied. Transthoracic echocardiography (TTE), electrocardiogram (ECG), chest X-ray were examined at 24h, 1 month, 3 and 6 months after theoperation; telephone visit was conducted every 6 months thereafter. Results: There were 54/56 PFO patients combining CS and 2 combining TIA; 53 (94.6%)patients received PFO occluder from Starway medical technology. Aspirin was used for 6 months after the operation. The patients were followed-up for the average of (34.67±23.24) months. No body suffered from post-operative stroke and TIA; no residual shunt was observed. Conclusion: The short and mid-term efficacy of device closure has been satisfactory for treating the patients with PFO combining CS and TIA; its overall clinical value should be further investigated in large population and long-term study.

Objective To study the factor affecting the efficiency of establishment of parthenogenetic stem cells (PESCs)in mice. Methods PESC lines were derived from parthenogenetically activated blastocyst inner cell mass of different mice strains and cultured in different systems.Results The efficiency of establishing the pESC showed no significant difference in hybrid and inbred lines.But the efficiency was increased in culture systems added with ERK inhibitor or knockout serum replacement(KSR).Conclusion The efficiency of establishment of pESC was not directly related with the genetic background of mice but it was closely related to the culture systems.