Aim To observe the effects of theaflavin on intracellular calcium concentration(i) level in rat ventricular myocytes and discuss the possible mechanisms.Methods The effects of theaflavin oni were investigated in rat ventricular myocytes.i was detected by laser confocal microscopy and represented by relative fluorescent intensity(FI-FI0)/FI0,%;FI0:control;FI:administration of drugs).Results ① Theaflavin(10,20,40 ?mol?L-1) had no effect on the i of ventricular myocytes in normal Tyrode′s solution.However,it reduced the i of ventricular myocytes in simulated ischemia solution in a concentration-dependent manner.② Pretreatment with Bay k8644(0.5 ?mol?L-1) mostly abolished the effects of theaflavin(20 ?mol?L-1) in simulated ischemia solution.③ Theaflavin(20 ?mol?L-1) markedly inhibited the low concentration of ryanodine-induced i increase in Ca2+-free Tyrode′s solution.④ When the propagating waves of elevated i(Ca2+ waves) were produced by increasing extracellular Ca2+ concentration from 1 mmol?L-1to 10 mmol?L-1,theaflavin(20 ?mol?L-1) could block the propagating waves of elevated i(Ca2+ waves),reduce the frequency and duration of propagating waves,and reduce i as well.Conclusion Theaflavin may reduce the i in isolated rat ventricular myocytes via inhibiting Ca2+ influx by voltage-dependent Ca2+ channel and alleviating Ca2+ release from sarcoplasmic reticulum(SR).

Objective To investigate the activation state of and expressions of surface co-stimulatory molecules on peripheral CD4+ T cells from patients with pemphigus and healthy human controls.Methods Ninety patients with pemphigus including 24 patients with first-onset pemphigus,51 with quiescent pemphigus and 15 with recurrent pemphigus,as well as 30 healthy human controls were enrolled in this study.Peripheral blood samples were obtained from these subjects followed by lymphocyte isolation.Flow cytometry was performed to detect the expressions of CD69,intercellular adhesion molecule-1 (ICAM-1),inducible co-stimulatory molecule (ICOS),CD40 ligand (CD40L) and OX40 on CD4+ T cells.Statistical analysis was done by Mann-Whitney test using Graphpad 5.0 software.Results The expression rate of CD69 on peripheral CD4+ T cells from the healthy human controls was significantly lower than that from patients with pemphigus,patients with first-onset pemphigus,patients with quiescent pemphigus,and patients with recurrent pemphigus ((1.26 ± 0.19)％ vs.(2.46 ± 0.19)％,(2.77 ± 0.40)％,(2.15 ± 0.25)％ and (2.36 ± 0.35)％,all P ＜ 0.05).The patients with pemphigus also showed a significant increase in the expression rates of ICAM-1,CD40L and OX40 compared with the healthy human controls ((55.88 ± 1.67)％ vs.(47.75 ± 2.52)％,P＜ 0.05; (2.23 ± 0.22)％ vs.(0.73 ± 0.07)％,P＜ 0.01; (2.55 ± 0.29)％vs.(0.62 ± 0.17)％,P ＜ 0.01).No significant differences were observed between patients with different stages of pemphigus in the expression rates of CD69,ICAM-1,CD40L or OX40 (all P ＞ 0.05).The percentage of ICOS-expressing CD4+ T cells was significantly up-regulated in only patients with first-onset pemphigus as compared to the healthy controls ((3.73 ± 0.60)％ vs.(2.39 ± 0.16)％,P ＜ 0.05).Conclusions The peripheral blood CD4+ T cells from patients with pemphigus are in a relatively active state with up-regulated surface expressions of many costimulatory molecules,suggesting that CD4+ T cells are involved in the initiation and progression of pemphigus by interacting with B cells through co-stimulatory molecules.

Objective To explore the effect of acupuncture-rehabilitation therapy on neurological function and expression of Flt-1 and Flk-1, members of vascular endothelial growth factor receptors, after permanent focal cerebral ischemia in rats. Methods Ninety male Sprague-Dawley rats were divided into five groups, namely sham group, model group, acupuncture group, rehabilitation group and acupunc-ture-rehabilitation group, and each group was further divided into 3-day, 7-day and 14-day subgroups, equally. Their middle cerebral arteries were occluded except those of sham group. The sham and model groups accepted no treatment, while the acupuncture group accepted clus-ter needling of scalp acupuncture, the rehabilitation group accepted treadmill training, and the acupuncture-rehabilitation group accepted both acupuncture and treadmill training. They were assessed with modified Neurologic Severity Score (mNSS) 3, 7 and 14 days after model-ing, while the expression of Flt-1 and Flk-1 were determined with Western blotting. Results The mNSS score reduced in all the treatment groups (P<0.05) compared with that of the model group at every time point, and was the least in the acupuncture-rehabilitation group (P<0.05) 7 and 14 days after modeling among the treatment groups. Meanwhile, the expression of Flt-1 and Flk-1 protein increased in all the treatment groups (P<0.05), and was the most in the acupuncture-rehabilitation group (P<0.05). Conclusion Acupuncture-rehabilitation thera-py can promote the neurological function recovery in rat with permanent focal cerebral ischemia, which may be associated with the continu-ous inducement of Flt-1, Flk-1 protein expression in ischemic penumbra cortex.

Objective: To explore the epidemiological characteristics of mortality of accidental injury children aged 5-19 years in Yunnan Province during 2015-2019, to provide further evidence for reducing accidental mortality in children. Methods: The mortality cases of accidental injury by gender, age groups, and causes among children and adolescents in Yunnan Province during 2015 to 2019 were analyzed retrospectively. Poisson regression model was used to estimate the trend of mortality. Chi square test was used to compare the differences of mortality by gender and age groups. Results: During 2015 to 2019, the mortality rate of accidental injury in children and adolescents aged 5-19 years in Yunnan Province decreased from 19.15/10 5 in 2015 to 18.35/10 5 in 2019 (Z=-3.36, P <0.01). The mortality rate of all types of accidental injuries in male was higher than that in female ( χ 2=867.01, 11.69 , 147.60, 190.34, 7.23, 702.97, P <0.05). The top three causes of fatal accidental injuries included road traffic accidents, drowning and falls. Except for accidental falls, the mortality rate of other causes in the 15-19 age group was significantly higher than that in the age of 5-9 years and 10-14 years groups ( χ 2=764.47, 75.91, 31.75, 9.24, 114.96, 327.64, P <0.05). The top three traffic accidents were happened among motorcyclists, light trucks or caravans and cars, accounting for 27.83%, 10.57% and 7.90% respectively. Conclusion Targeted measures should be taken continuously according to the characteristics of children and adolescents in different regions, age groups and gender to reduce the incidence and mortality of accidental injuries among children.

Bullous pemphigoid (BP) is the most common autoimmune subepidermal blistering skin disease,and a large number of eosinophils (EOS) obviously infiltrate the dermis of BP lesions.Additionally,EOS and their activated cytokines and chemokines are abundantly present in blisters and peripheral blood of BP patients.It is also proved that EOS can induce the dermal-epidermal separation.The level of interleukin (IL)-5 is markedly increased in the sera and blister fluids of BP patients,and IL-5 secreted by Th2 cells and EOS can regulate the differentiation,activation and survival of EOS.All the above evidence indicates that EOS are associated with the occurrence of BP.Researches on EOS and their cytokines will develop a new field of targeted therapy for BP.

In daily medical work, most of the critically ill patients who cannot move by themselves are pulled and lifted by manpower, often relying on the cooperation of many doctors and nurses, which not only increases the risk of transfer and patients' discomfort, but also causes certain skeletal and muscle damage to the porters. The emergency department of the First Hospital of Jiaxing City, Zhejiang Province designed a kind of patient transfer device, and obtained the National Utility Model Patent (ZL 2018 2 0579844.X). The transfer device is composed of upper frame, lower frame and base. The upper frame and the lower frame are rectangular and in a horizontal position. The upper frame can slide laterally through the circular tubes which are fixed on the lower frame. The lower part of the base is provided with four universal foot brake wheels. During the usage, the booster frame facilitates the transfer of patients by the rolling and two sliding tracks of the circular tube, which can make patients move smoothly and comfortably, and reduce the working intensity of the transporter. This device has good practical value.

Objective:To evaluate the relationship between hippocampal miR-3065-5p and insulin-like growth factor-1/phosphatidylinositol 3-kinase/protein kinase B(IGF-1/PI3K/Akt)signaling pathway in a mouse model of perioperative neurocognitive disorder (PND).Methods:Eighty clean-grade healthy male C75BL/6 mice, aged 12-14 weeks, weighing 20-30 g, were divided them into 4 groups ( n=20 each) using the random number table method: control group (C group), PND group, miR-3065-5p agonist group (Ag group) and miR-3065-5p agonist negative control group (Ag-NC group). PND model was prepared by internal fixation of tibial fracture under anesthesia with 1.5% isoflurane. Two days before developing the model, miR-3065-5p agomir 2 μl was injected into the lateral ventricle in Ag group, miR-3065-5p agomir negative control 2 μl was injected into the lateral ventricle in Ag-NC group. Morris water maze test and open field test were performed at 7 days after surgery. The mice were sacrificed after the end of test, and hippocampal tissues were obtained for determination of the expression of miR-3065-5p, IGF-1 mRNA and Bcl-2 mRNA (by quantitative real-time polymerase chain reaction) and expression of IGF-1, phosphorylated Akt (p-Akt), phosphorylated glycogen synthase kinase-3β (p-GSK3β) and Bcl-2 (by Western blot). Results:There was no significant difference in each parameter in the open field test among the four groups ( P>0.05). Compared with group C, the postoperative escape latency was significantly prolonged, the percentage of time of stay at the target quadrant was decreased, the number of crossing the original platform was reduced, the expression of miR-3065-5p was up-regulated, and the expression of IGF-1 mRNA, Bcl-2 mRNA, IGF-1, p-Akt, p-GSK3β and Bcl-2 was down-regulated in the other three groups ( P<0.05). Compared with PND group and Ag-NC group, the postoperative escape latency was significantly prolonged, the percentage of time of stay at the target quadrant was decreased, the number of crossing the original platform was reduced, the expression of miR-3065-5p was up-regulated, and the expression of IGF-1 mRNA, Bcl-2 mRNA, IGF-1, p-Akt, p-GSK3β and Bcl-2 was down-regulated in Ag group ( P<0.05). Conclusions:Up-regulation of miR-3065-5p can inhibit the activation of IGF-1/PI3K/Akt signaling pathway, which might be one of the mechanisms of PND developed in mice.

Objective:To investigate the effects of mild hypothermia on microglia polarization and janus kinase 2/signal transduction and transcriptional activation factor 3 (JAK2/STAT3) signaling pathway during cerebral ischemia-reperfusion (I/R) in rats.Methods:Forty-five clean-grade healthy male Sprague-Dawley rats, aged 8 weeks, weighing 260-280 g, were divided into 3 groups ( n=15 each) by the random number table method: sham operation group (S group), cerebral I/R group (I/R) and mild hypothermia group (H group). In I/R group and H group, cerebral I/R was induced by middle cerebral artery occlusion using a nylon thread in anesthetized animals, the nylon thread was removed to restore the perfusion after 2 h of occlusion, and the rectal temperature was maintained at 36-37 ℃ during the period. Group H was wiped with 75% alcohol for 3 h starting from the time point immediately after reperfusion, and the rectal temperature was maintained at 32-33℃. Modified neurological severity score (mNSS) was evaluated at 24 h of reperfusion. Animals were then sacrificed for determination of the cerebral infarct size (using TTC staining), expression of M1 marker inducible nitric oxide synthase (iNOS), M2 marker arginase 1(Arg-1), phosphorylated JAK2(p-JAK2)and phosphorylated STAT3(p-STAT3)(by Western blot), expression of iNOS mRNA and Arg-1 mRNA (by quantitative polymerase chain reaction), and contents of interleukin-6 (IL-6) and IL-10 (by enzyme-linked immunosorbent assay). Results:Compared with group S, mNSS and cerebral infarct size were significantly increased, the expression of iNOS, Arg-1 protein and mRNA in cerebral ischemic penumbral zone was up-regulated, and the p-JAK2/JAK2 ratio, p-STAT3/STAT3 ratio, and contents of IL-6 and IL-10 were increased in the other two groups ( P<0.05). Compared with I/R group, mNSS and cerebral infarct size were significantly decreased, the expression of iNOS protein and mRNA in cerebral ischemic penumbral zone was down-regulated, the expression of Arg-1 and mRNA was up-regulated, and the p-JAK2/JAK2 ratio, p-STAT3/STAT3 ratio and IL-6 content were decreased, and the IL-10 content was increased in group H ( P<0.05). Conclusions:Mild hypothermia can promote the polarization shift of microglia from M1 to M2 phenotype during cerebral I/R and inhibit the central inflammatory responses, and the mechanism may be related to inhibition of JAK2/STAT3 signaling pathway in rats.

Objective:To evaluate the role of glutathione S-transferase μ1 (GSTM1) expression in mild hypothermia-induced mitigation of cerebral ischemia-reperfusion (I/R) injury and the relationship with microglial polarization.Methods:Eighty clean-grade healthy male Sprague-Dawley rats, aged 8 weeks, weighing 260-280 g, were divided into 4 groups ( n=20 each) using a random number table method: sham operation group (S group), cerebral I/R group (I/R group), mild hypothermia group (H group), and GSTM1 inhibitor + mild hypothermia group (IH group). The rat model of cerebral I/R injury was prepared using the filament occlusion method. The filament was removed to restore blood flow after the left middle cerebral artery was blocked for 2 h, and the rats′ brain and rectal temperature were maintained at 36-37 ℃ during the period. The vessels were only isolated and ligated without occlusion in S group. In H group, the entire body was wiped with 75% ethanol immediately after removing the filament, and the brain and rectal temperatures were maintained at 32-33 ℃ for 3 h, and the other procedures were the same as those previously described in I/R group. In IH group, GSTM1 inhibitor itaconic acid 8.6 mg/kg was intraperitoneally injected at 24 and 1 h before developing the model, and the other procedures were the same as those previously described in H group. Neurological deficits were evaluated using a modified neurological severity score (mNSS) at 24 h of reperfusion, and then the animals were sacrificed and the brains were removed for observation of cerebral infarction (by TTC staining) and for determination of the expression of GSTM1, M1-type microglial marker inducible nitric oxide synthase (iNOS), and M2-type microglial marker arginase-1 (Arg-1) (by Western blot), expression of GSTM1, iNOS and Arg-1 mRNA (quantitative real-time polymerase chain reaction) and contents of interleukin-6 (IL-6), IL-10, tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β) (by enzyme-linked immunosorbent assay). Results:Compared with S group, the mNSS and percentage of cerebral infarct size were significantly increased, and the expression of iNOS and Arg-1 protein and mRNA was up-regulated, the expression of GSTM1 and mRNA was down-regulated, and the contents of IL-6, TNF-α, IL-10 and TGF-β were increased in the other three groups ( P<0.05). Compared with I/R group and IH group, the mNSS and percentage of cerebral infarct size were significantly decreased, and the expression of iNOS protein and mRNA was down-regulated, the expression of Arg-1 protein and mRNA and GSTM1 was up-regulated, the contents of TNF-α and IL-6 were decreased, and the contents of TGF-β and IL-10 were increased in H group ( P<0.05). Conclusions:Up-regulated expression of GSTM1 is involved in mild hypothermia-induced mitigation of cerebral I/R injury, which is associated with inhibition of microglial polarization toward the M1 phenotype and promotion of polarization toward the M2 phenotype.

Objective:To explore any effect of repeated transcranial magnetic stimulation (rTMS) on the upper limb motor function and cerebral cortex activation of stroke survivors.Methods:Sixty stroke survivors were randomly divided into an intervention group and a control group, each of 30. In addition to routine rehabilitation training (including drug therapy, comprehensive hemiplegic limb training and physical factor therapy), the intervention group received 15 minutes of rTMS daily, five days a week for 4 weeks while the control group was given false rTMS. Upper limb motor function was evaluated before and after the treatment using the Fugl Meyer upper limb motor function rating scale (FMA-UE). Functional near-infrared spectroscopy was used to detect and compare the activation (β values) of the prefrontal cortex, the motor cortex and the primary somatosensory cortex in the 2 groups. The correlation between the FMA-UE scores and the β values was quantified.Results:①There was no significant difference in the average FMA-UE scores between the two groups before the treatment. Afterward, though both groups′ average scores had increased significantly, there was significantly greater improvement in the treatment group. ②There was also no significant difference in average β value between the two groups before the experiment, but afterward the average βs of channels 27 and 13 in the intervention group were significantly higher than in the control group. Moreover, in patients with lesion in the left brain, the β-values of CH27 and CH13 were also significantly higher than the control group ( P<0.05). ③The FMA-UE scores of the intervention group were moderately correlated with the CH27 and CH13 β values, but those of the control group were only weakly correlated with the β values of CH27. Conclusion:Transcranial magnetic stimulation activates a lesioned left brain region, improving upper limb motor function. The improvement is correlated with the activation of the left prefrontal cortex and the left primary somatosensory cortex.