Analysis of interleukin-36 expression in tears of allogeneic hematopoietic stem cell transplantation patients and its correlation with the ocular surface microenvironment / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To detect the expression of interleukin(IL)-36(α, β, γ)in tears of patients undergoing allogeneic hematopoietic stem cell transplantation(allo-HSCT), investigate its correlation with ocular surface microenvironment, and further analyze the relationship between its expression and ocular graft-versus-host disease(oGVHD).METHODS: Prospective study. A total of 35 patients(70 eyes)underwent allo-HSCT in the hematology department of our hospital in January 2020 were selected, and 35 healthy volunteers(70 eyes)with appropriate age and gender were selected as normal control group. The patients in the allo-HSCT group were followed up 3 times after operation once every 3mo. The subjects with postoperative ocular symptoms were divided into oGVHD and Non-oGVHD group.Ocular surface disease index(OSDI)questionnaire, Schirmer test, tear break-up time(TBUT), corneal fluorescein staining(FL), and conjunctival impression cytology(CIC)was conducted in three groups. Furthermore, the expression levels of IL-36(α,β,γ)in tears were detected by ELISA.RESULTS: In the normal control group, IL-36(α, β, γ)expression levels were 74.32±5.27, 70.02±8.43, 97.41±8.66 pg/mL, respectively; in the allo-HSCT group, IL-36(α, β, γ)baseline expression levels were 77.27±7.03, 74.53±7.53, 100.77±9.74 pg/mL, with no statistically significant differences between the two groups(t=1.648, 1.954, 1.262, all P>0.05). There were no significant differences in IL-36α, IL-36β and IL-36γ in Non-oGVHD group at different time points(P>0.05), while there were significant differences in IL-36α, IL-36β and IL-36γ in oGVHD group at different time points(P<0.05). Compared with Non-oGVHD group, the levels of IL-36α and IL-36β at different time points were significantly increased in oGVHD group(all P<0.05).IL-36(α, β, γ)of oGVHD group was positively correlated with OSDI score, FL and CIC, while it was negatively correlated with TBUT and Schirmer test(all P<0.05).CONCLUSION: Evaluation of levels of tear IL-36(α, β, γ)can be of significance in diagnosing oGVHD after allo-HSCT. IL-36(α, β, γ)is highly expressed in the tears of oGVHD patients before the onset of ocular symptoms, and it is correlated with the ocular surface parameters.

Mechanism of Sleep Deprivation and Cognitive Impairment and Research Progress of Traditional Chinese Medicine Intervention / 中国实验方剂学杂志

Adequate sleep is an important factor to ensure the healthy functioning of the body. A type of chronic sleep diseases characterized by insufficient sleep can be collectively referred to as sleep deprivation （SD）， which is divided into primary and secondary sources in terms of sources. As one of the most frequent types of diseases in recent years， SD has received more and more attention and attention from the whole society. SD can have a wide-ranging and far-reaching impact on cognitive behavior， such as decreased wakefulness， decreased alertness， and inattention， decreased sensory perception， decreased learning and memory capabilities， et al， involving the impact on multiple system functions of the human body， and It is closely related to the occurrence of many diseases， and may cause serious troubles to the normal life of patients and even their relatives and friends. The cognitive impairment caused by SD has been fully verified in clinical tests and various animal behavior experiments， mainly involving pathological damage such as changes in synaptic plasticity， enhanced endoplasmic reticulum stress， circadian rhythm disorders， and energy metabolism imbalance. Western medicine treatments for SD mostly have negative factors such as high side effect and strong addiction. However， Chinese medicine intervention focuses on the overall concept， has long-lasting effectiveness， significant effects， and mild side effects. It has also been widely recognized clinically for improving the complications of sleep disorders. This article reviews the current status and classification of SD research， its pathological mechanisms that lead to cognitive impairment and its molecular-level exploration directions and results. In recent 5 years， the therapeutic effect and experience of traditional Chinese medicine intervention therapy such as compound Chinese medicine， acupuncture and moxibustion as well as auxiliary therapy such as exercise and five sounds， in order to further summarize and clarify the interaction mechanism between SD and cognitive behavior， and provide a theoretical basis for the study of the pathological mechanism of SD disease and future clinical treatment.

Intervention Effect of Modified Tianwang Buxindan on Glucose and Lipid Metabolism in Mice with Chronic Sleep Deprivation Through Orexin A/OX1R / 中国实验方剂学杂志

Objective:to explore the mechanism of modified Tianwang Buxindan in improving abnormal glucose and lipid metabolism in mice with chronic sleep deprivation from the signal pathway of orexin A/ orexin receptor 1（OX1R）. Method:The 50 6-week-old male C57BL/6 mice were randomly divided into blank group ， model group ， estazolam group and Tianwang Buxindan low and high dose groups ，for ten mice of each group. Except the blank group， rats were deprived of sleep for 8 weeks by the method of multi-platform water environment. In the last 4 weeks， Tianwang Buxindan （8.5，17 g·kg-1）and estazolam solution（9.1 mg·kg-1）were given to the stomach， and the blank group and the model group were fed with pure water of the same volume. The food intake and body weight of mice were measured twice a week， on the 49th day， blood samples were collected from the tail vein for glucose tolerance test （GTT），on the 52nd day for insulin tolerance test（ITT）， was used to detect the expression of total cholesterol （TCH）， triglyceride（TG）and free fatty acid（FFA）in serum， and enzyme-linked immunosorbent assay（ELISA）was used to detect the expression of orexin A in serum and hypothalamus. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）and Western blot were used to detect the mRNA and protein expression of OX1R in hypothalamus. Result:After administration， the food intake of mice in each group was different， compared with the blank group， the body weight of model group was significantly reduced（P<0.05）， the glucose tolerance was significantly abnormal， and the TCH， TG， FFA values were significantly increased（P<0.01）. The expression of orexin A in serum and hypothalamus increased significantly（P<0.01）， and the mRNA and protein expression levels of OX1R in hypothalamus increased significantly（P<0.01）. Compared with the model group， the body weight of each group of Tianwang Buxindan was significantly increased（P<0.05）， with better glucose tolerance and insulin sensitivity， TCH， TG， FFA values were significantly reduced（P<0.05，P<0.01）， accompanied by serum and the expression of orexin A in the hypothalamus was significantly decreased（P<0.05，P<0.01）， the mRNA and protein expression levels of OX1R were significantly decreased（P<0.05，P<0.01）. Conclusion:Tianwang Buxindan can protect mice from abnormal glucose and lipid metabolism induced by chronic sleep deprivation， and its mechanism may be related to the down-regulation of orexin A/OX1R signal expression.

Effect and Mechanism of Sulforaphane on G / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of sulforaphane (SFN) on G@*METHODS@#KG1a and KG1cells were treated by different concentrations of SFN for 48 h. Flow cytometry (FCM) was used to analyze the phase distribution of cell cycle. High-throughput sequencing was used to detect the effect of SFN on the expression of cell cycle related genes in KG1a cells. The mRNA expression of P53, P21, CDC2 and CyclinB1 were detected by qPCR. The protein expression of P53, CDC2, P-CDC2 and CyclinB1 were detected by Western blot.@*RESULTS@#Cells in the G@*CONCLUSION@#SFN induces leukemia cells to block in G

Cut-off values of lesion and vessel quantitative flow ratio in de novo coronary lesion post-drug-coated balloon therapy predicting vessel restenosis at mid-term follow-up / 中华医学杂志(英文版)

BACKGROUND@#Drug-coated balloons (DCBs) have emerged as potential alternatives to drug-eluting stents in specific lesion subsets for de novo coronary lesions. Quantitative flow ratio (QFR) is a method based on the three-dimensional quantitative coronary angiography and contrast flow velocity during coronary angiography (CAG), obviating the need for an invasive fractional flow reserve procedural. This study aimed to assess the serial angiographic changes of de novo lesions post-DCB therapy and further explore the cut-off values of lesion and vessel QFR, which predict vessel restenosis (diameter stenosis [DS] ≥50%) at mid-term follow-up.@*METHODS@#The data of patients who underwent DCB therapy between January 2014 and December 2019 from the multicenter hospital were retrospectively collected for QFR analysis. From their QFR performances, which were analyzed by CAG images at follow-up, we divided them into two groups: group A, showing target vessel DS ≥50%, and group B, showing target vessel DS <50%. The median follow-up time was 287 days in group A and 227 days in group B. We compared the clinical characteristics, parameters during DCB therapy, and QFR performances, which were analyzed by CAG images between the two groups, in need to explore the cut-off value of lesion/vessel QFR which can predict vessel restenosis. Student's t test was used for the comparison of normally distributed continuous data, Mann-Whitney U test for the comparison of non-normally distributed continuous data, and receiver operating characteristic (ROC) curves for the evaluation of QFR performance which can predict vessel restenosis (DS ≥50%) at mid-term follow-up using the area under the curve (AUC).@*RESULTS@#A total of 112 patients with 112 target vessels were enrolled in this study. Group A had 41 patients, while group B had 71. Vessel QFR and lesion QFR were lower in group A than in group B post-DCB therapy, and the cut-off values of lesion QFR and vessel QFR in the ROC analysis to predict target vessel DS ≥50% post-DCB therapy were 0.905 (AUC, 0.741 [95% confidence interval, CI: 0.645, 0.837]; sensitivity, 0.817; specificity, 0.561; P < 0.001) and 0.890 (AUC, 0.796 [95% CI: 0.709, 0.882]; sensitivity, 0.746; specificity, 0.780; P < 0.001).@*CONCLUSIONS@#The cut-off values of lesion QFR and vessel QFR can assist in predicting the angiographic changes post-DCB therapy. When lesion/vessel QFR values are <0.905/0.890 post-DCB therapy, a higher risk of vessel restenosis is potentially predicted at follow-up.

Bioinformatics Analysis on Mechanism of Modified Tianwang Buxindan Intervention on Energy Metabolism in Sleep Deprived Rats / 中国实验方剂学杂志

Objective:To detect the changes of functional expression profile of energy metabolism related differential genes in sleep deprived rats before and after intervention by Tianwang Buxindan by microarray sequencing technology， so as to provide possible ideas and theoretical basis for the prevention and treatment of sleep deprivation. Method:The rats were randomLy divided into two groups： the Tianwang Buxindan group and the model group. The Tianwang Buxindan group was given the decoction of Tianwang Buxindan at the dose of 20 g·kg-1， and the model group was given the pure water of equal volume for 14 days. Taking liver， heart and hypothalamus as samples， high-throughput sequencing was used to obtain differential genes. Gene Ontology（Go）classification and kyoto Encyclopedia of Genes and Genomes （KEGG）pathway enrichment analysis were used to construct a co expression network with lncrna. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）was used to detect the mRNA expression levels of neuropeptide Y（NPY）， bispecific phosphatase 1/mitogen-activated protein kinase phosphatase-1（DUSP1/MKP-1）and alpha-L-iduronidase（IDUA）， three key genes with significant differences in energy metabolism. Result:The 321 differentially expressed genes were obtained， 231 of which were up-regulated and 90 down regulated， which mainly promoted the process of lipid metabolism， glucose metabolism and protein metabolism， participated in the synthesis and expression of fibrinogen， vitamin B6 and mesencephalic astrocyte-derived neurotrophic factor（MANF）， and involved mitogen activated protein kinases（MAPK）， p53 gene（p53）， cyclic adenosine monophosphate（cAMP） and other signal pathways. Compared with the model group， the expression of IDUA significantly increased in the Tianwang Buxindan group （P<0.05）， but decreased significantly in NPY and DUSP1（P<0.01）. Conclusion:Tianwang Buxindan can interfere with the energy metabolism mechanism of sleep deprived rats in many ways. By down regulating the mRNA expression level of NPY and DUSP1 genes， it may activate the p38 MAPK signal pathway and affect the lipid metabolism.

Effect of Modified Tianwang Buxindan on Learning and Memory Function and Expression of Inflammatory Factors in Sleep Deprived Rats / 中国实验方剂学杂志

Objective:By studying the effects of Tianwang Buxindan on sleep quality， cognitive function， inflammatory factors and immune-related gene expression in sleep deprivation model rats， explore the effect of Tianwang Buxindan on the learning and memory process under sleep deprivation and its anti-inflammatory effects possible mechanism. Method:The 40 male SPF rats were used to simulate the sleep deprivation model by multi-platform water environment method， and were randomly divided into model group， Tianwang Buxindan group （20 g·kg-1） and estazolam group （0.1 mg·kg-1）， set up a normal group， 10 in each group. A total of 4 weeks of sleep deprivation modeling was performed， and drug intervention was performed 2 weeks later. The model group and the blank group were given equal volumes of pure water. Electroencephalogram （EEG） evaluation of modeling and analysis of sleep structure and quality of rats， Morris water maze positioning navigation and space exploration experiment analysis of learning and memory ability of rats， application of enzyme-linked immunosorbent assay （ELISA） was used to detect the serum inflammatory factor interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， monocyte chemokine-1 （MCP-1） expression， Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA levels of EB virus inducible gene 3 （EBI3）， extracellular signal-regulated kinase 5 （ERK5）， and p21 activated protein kinase 4 （PAK4）. Result:The sleep deprivation model was successfully built. Compared with blank group， the total sleep time， total duration of slow wave sleep and the duration of the first and second phases of slow wave sleep in the model group were significantly shortened （P<0.01）. The incubation period on the upper platform， the total swimming distance and the time to reach the original platform for the first time increased significantly， while the number of times to cross the platform and the target quadrant significantly decreased （P<0.05，P<0.01）. The expression levels of IL-1β，TNF-α and MCP-1 increased significantly （P<0.01）， the mRNA expression levels of EBI3， ERK5 and PAK4 in the hypothalamus of the model group decreased significantly （P<0.01）. Compared with the model group， the sleep quality of the rats in the Tianwang Buxindan group was significantly improved. The total sleep time， the total duration of slow wave sleep and the duration of the first phase of slow wave sleep were significantly increased （P<0.01）. The incubation period on the platform， the total swimming distance and the time to reach the original platform for the first time are shortened， the number of times to cross the platform and the target quadrant time are extended （P<0.05，P<0.01）， IL-1β， TNF-α， MCP-1 expression levels were significantly reduced （P<0.05，P<0.01）， mRNA expression levels of EBI3， ERK5 and PAK4 in rat hypothalamus were significantly increased （P<0.05，P<0.01）. Conclusion:Tianwang Buxindan can improve the sleep quality and learning and memory ability of sleep deprivation model rats， which may be related to the increase of the expression level of related inflammatory factors and its anti-inflammatory effect.

Clinical observation of 15 cases of corneal foreign body injuries caused by chestnut thorns / 国际眼科杂志(Guoji Yanke Zazhi)

Abstract?AIM:To investigate the effective treatment methods of corneal injury caused by chestnut thorns and the factors affecting the disease progression.?METHODS: From Jul.2014 to Oct.2015, the clinical data of 15 patients(15 eyes) with corneal injury caused by chestnut thorns in Ophthalmology Inpatient Department of Wuhan Tongji Hospital was retrospective analyzed. The patients without fungal keratitis were treated with the surgery of removing chestnut thorn from cornea and antifungal drugs. For the patients complicated with fungal keratitis, besides surgery of removing chestnut thorn and antifungal drugs, anterior chamber irrigation and corneal stroma injection with fluconazole solution were given to treat the disease.If necessary, amniotic membrane transplantation or keratoplasty was also given to the patients complicated with fungal keratitis. After that, the effectiveness of those methods and the factors affecting progression were analyzed.?RESULTS:For 11 patients without fungal keratitis, the average time between corneal injury and receiving treatment at Tongji Hospital was 1-7 (2.42±2.15) d and for 4 patients complicated with fungal keratitis, the average time was 3-30 (18.25±4.35)d.Among 15 cases, statistics suggested that the average number of chestnut thorn in patients complicated with fungal keratitis was 4.5, and all the chestnut thorn penetrated the cornea into the anterior chamber.The average number of chestnut thorn in patients without fungal keratitis was 3.5, and the proportion of chestnut thorn penetrated the cornea into the anterior chamber was 28.5%.After treatment, all patients had no new fungal keratitis or other complications.Those results indicated that the different treatments for the patients with or without fungal keratitis were all effective.?CONCLUSION:The factors affecting the progression of cornea foreign body injury caused by chestnut thorn are the number of chestnut thorn, whether chestnut thorn penetrate the cornea into the anterior chamber, time since injury, active anti -fungal therapy. If patients complicated with fungal keratitis could be treated with antifungal agents and anterior chamber irrigation or corneal stroma injection using fluconazole solution without delay, the progress of fungal keratitis could be effectively controlled, and favorable conditions for further therapy such as amniotic membrane transplantation or keratoplasty could be provided.

Effect of carbon disulfide on mitochondrial respiratory chain in spermatogenic cells in male rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the effect of carbon disulfide (CS(2)) on the mitochondrial respiratory chain in testicular spermatogenic cells in male rats and to explore the possible mechanism of reproductive system damage caused by CS(2) in male rats.</p><p><b>METHODS</b>Twenty-four male Sprague-Dawley rats (clean grade) were randomly divided into four groups: three CS(2) exposure groups (CS(2) concentrations: 50, 250, and 1250 mg/m(3)) and a control group. The rats in CS(2) exposure groups were exposed to CS(2) by static inhalation for 10 weeks (2 h/d, 5 d/w), while the rats in control group were exposed to air. Then, all rats were sacrificed by decapitation; testicular tissues were collected, and mitochondrial protein in spermatogenic cells were extracted; the levels of mitochondrial respiratory chain enzyme complex I∼V were measured by enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>Compared with the control group, all CS(2) exposure groups had significantly increased levels of mitochondrial respiratory chain enzyme complex I∼V in spermatogenic cells (P < 0.05). There were no significant differences in the levels of respiratory chain enzyme complex I∼IV between the CS(2) exposure groups (P < 0.05), but the level of respiratory chain enzyme complex V rose significantly as the concentration of CS(2) increased (P<0.05).</p><p><b>CONCLUSION</b>Various levels of CS(2) exposure may increase the levels of mitochondrial respiratory chain enzyme complex in testicular spermatogenic cells among male rats, thus affecting the normal oxidative phosphorylation in mitochondria.</p>

Effect of carbon disulfide on mitochondrial apoptosis-related proteins in cytoplasm of testicular tissue among male rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the role of mitochondrial pathway in the apoptosis of spermatogenic cells induced by inhalation of carbon disulfide in male rats.</p><p><b>METHODS</b>Twenty-four male Sprague-Dawley rats (clean grade) were divided into four groups according to their body weights: three CS(2) exposure groups (CS(2) concentrations: 50, 250, and 1250 mg/m(3)) and a control group. The rats in CS(2) exposure groups were exposed to CS(2) by static inhalation for 10 weeks (2 h/d, 5 d/w), while the rats in control group were exposed to air. Then, all rats were sacrificed by decapitation; testicular tissues were collected, and cytoplasmic proteins were extracted; the levels of apoptosis-inducing factor (AIF), cytochrome c (cyto c), Bcl-2, Bax, procaspase-9, and procaspase-3 were measured by Western blot, and the activities of caspase-9 and caspase-3 were measured using a test kit.</p><p><b>RESULTS</b>Compared with the control group, all CS(2) exposure groups had significantly increased levels of cyto c in the cytoplasm of testicular tissue (P<0.05); in the 250 mg/m(3) CS(2) exposure group, the Bax/Bcl-2 ratio and activities of caspase-9 and caspase-3 increased significantly (P<0.05), and the content of procaspase-9 and procaspase-3 decreased significantly (P<0.05); in the 1250 mg/m(3) CS(2) exposure group, the relative expression levels of Bax and AIF in cytoplasm increased significantly (P<0.05), and the expression level of Bcl-2 decreased significantly (P<0.05).</p><p><b>CONCLUSION</b>Mitochondrial pathway plays an important role in the CS(2)-induced apoptosis of spermatogenic cells in testicular tissue among male rats.</p>

Studies on purification of methamidophos monoclonal antibodies and comparative immunoactivity of purified antibodies / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To purify Methamidophos (Met) monoclonal antibodies with two methods and compare immune activity of purified antibodies.</p><p><b>METHOD</b>Caprylic acid ammonium sulphate precipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method were used to purify Met monoclonal antibodies, UV spectrum scanning was used to determine protein content and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linked immunosorbent assay (ELISA) was used to determine immune activity of purified antibodies.</p><p><b>RESULTS</b>Antibody protein content and recovery rate with CAASP method were 7.62 mg/mL and 8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mL and 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASP method. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was 181.26 microg/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01 microg/mL and 1.03 microg/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82 microg/mL, and the linear working range and LOD were 10.91-11412.29 microg/mL and 3.42 microg/mL, respectively.</p><p><b>CONCLUSION</b>Antibodies purified by SPA method are better than those by CAASP method, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelled testing paper for analyzing Met residue in vegetable and drink water.</p>