OBJECTIVEThis study investigated the effect of hyperbaric oxygenation (HBO) on neural stem cells (NSCs) and myelin in neonatal rats following hypoxic-ischemic brain damage (HIBD) and aimed to explore the possible mechanism of the protective effect of HBO on HIBD.

METHODSSeven-day-old Sprague-Dawley rat pups were randomly assigned into 4 groups: Normal control, HIBD, hyperbaric air (HBA), and HBO groups (n=30 each). The HIBD model was produced by permanent occlusion of the left common carotid artery and 2 hrs hypoxemia exposure (8% O2 at 37 degrees C). HBA and HBO treatment was administered (2 ATA, once daily for 7 days) in the HBA and HBO groups respectively 1 hr after HIBD. BrdU immunohistochemistry was used to detect the NSCs in the sub-ventricle zone (SVZ) of the lateral ventricle and the dentate gyrus (DG) of the hippocampus. The myelin damage was assessed by myelin basic protein (MBP) immunostaining.

RESULTSThe BrdU-positive cells in the SVZ and the DG of the ischemic hemisphere in the HIBD group were dramatically decreased compared with those of the Normal control group at 3 weeks post-HIBD (P < 0.01). The HBO treatment resulted in an increase of BrdU-positive cells in the DG from 153.7 +/- 37.0 to 193.7 +/- 38.8 (P < 0.05). The nestin expression in the HIBD and HBA groups was reduced compared with that in the Normal control group. There was no difference in the nestin expression between the HBO and the Normal control groups. Hypoxia-ischemia (HI) led to marked myelin damage at 1 week post-HIBD. HBO or HBA treatment alleviated the damage.

CONCLUSIONSThe HBO treatment can result in the proliferation of BrdU-positive cells and alleviate the myelin damage following HIBD in neonatal rats, thereby offering neuroprotectivity against HI insults.

Animals ; Animals, Newborn ; Bromodeoxyuridine ; metabolism ; Female ; Hyperbaric Oxygenation ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; therapy ; Immunohistochemistry ; Intermediate Filament Proteins ; analysis ; Male ; Mice ; Myelin Basic Protein ; analysis ; Nerve Tissue Proteins ; analysis ; Nestin ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology

OBJECTIVETo compare biological characteristics between nucleus pulposus and annulus fibrosus cells in vitro model.

METHODSFive New Zealand white rabbits (2 to 3 kg, either gender) were isolated nucleus pulposus and annulus fibrosus under sterilized condition, then cultured in nutrient solution with 15% FBS and DMEM/F12 (1:1) by enzyme digestion combined with tissue block method. When 90% cells fused, subcultring were performed. Cell morphology were observed by inverted phase contrast microscope, cell viability were detected by trypan blue staining, histological were observed by a toluidine blue and HE staining, cell proliferation were tested by MTT method, then the cell morphology, viability, proliferation between nucleus pulposus and annulus fibrosus were compared.

RESULTSThere were no obviously differences between nucleus pulposus and annulus fibrosus in original and the first strain. Physalides were appeared in annulus fibrosus on the second generation. The strapping time was later, and activity was lower in nucleus pulposus than annulus fibrosus. The growth of cell proliferation in nucleus pulposus was lower than annulus fibrosus from the ninth day.

CONCLUSIONThe cell activity in annulus fibrosus is higher than nucleus pulposus. Digenerative disc disease may caused by recession of nucleus pulposus,local biomechnical changes, furether caused structure change and function loss of annulus fibrosus.

Animals ; Cell Proliferation ; Cell Survival ; Disease Models, Animal ; Female ; Humans ; Intervertebral Disc ; cytology ; Intervertebral Disc Degeneration ; physiopathology ; Male ; Rabbits

OBJECTIVESTo understand the value of measuring neonatal cerebral regional oxygen saturation (rSO2) using near infrared spectroscopy (NIRS) in assessing cerebral oxygenation, to establish the normal range of neonatal cerebral rSO2 and to collect data of the changes of cerebral rSO2 under certain disease status.

METHODSNine large hospitals participated in the multicenter randomized clinical trial from Jan 2007 to Apr 2008. Using the NIRS human tissue oximeter (TSAH-100) independently developed in China, the cerebral rSO2 of 223 normal full-term and 95 otherwise healthy preterm neonates without any special disease, was detected at 1, 2 and 3 days after birth, respectively. The cerebral rSO2 of 102 neonates with diseases which may affect the cerebral oxygenation, was also detected during the severe phases. The pulse oxygen saturation (SpO2) measured at the finger tip, and also the arterial oxygen saturation (SaO2) measured by blood gas analysis, which could indicate the oxygen supply of the whole body, were obtained simultaneously. The correlations among cerebral rSO2, pulse SpO2 and arterial SaO2 were analyzed.

RESULTS(1) The cerebral rSO2 of the normal full-term neonates was (62+/-2)%. Cerebral hypoxia was defined as rSO2 lower than 58%. The cerebral rSO2 of the normal full-terms was steady at 1, 2 and 3 days after birth respectively, without any significant differences among them (F=0.610, P>0.05). The cerebral rSO2 of the neonates with diseases was (55+/-7)%, which was significantly lower than that of the normal full-term neonates (t=15.492, P<0.05). (2) The cerebral rSO2 was positively correlated with the SpO2 (r=0.74, P<0.01) and the SaO2 (r=0.71, P<0.01). (3) Under some special diseases, the changes of cerebral rSO2 was asynchronous with those of the SpO2: (1) For 18 cases under severe cerebral damages or under relatively low hemoglobin concentration, the cerebral rSO2 was significantly low (50%-58%), but the SpO2 was still normal (above 90%). (2) During the recovery of some critically ill neonates, the increase of cerebral rSO2 was lagged as compared with that of pulse SpO2. Especially, during the severe phases of 6 cases with multi-organ failure, the SpO2 and the cerebral rSO2 were both significantly low (55%-80% for SpO2, and 44%-50% for cerebral rSO2); when the diseases were alleviated, although the SpO2 recovered to above 85%, the cerebral rSO2 was still significantly low (around 50%). (3) In 3 cases, during the severe phases of serious hypoxic-ischemic encephalopathy (HIE), the cerebral rSO2 significantly increased to 70%-72%, which was significantly higher than the normal value (62%).

CONCLUSIONSThe range of cerebral rSO2 of the normal full-term neonates was (62+/-2)%. Cerebral oxygenation can be externally indicated by the rSO2 noninvasively and continuously measured by NIRS, which was positively correlated with traditional pulse SpO2 and arterial SaO2. In some special diseases, the rSO2 measured by NIRS can be helpful for clinical diagnoses and treatments.

Birth Weight ; Brain ; metabolism ; Female ; Humans ; Hypoxia, Brain ; diagnosis ; Infant, Newborn ; Male ; Oximetry ; methods ; Oxygen ; analysis ; Spectroscopy, Near-Infrared

In addition to providing energy for cells, mitochondria also participate in calcium homeostasis, cell information transfer, cell apoptosis, cell growth and differentiation. Therefore, maintaining mitochondrial homeostasis is very crucial for the body to carry out normal life activities. Ubiquitination, a post-translational modification of proteins, is involved in various physiological and pathological processes of cells by regulating mitochondrial homeostasis. However, the mechanism by which ubiquitination regulates mitochondrial homeostasis has not been summarized, especially the effect of Parkin protein on cardiovascular diseases. In this paper, the specific mechanism of mitochondrial homeostasis regulated by ubiquitination of Parkin protein is discussed, and the influence of mitochondrial homeostasis imbalance on cardiovascular diseases is reviewed, with a view to providing potential therapeutic strategies for the clinical treatment of cardiovascular diseases.

This paper reports the clinical data of a patient with recurrent metastatic parathyroid carcinoma. The causes, clinical manifestation, diagnose, treatment and prognosis of parathyroid carcinoma were discussed in order to perfect the experience of diagnosis and treatment and improve the survival rate of such patients.

Objective:To construct a simple, precise and personalized comprehensive nomogram for prediction the risk of multidrug-resistant tuberculosis (MDR-TB) and to evaluate its prediction value among individuals with previous tuberculosis history (PTBH).Methods:A matched case-control study (1∶2 ratios) was performed in 1 881 patients with PTBH treated in 12 designated tuberculosis hospitals in Hangzhou City between January 1, 2005 and December 31, 2019, and there were 1 719 patients in training set, and 162 in validation set. A multivariable Cox regression analysis was used to evaluate independent predictors for the incident of MDR-TB in individuals with PTBH. A comprehensive nomogram was developed based on the multivariable Cox model. The accuracy of the prediction was assessed using concordance index (C-index), calibration curve and area under the receiver operator characteristic (ROC) curve.Results:The nomogram constructed based on the multivariable Cox regression model incorporated 10 independent predictors of the risk of MDR-TB. A history of direct contact (grade 1, 0-100.0 points) ranked on the top of all risk factors, followed by duration of positive sputum culture (grade 2, 0-84.5 points), unfavorable treatment outcome (grade 3, 0-52.0 points), human immunodeficiency virus infection (grade 4, 0-48.5 points), retreated tuberculosis history (grade 5, 0-40.0 points), non-standardized treatment regimens of retreated tuberculosis (grade 6, 0-32.5 points), duration of pulmonary cavities (grade 7, 0-31.0 points), passive mode of tuberculosis case finding (grade 8, 0-25.0 points), age<60 years (grade 9, 0-17.5 points), and standard frequencies of chest X-ray examination (grade 10, 0-14.0 points). The C-indexes of this nomogram for the training and validation sets were 0.833 (95% confidence interval ( CI) 0.807-0.859) and 0.871 (95% CI 0.773-0.969), respectively, indicating that the nomogram had good fitting effect. The calibration curves for the risk of incident MDR-TB showed an optimal agreement between nomogram prediction and actual observation in the training and validation sets, respectively.The areas under ROC curve of the 1-year, 5-year, and 10-year MDR-TB risk probability of the training set were 0.904, 0.921, and 0.908, respectively, and those of the validation set were 0.954, 0.970, and 0.919, respectively. Conclusion:Through this nomogram model, clinicians could precisely predict the risk of incident MDR-TB among individuals with PTBH in the clinical practice.

ObjectiveTo understand the experience， knowledge and attitudes of e-cigarette users in Shanghai， and to provide evidence for the development of intervention programs for e-cigarette use. MethodsThe Respondent-Driven Sampling （RDS） method was used to recruit 40 e-cigarette users for qualitative interviews from November 2020 to February 2021. NVivo （11.0） software was used to code interview records， and the thematic pattern and relationship analysis were conducted. ResultsIn terms of the reasons and user experience of e-cigarette use， most respondents used e-cigarettes for the first time after recommendation of friends； The reasons for the current use of e-cigarettes focus on the subjective feelings generated by the use of e-cigarettes （easy to use， replacing traditional tobacco， and having many flavors）. Compared with traditional tobacco use， respondents said e-cigarettes tasted good and had no physical impact （they could be used all the time without making them uncomfortable）. In terms of the awareness of e-cigarette use related knowledge， the respondents' opinions on whether e-cigarettes caused health risks were mostly based on their own experience of using them， and they said it was difficult to identify real information online. As for whether they were interested in relevant knowledge， most respondents said they wanted to learn about the professional knowledge related to e-cigarettes. Regarding the attitude related to the use of e-cigarettes， most respondents believed that e-cigarettes are addictive， e-cigarettes are not a safe alternative to traditional tobacco， ande-cigarettes couldn't help them quit smoking. Three out of ten respondents said they supported including e-cigarettes in the Regulation on Smoking Control in Public. ConclusionThere are certain characteristics of e-cigarette users' feelings， related knowledge and attitude towards e-cigarette use that should be targeted to carry out publicity， education and intervention.

Objective To investigate the effect of voluntary wheel running on negative affective of mice induced by formaldehyde. Methods Thirty male Kunming mice were randomly divided into three groups, including normal saline control group (NS), formaldehyde model group (F), and voluntary wheel running with formaldehyde injection group ( R+ F). The pain model was established by right hindpaw intraplantar formalin injection, the mice of R+F group experienced voluntary wheel running for three weeks before intraplantar formaldehyde injection. The spontaneous pain behavior was determined by the cumulative time of licking paw. The anxiety-like behavior of each group was determined by open field test (OFT) and elevated plus-maze test (EPM) while the depression-like behavior of each group was determined by forced swimming test (FST). The expression of doublecortin ( DCX ) in the hippocampus was determined by immunohistochemistry. Results Compared with the NS group, the typical two-phase pain response was observed in the F group, and compared with the F group, the second phase pain duration was significantly reduced in the R+F group (P<0. 01). In the open field test, the F group showed remarkably reduced time in the inner area(P<0. 001) compared with the NS group, while the R+F group increased time in the inner area (P<0. 05) compared with the F group. In the elevated plus-maze test, the F group showed remarkably reduced time (P< 0. 001) spent in the open arm compared with the NS group, however, compared with the F group, R+F group increased time spent in the open arm (P<0. 05). In the forced swimming test, the immobility time of the F group significant increased (P<0. 01) compared with the NS group, which was decreased in the R+F group (P<0. 05). The Immunohistochemistry showed that the area of DCX positive cells in the hippocampus of the F group was downregulated compared with the NS group, which was upregulated in the R+F group. Conclusion Our findings indicate that voluntary wheel running can improve anxiety and depression-like in mice induced by formaldehyde injection, which may be related to enhanced neurogenesis in the hippocampus.

Objective: To explore the mechanism of reactive oxygen species/thioredoxin-interacting protein/nucleotide-binding oligomerization domain-like receptor 3 (ROS/TXNIP/NLRP3) pathway in the skin injury of trichloroethylene (TCE) sensitized mice. Methods: In August 2020, 40 female BALB/c mice were randomly divided into control group (n=5) , solvent control group (n=5) , TCE treatment group (n=15) and TCE+(2-(2, 2, 6, 6-Tetrameyhylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) triphenylphosphonium chloride (Mito TEMPO) treatment group (n=15) . The TCE sensitization model was established. Mice in the TCE treatment group and TCE+Mito TEMPO treatment group were divided into the sensitized positive group and the sensitized negative group according to the skin erythema and edema reactions on the back of the mice 24 h after the last stimulation. The mice were sacrificed 72 h after the last stimulation, the back skin of the mice was taken, and the skin lesions were observed. Immunohistochemistry (IHC) was used to detect the expression level of NLRP3, and the Western Blot was performed to detect the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) , cysteinyl aspartate specific proteinase 1 (Caspase 1) , Interleukin-1β (IL-1β) and TXNIP proteins in the skin of the mice, the reactive oxygen species (ROS) kit was used to detect the level of intracellular ROS in the back skin tissue. Results: The sensitization rates of TCE treatment group and TCE+Mito TEMPO treatment group were 40.0% (6/15) and 33.3% (5/15) , respectively, and there was no significant difference between the two groups (P>0.05) . The back skin of the mice in the TCE sensitized positive group was thickened and infiltrated by a large number of inflammatory cells. The number of mitochondria in the epidermis cells was significantly reduced, the mitochondrial crest disappeared and vacuolar degeneration occurred. TCE+Mito TEMPO sensitized positive group had less damage, more mitochondria and relatively normal cell structure. Compared with the solvent control group and corresponding sensitized negative groups, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE sensitized positive group and TCE+Mito TEMPO sensitized positive group were significantly increased (P<0.05) . Compared with TCE sensitized positive group, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE+Mito TEMPO sensitized positive group were significantly decreased (P<0.05) . Conclusion: ROS/TXNIP/NLRP3 pathway was activated and then encouraged the release of IL-1β, finally aggravated the TCE-induced skin injury.
Animals ; Carrier Proteins ; Caspase 1/metabolism* ; Female ; Inflammasomes/metabolism* ; Mice ; Mice, Inbred BALB C ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Reactive Oxygen Species/metabolism* ; Solvents ; Thioredoxins/metabolism* ; Trichloroethylene/toxicity*

Objective: “Same treatment for different diseases” is a unique treatment strategy in traditional Chinese medicine. Two kinds of malignant respiratory diseases endanger human health-chronic obstructive pulmonary disease (COPD) and lung cancer. Citrus Grandis Exocarpium (Huajuhong in Chinese, HJH), a famous herbal, is always applied by Chinese medicine practitioners to dispersion the lung to resolve phlegm based on “syndrome differentiation and treatment” theory. However, the common mechanism for HJH's treatment of COPD and lung cancer is not clear. Methods: In this study, based on network pharmacology and molecular docking technology, the common mechanism of HJH in the treatment of COPD and lung cancer was studied. The active ingredients and related targets of HJH were integrated from TCMSP, BATMAN-TAM, STP, and Pubchem databases. The standard names of these targets were united by UniProt database. Targets of COPD and lung cancer were enriched through GeneCards, NCBI (Gene), Therapeutic Target Database, and DisGeNET (v7.0) databases. Then the intersection targets of HJH and diseases were obtained. The STRING network and the Cytoscape 3.7.2 were used to construct PPI network, the DAVID database was used to perform GO and KEGG analysis. Then Cytoscape 3.6.1 was used to build “ingredient-target-signal pathway” network. Finally, AutoDock 1.5.6 software was used to perform molecular docking of key proteins and molecules. Results: Eleven active ingredients in HJH were obtained by searching the database, corresponding to 184 HJH-COPD-lung cancer targets intersection. The results of biological network analysis showed that naringenin, the active component in HJH, could mainly act on target proteins such as AKT1, EGFR. Then through positive regulation of vasoconstriction and other biological processes, naringenin could regulate estrogen signaling pathway, VEGF signaling pathway, HIF-1 signaling pathway, ErbB signaling pathway, PI3K-Akt signaling pathway to play an important role in the treatment of both COPD and lung cancer. Conclusion: Network pharmacology was employed to systematically investigate the active ingredients and targets of HJH in treatment of COPD and lung cancer. And then, the common pharmacodynamic network of HJH for the two malignant respiratory diseases was firstly described. Furthermore, naringenin was proved to strongly bind with AKT1 and EGFR. It may provide the scientific basis for understanding the “Same treatment for different diseases” strategy in traditional Chinese medicine and inspirit subsequent drug discovery for COPD, lung cancer and other malignant lung diseases.