Objective To explore the role of conventional protein kinase C(cPKCs) in delayed hypoxic preconditioning.Methods The biochemistry techniques of SDS-PAGE,Western bolt and Gel Doc imagine were applied to analyze the effect of repetitive hypoxic exposure(H5,H6) on the level of cPKC?,? membrane translocation and protein expression in murine brain.Results We found that cPKC? protein expression was significantly decreased(P

Objective To evaluate the role of spinal sigma-1 receptors in the maintenance of bone cancer pain (BCP) in rats and the relationship with extracellular signal-regulated kinase (ERK).Methods Part Ⅰ Twenty-four female Sprague-Dawley rats,weighing 180-220 g,were randomized into 2 groups using a random number table:sham operation group (S group,n =4) and BCP group (n =20).BCP was induced by inoculating Walker 256 mammary gland carcinoma cells into the medullary cavity of the right tibia.Four rats were sacrificed on day 10 after inoculation in S group or on day 3,5,7,10 and 14 after inoculation in BCP group,and the L4-6 segments of the spinal cord were removed to measure the expression of sigma-1 receptors by Western blot.Part Ⅱ Forty female Sprague-Dawley rats,weighing 180-220 g,were randomized into 4 groups (n =10 each) using a random number table:sham operation group (S group),sigma-1 receptor inhibitor BD1047 group (BD group),BCP group,and BCP + BD1047 group (BCP + BD group).On day 10 to 14 after inoculation,normal saline 20 μl was injected intrathecally once a day in S and BCP groups,or BD1047 120 nmol/20μl was injected intrathecally once a day in BD and BCP + BD groups.Mechanical paw withdrawal threshold (MWT) to yon Frey filament stimulation was measured one day before inoculation,on day 3,5 and 7 after inoculation,and on day 10,12 and 14 after administration.After measurement of MWT on day 14 after inoculation,the rats were sacrificed and the L4-6 segments of the spinal cord were removed to determine the expression of phosphorylated ERK (p-ERK) by Western blot.Results Part Ⅰ Compared with group S,the expression of sigma-1 was significantly up-regulated and peaked on day 10 after operation in group BCP.Part Ⅱ Compared with S group,no significant changes were found in MWT and p-ERK expression at each time point in BD group,and MWT was decreased and p-ERK expression was up-regulated in BCP and BCP + BD groups.Compared with group BCP,after intrathecal injection of BD1047,MWT was significantly increased and the expression of p-ERK was down-regulated in BCP + BD group.Conclusion Spinal sigma-1 receptors are involved in the maintenance of BCP in rats possibly through promoting phosphorylation of ERK.

BACKGROUND: The protein kinase C (PKC) family consists of 3 groups of PKCs, namely the conventional PKC (cPKC), atypical PKC and novel PKC.Accumulating studies conducted in recent years have suggested that PKCs may play important roles in the development of cerebral ischemic/hypoxic preconditioning ( I / HPC ).OBJECTIVE: To observe membrane translocation of hypoxia-activated cPKC isoforms(α, βⅠ, βⅡ and γ) at cellular levels in a cell hypoxia model.DESIGN: Randomized block design.SETTING: Department of Neurobiology, College of Basic Medical Sciences,Capital University of Medical Sciences.MATERIALS: The experiment was completed at the Neurobiological Cell Culture Laboratory of Capital University of Medical Sciences in May 2004.Human neuroblastoma cells with the properties of neurons were maintained and passaged in this laboratory.METHODS: The activation of cPKC isoforms under hypoxic condition and changes of cPKCα, βⅠ, βⅡ and γ membrane translocation(an indicator of PKCs activation) in SH-SY5Y neuroblastoma cells in response to hypoxia (1% 02, 5% CO2 and 94% N2) for 0 to 24 hours were observed using SDS-PAGE, Western blotting and immunocytochemistry.MAIN OUTCOME MEASURES: Effect of sustained hypoxia on cPKC membrane translocation in human neuroblastoma cells was observed with SDS-PAGE, cPKC Western blotting, and immunocytochemistry.RESULTS: cPKCα, βⅠ and βⅡ membrane translocation were increased significantly ( P ＜ 0.05 ) in a time-dependent manner in response to hypoxic exposure, and the increase of cPKCβⅠ was more evident( P ＜ 0. 001 ) after 4hours of hypoxic exposure, whereas no cPKCγ was detected in SH-SY5Y neuroblastoma cells either under normoxic or hypoxic condition. The results suggested that all cPKC isoforms, epically cPKCβⅠ, could be activated by sustained hypoxia, and the absence of cPKCγ in SH-SY5Y neuroblastoma cells may be relevant to the loss of specific biological features of the cultured cells.CONCLUSION: Sustained hypoxia activates the isoforms of cPKCα, βⅠ and βⅡ in human neuroblastoma cells and induces their membrane translocation.cPKCγ isoform may not exist in human neuroblastoma cells, or the cells has lost certain biological characteristics.

Objective: To investigate the household eye hygiene among students at grades 3 to 5 in primary schools in Jinshan District, Shanghai Municipality, so as to provide insights into the management of myopia among children and adolescents. Methods: Students at grades 3 to 5 in primary schools were sampled in Jinshan District using a stratified cluster sampling method from December 2021 to January 2022. The household reading environments were observed, including the height of the learning desk and chair, desk surface color and strength. The duration of reading and writing, duration of watching TV and videos, duration of outdoor activities, duration of sleep and eye use behaviors and habits after school and at weekends were investigated using questionnaires. Results: A total of 330 primary school students were surveyed, including 179 boys (54.24%) and 151 girls (45.76%), and there were 36.36% grade 3 primary school students, 36.36% grade 4 students and 27.27% grade 5 students. There were 94.24% of primary school students that used the desk and chair with heights mismatched to students' heights, 25.45% that used desks with dark surface, 26.67% that used desks with light reflection, and 48.48% that used desk lamps with an illumination intensity of <300 lx. There were 56.36% of students with reading and writing duration of 1 h and longer, 15.76% with watching duration of 0.5 h and longer, 86.97% with outdoor activity duration of <1 h and 88.48% with sleep duration of <10 h after school, and 42.42% with reading and writing duration of 2 h and longer, 29.70% with watching duration of 1 hour and longer, 65.45% with outdoor activity duration of <2 h and 55.76% with sleep duration of <10 h at weekends. There were 30.91% of primary school students with 10 cm distance from chest to desk, 26.36% with 33 cm distance from eyes to books and 35.15% with 3 cm distance from fingers to pen points when reading and writing, and 35.45% with >3 m distance from TV, 40.91% with >50 cm distance from computers, and 22.73% with >40 cm distance from cell phones when watching TV or videos. Conclusions The household eye use environments remain to be improved, and there are poor eye use behaviors among primary school students at grades 3 to 5 in Jinshan District, Shanghai Municipality; notably, mismatch between the desk and chair height and students' body height and inadequate sleep are common.

Objective : To analyze the epidemiological characteristics of food-borne diseases in Jinshan District, Shanghai from 2014 to 2020, so as to provide the evidence for formulating the public health strategy for food-borne diseases control. Methods : The medical records of patients with food-borne diseases were collected from 16 monitoring hospitals in Jinshan District from 2014 to 2020, and the basic information, clinical symptoms, history of suspicious dietary exposure and disease diagnosis were extracted. The crowd distribution, temporal distribution, spatial distribution, history of suspected dietary exposure and etiological characteristics of patients with food-borne diseases were descriptively analyzed. Results : A total of 1 060 cases with food-borne diseases were reported in Jinshan District from 2014 to 2020, including 1 057 cases with infectious diseases ( 99.72% ) and 3 poisoning cases ( 0.28% ). The male/female ratio of the cases was 0.94∶1, and 47.55% ( 504 cases ) were at ages of 15 to 44 years. Working ( 402 cases, 37.92% ) and farming ( 218 cases, 20.57% ) were predominant occupations, and the detection of food-borne diseases was concentrated between May and October, with two peaks seen in August and May. The suspicious food exposure was predominantly meat and meat products (215 cases, 20.28%), and the suspicious food exposure place was predominantly at home ( 363 cases, 34.25% ). In addition, the positive rate of food-borne infection was 23.03% in 712 samples, including 123 samples with Vibrio parahaemolyticus infections ( 17.83% positive rate ), and 26 samples with Salmonella infections ( 3.65% positive rate ). Conclusion Food-borne diseases were highly prevalent in summer in Jinshan District from 2014 to 2020, and infectious cases were predominant. Young people, workers and farmers are at high risk of food-borne diseases, and Vibrio parahaemolyticus and Salmonella were predominant pathogens.

This study was aimed to explore the effect of Periplaneta Americana against alcohol-induced liver injury in mice, in order to provide a theoretical basis for the industrial production of P. Americana. Kunming mice were randomly divided into six groups, which were the normal group, model group, positive group, high-, middle-, low-dose of P. Americana groups. Intragastric administration of Tiopronin Enteric-coated Tablets 100 mg·kg-1 was given to the positive group. Intragastric administrations of whole powder of medicine were given to the high-, middle-, low-dose groups with the dosage of 6.667, 3.333, 1.667 g·kg-1, respectively. The drugs were given daily for 10 con-secutive days. After 3h of the 10th day drug administration, intragastric administration of distilled water was given to the normal group, while 14mL·kg-1 of 56℃ Red Star Liquor was given to other groups. No food was given but water for 12h. Blood was collected from the orbit. The ALT, AST and GGT in blood serum of mice were measured. The liver was dissected and liver coefficient was calculated. Histopathological examination was given on liver tissues. The results showed that compared with the normal group, the level of ALT, AST in blood serum of the model group had obvious enhanced (P< 0.01), the level of GGT had obvious enhanced (P< 0.05). Compared with the model group, the level of GGT, AST of the high-, middle-dose group had obvious enhanced (P< 0.05), the level of ALT activity had obvious enhanced (P< 0.01). There were severe liver histopathological damages in mice of the high-, middle-, low-dose group. It was concluded that P. Americana had some side effects in the treatment of alcohol-induced liver injury.

Objective:To investigate the predictive value of serum C-type lectin-like receptor 2 (CLEC-2) combined with insulin resistance in the outcome of patients with acute ischemic stroke (AIS) after intravenous thrombolysis.Methods:Patients with AIS received alteplase intravenous thrombolytic therapy in the Department of Neurology, the Second Affiliated Hospital of Nantong University from October 2019 to March 2021 were enrolled retrospectively. According to the modified Rankin Scale score at 90 d after onset, they were divided into good outcome group (0-2) and poor outcome group (>2). Homeostasis model assessment of insulin resistance (HOMA-IR) was used to evaluate insulin resistance. Person correlation analysis was used to determine the correlation between CLEC-2 and HOMA-IR. Multivariate logistic regression analysis was used to determine the correlation between serum CELC-2, HOMA-IR and the outcome after intravenous thrombolysis. Receiver operating characteristic (ROC) curve was used to determine the predictive value of serum CLEC-2 combined with HOMA-IR for poor outcome after intravenous thrombolysis. Results:A total of 100 patients were enrolled (56 males, 56.0%; aged 70.6±10.86 years, range 49-83 years). The baseline National Institutes of Health Stroke Scale (NIHSS) score was 10.00±6.36. Senenty-four patients (74.0%) had a good outcome and 26 (26.0%) had a poor outcome. Person correlation analysis showed that there was a significant positive correlation between serum CLEC-2 and HOMA-IR ( r=0.523; P<0.001). Multivariate logistic regression analysis showed that after adjusting for confounding factors (C-reactive protein, baseline NIHSS score, onset-to-needle time), the highest quartile of serum CLEC-2 (compared with the lowest quartile: odds ratio [ OR] 4.836, 95% confidence interval [ CI] 1.105-21.169; P=0.036) and the highest quartile of HOMA-IR (compared with the quartile 1-3: OR 15, 95% CI 2.647-30.722; P=0.002) were the independent risk factors for the poor outcome in patients with AIS after intravenous thrombolysis. ROC curve analysis showed that the area under the curve for serum CLEC-2 combined with HOMA-IR to predict poor outcome was 0.785 (95% CI 0.688-0.883; P<0.001), the optimal cut-off value was 0.72, and the sensitivity and specificity were 76.0% and 95.0%, respectively. Conclusion:CLEC-2 combined with insulin resistance has a certain predictive value for the poor outcome of patients with AIS after intravenous thrombolysis.

Objective:To investigate the effect of oleanolic acid (OA) on Sirtuin1 (SIRT1)-mediated high-mobility group box 1(HMGB1) deacetylation in the early brain injury after subarachnoid hemorrhage (SAH).Methods:A total of 176 male Sprague-Dawley rats were randomly divided into Sham operation (Sham group) ( n=48), SAH group ( n=48), OA group ( n=48) and Sirtinol group ( n=32). Rats in the SAH group, OA group and the Sirtinol group all adopted internal carotid artery puncture to construct SAH model, while rats in the sham group did not adopt puncture. One hour after modeling, the rats in the OA group were given intraperitoneal injection of OA (20 mg/kg), and the rats in the Sirtinol group were given intracerebroventricular injection of Sirtinol (2 mmol/L, 30 μL/kg). The rats in the sham group and SAH group were injected with equal volumes of sodium chloride injection. The SAH score and neurological score were performed 24 h after SAH, and the water content in the brain tissue and Evans blue exudation rate were measured. The expressions of HMGB1, SIRT1 and acetylated HMGB1 proteins in the brain tissue of rats were detected by Western Blot. The expression of HMGB1 mRNA in the brain of the rats was detected by quantitative real-time PCR. The distribution of HMGB1 protein in the brain of the rats was observed by immunofluorescence staining. TUNEL staining was used to observe the neuronal apoptosis in the brain tissue of the rats. Results:Compared with the SAH group, the SAH score of the OA group was significantly reduced ( P<0.001), the Garcia score was increased ( P<0.01), and the brain water content and Evans blue exudation rate were both reduced (all P<0.01). Compared with the OA group, the SAH score of the Sirtinol group was increased ( P<0.01), the Garcia score was significantly decreased ( P<0.001), and the brain water content and Evans blue exudation rate were both increased (all P<0.01). The results of Western Blot and real-time fluorescent quantitative PCR showed that, compared with the SAH group, the protein level ( P<0.01) and mRNA level ( P<0.05) of HMGB1 in the OA group were decreased, the expression of SIRT1 protein was significantly increased ( P<0.001), and the expression of acetylated HMGB1 protein was decreased ( P<0.01). Immunofluorescence staining showed that OA inhibited the migration of HMGB1 protein from the nucleus to the cytoplasm. TUNEL staining showed that OA could effectively reduce the number of TUNEL-positive cells. Compared with the OA group, Sirtinol significantly increased the number of TUNEL-positive cells. Conclusions:OA can reduce the release of HMGB1 through the SIRT1/HMGB1 pathway, thereby protecting the early brain injury after SAH.

Objective:To discuss the antioxidant and anti-apoptosis effects of 3-hydroxy-olea- 12-en-28-oic acid (OA) on subarachnoid hemorrhage (SAH) and their mechanisms in rat models.Methods:In experiment one: 144 SD rats were divided into sham-operated group, SAH group and SAH+20 mg/kg OA group by random number table method ( n=48); SAH models were established by standard endovascular puncture method; rats in the SAH+20 mg/kg OA group received intraperitoneal injection of OA one h after modeling, while rats in the sham-operated group received the same operation without perforation; brain edema, destruction of blood-brain barrier, SAH severity degrees (Sugawa scale scores), neurological function scale scores and other early brain injury-related indicators were detected in rats from the three groups; the levels of malondialdehyde (MDA), superoxide dismutase (SOD) were investigated; the activation of nuclear factor related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) axis and neuronal apoptosis after SAH were detected. In experiment two: another 36 rats were divided into sham-operated group, SAH model group, sham operated+negative control siRNA group, SAH+negative control siRNA group, sham operated+Nrf siRNA group and SAH+Nrf siRNA group according to random number table method ( n=6); treatment methods in the sham-operated group and SAH modeling methods were the same as experiment one; negative control siRNA or Nrf siRNA were injected into the lateral ventricle of rats in the later 4 groups 24 h before SAH modeling; expressions of apoptosis-related factors in the downstream of Nrf2/HO-1 axis were detected in rats from the 6 groups. Results:In experiment one: (1) as compared with rats in the SAH group, rats in the SAH+20 mg/kg OA group had significantly decreased Sugawa scale scores, statistically increased neurological function scale scores and balance beam scale scores, significantly decreased brain moisture content and Evans blue exhalation rate ( P<0.05); (2) as compared with SAH group, SAH+20 mg/kg OA group had significantly decreased MDA level, and significantly increased SOD, glutathione peroxudase and catalase levels ( P<0.05); (3) the expressions of Nrf2, HO-1 and B-cell lymphoma-2(Bcl-2) were significantly increased, the expressions of apoptosis-related factors cytochrome-C, caspase-3 and activated caspase 3 were signficantly decreased, and the number of apoptotic neurons was significantly samller in the SAH+20 mg/kg OA group as compared with those in the SAH group ( P<0.05). In experiment two: as compared with those in the SAH+negative control siRNA group, the protein expressions of HO-1 and Bcl-2 in the SAH+Nrf siRNA group were significantly decreased, while the expression levels of cytochrome C, caspase-3 and activated caspase 3 were statistically increased ( P<0.05). Conclusion:OA can reduce the occurrence of oxidative stress and neuronal apoptosis by activating Nrf2/HO-1 axis.

Objective:To investigate the effect of rutin on blood-brain barrier in early brain injury after subarachnoid hemorrhage (SAH) in rats.Methods:A rat model of SAH was induced by puncturing the internal carotid artery. The rats were divided into a sham operation group, a model group and a rutin (50 mg/kg) group. Twenty-four hours after modeling, SAH score and neurological deficit score were conducted, and brain water content and Evans blue extravasation rate were detected in each group. Western blot analysis was used to detect the expression of claudin-5, occludin and zonula occluden (ZO)-1. TUNEL staining was used to detect neuronal apoptosis.Results:Compared with the sham operation group, the SAH score increased, the neurological deficit score decreased, and the brain water content and Evans blue extravasation increased in the model group. Rutin could significantly reduce the SAH score, increase neurological deficit score, and reduce brain water content and Evans blue exudation (all P<0.01). Western blot analysis showed that the expression of claudin-5, occludin and ZO-1 protein decreased in the model group, and the expression of claudin-5, occludin and ZO-1 protein increased significantly in the rutin group ( P<0.01). In addition, the number of TUNEL positive cells induced by SAH in the rutin group decreased. Conclusion:Rutin can play a protective role in early brain injury after SAH, and its mechanism may be associated with protecting the integrity of blood-brain barrier.