BACKGROUND:Previous animal studies have shown that riluzole can inhibit neuroinflammatory response after spinal cord injury and promote functional recovery in injured rats,but the study on whether it can regulate the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome in the acute stage is lacking. OBJECTIVE:To observe whether riluzole can reduce microglial pyroptosis and promote functional recovery after spinal cord injury by modulating NLRP3 inflammasome through animal experiments,histological experiments and molecular biology experiments. METHODS:Female SD rats were divided into sham operation,model and riluzole groups,with 12 rats in each group.In addition to the sham operation group,T10 spinal cord injury was conducted in rats.The model group was treated with intraperitoneal administration of riluzole with solvent cyclodextrin.The riluzole group was treated with a 4 mg/kg dose of riluzole injection.The effect of riluzole on motor function recovery was assessed using the BBB score and inclined plane test.The recovery of sensory-evoked potential and motor-evoked potential was measured by electrophysiology.Hematoxylin-eosin staining was used to evaluate spinal cord tissue repair.The regulatory effects of riluzole on NLRP3,Caspase-1 and gasdermin D protein expression in spinal cord tissues were detected by western blot assay.ELISA was utilized to detect the expression levels of inflammatory factors interleukin-1β and interleukin-18.The effects of riluzole on the expression of NLRP3,Caspase-1,gasdermin D and interleukin-1β in microglial cells of the injured spinal cord were determined by immunofluorescence staining. RESULTS AND CONCLUSION:(1)At 35 days after spinal cord injury,BBB score and inclined plane test score in the riluzole group were higher than those in the model group(P<0.05).(2)At 3 days after spinal cord injury,the protein expressions of NLRP3,cleaved Caspase-1,gasdermin D-N(N-terminal domain),interleukin-1β,and interleukin-18 in the spinal cord homogenate of the riluzole group were significantly lower than those of the model group(P<0.05).(3)At 3 days after spinal cord injury,the fluorescence intensity of NLRP3,Caspase-1,gasdermin D and interleukin-1β in the riluzole group was significantly lower than that in the model group(P<0.05).(4)At day 35 after spinal cord injury,hematoxylin-eosin staining showed that the area of spinal cord injury in the riluzole group was smaller than that in the model group.Electrophysiological tests showed that the latency periods of sensory-evoked potential and motor-evoked potential in the riluzole group were shorter than those in the model group,and the latency period of wave amplitude in the riluzole group was higher than that in the model group.(5)These results suggest that riluzole can promote the repair of injured spinal cord tissue,promote the repair of nerve conduction function,and further promote the recovery of motor function in rats with spinal cord injury,which may be achieved through the regulation of NLRP3 inflammasome and the reduction of microglial pyroptosis.

ObjectiveTo evaluate the clinical efficacy of Chaigui Zengmian prescription （composed of Bupleuri Radix， Codonopsis Radix， Scutellariae Radix， Cinnamomi Ramulus， Paeoniae Radix Alba， Astragali Radix， Atractylodis Macrocephalae Rhizoma， and Scorpio） in treating recurrent respiratory tract infections in children with syndrome of Qi deficiency in lung and spleen and the influences of this prescription on immune factors. MethodSixty-six children suffering from recurrent respiratory tract infections due to Qi deficiency in lung and spleen were selected and randomized into a control group （33 cases） and an observation group （33 cases）. The control group received spleen aminopeptide oral lyophilized powder and the observation group was treated with self-made Chaigui Zengmian prescription granules. Both groups were treated for two months （as two consecutive courses of treatment）. The traditional Chinese medicine （TCM） symptom scores and the serum levels of total immunoglobulins （IgA， IgM， and IgG） and T-helper cells （Th1， Th2， and Th1/Th2） were measured in both groups before and after treatment and then compared. Result① After treatment， the TCM symptom scores of both groups declined and the observation group had lower scores than the control group （P<0.05）. ② The observation group［87.50%（28/32）］ had higher total response rate than the control group［96.88%（31/32）］（Z=7.921，P<0.05）.Also the observation group［28.13%（9/32）］ had more cured children than the control group［6.25（2/32）］ （χ²=5.379，P<0.05） ③ After treatment， the serum levels of IgA， IgM， IgG， Th1， and Th1/Th2 elevated while that of Th2 declined （P<0.05）， and the changes were more obvious in the observation group than in the control group （P<0.05）. ConclusionChaigui Zengmian prescription is effective in treating recurrent respiratory tract infections in children with the syndrome of Qi deficiency in lung and spleen. It can alleviate the symptoms and rectify immune disequilibrium， demonstrating a clinical application value.

BACKGROUND:Neuroinflammation is an important factor leading to secondary spinal cord injury,and microglia/macrophage pyroptosis is a significant part of post-spinal cord injury neuroinflammation.Studies have shown that microglia/macrophage undergoes pyroptosis after spinal cord injury,but the regulatory mechanism of circular RNA(circRNA)in microglia/macrophage pyroptosis after spinal cord injury remains unclear. OBJECTIVE:To investigate the role and mechanism of circRNA0005512 in regulating microglia/macrophage pyroptosis after spinal cord injury. METHODS:Female Wistar rats were divided into sham group and spinal cord injury group.Motor function was evaluated using the Basso,Beattie,and Bresnahan(BBB)scale.Cavity volume was assessed by hematoxylin-eosin staining.Differential expression of circRNA in spinal cord tissue was screened using RNA-sequencing and circ0005512 was validated by real-time PCR.Immunofluorescence,western blot assay,ELISA,and real-time PCR were performed to detect cell pyroptosis in the rats and lipopolysaccharide-induced microglial cell line HAPI cell models.Gene knockdown was used to confirm the regulatory role of circRNA0005512 in microglia/macrophage pyroptosis. RESULTS AND CONCLUSION:(1)Seven days after spinal cord injury,evident cavities were observed at the injury site.Immediately after spinal cord injury,the motor function of rats was completely lost.Over time,the motor function of rats in the spinal cord injury group gradually partially recovered,and there was a significant difference in BBB scores compared to the sham group.(2)Circ0005512 was significantly upregulated according to the results of the RNA-sequencing and confirmed in both the animal and cell models.(3)Immunofluorescence,western blot assay,real-time PCR,and ELISA confirmed the significant upregulation of cell pyroptosis markers(NLRP3,GSDMD,and caspase-1)in spinal cord injury tissue and lipopolysaccharide-induced HAPI cells.(4)In the cell model,knockdown of circ0005512 resulted in significantly decreased levels of cell pyroptosis marker-NLRP3.(5)The results above indicate that circ0005512 promotes pyroptosis in microglia/macrophages after spinal cord injury.

ObjectiveTo observe the effects of Cuscutae Semen on the learning and memory ability， N6-methyladenosine （m⁶A）-related modification enzymes and total m⁶A level in hippocampus of the offspring of fear-damaged pregnant rats. MethodForty-five pregnant rats were randomly divided into blank group， model group and Cuscutae Semen group. From the 1^st day to the 19^th day of pregnancy， rats in the model group and the Cuscutae Semen group were induced by observing electric shock of other rats. The Cuscutae Semen group was treated with 5 g·kg^-1·d^-1 Cuscutae Semen decoction （ig）， while the other two groups were treated with the same amount of purified water. The offspring were assigned following the grouping method of their maternal generation. The behavioral changes of the offspring were tested by Morris water maze on 21^st day after birth， and the development of hippocampal neurons was observed by transmission electron microscopy. The mRNA and protein expression levels of methyltransferase-like 3 （METTL3）， METTL14， Wilms tumor 1 associated protein （WTAP）， fat mass and obesity-associated protein （FTO） and Alk B homologue 5 （ALKBH5） were detected by Real-time polymerase chain reaction （Real-time PCR）， Western blot and immunohistochemistry （IHC）. The total content of m⁶A in hippocampus was determined by high performance liquid chromatography tandem mass spectrometry （LC-MS/MS）. ResultCompared with the conditions in the blank group， the average latency duration in the model group was prolonged， and the number of entries in the target quadrant， the target quadrant duration and the number of crossing the platform were decreased （P<0.01）. Additionally， the model group had seriously damaged structure of hippocampal CA1 and CA3 neurons， swollen mitochondria， expanded endoplasmic reticulum， and small number of synapses with some having blurred structure， and the expression levels of METTL3， METTL14， FTO， ALKBH5 as well as the total m⁶A level were lower than those in the blank group （P<0.05，P<0.01）. Compared with the model group， the Cuscutae Semen group had shortened average latency duration， increased number of entries in the target quadrant， target quadrant duration and number of crossing the platform （P<0.01）， alleviated damage of hippocampal CA1 and CA3 neurons， fine structure of mitochondrial and endoplasmic reticulum， and clear， intact and dense synapses. And the expression levels of METTL3， METTL14， FTO as well as the total level of m⁶A were up regulated， while the expression level of ALKBH5 was down regulated in the Cuscutae Semen group （P<0.05，P<0.01）. ConclusionCuscutae Semen improved the learning and memory ability of the offspring of the rats affected by fear damaging kidney during pregnancy， protected hippocampal neurons， and up-regulated the expression levels of METTL3， METTL14， FTO and the total m⁶A level in hippocampus.

Claudin18.2(CLDN18.2)is a tight junction protein that is overexpressed in a variety of solid tumors such as gastrointestinal cancer and oesophageal cancer.It has been identified as a promising target and a potential biomarker to diagnose tumor,evaluate efficacy,and determine patient prognosis.TST001 is a recombinant humanized CLDN18.2 antibody that selectively binds to the extracellular loop of human Claudin18.2.In this study,we constructed a solid target radionuclide zirconium-89(89Zr)labled-TST001 to detect the expression of in the human stomach cancer BGC823CLDN18.2 cell lines.The[89Zr]Zr-des-ferrioxamine(DFO)-TST001 showed high radiochemical purity(RCP,＞99％)and specific activity(24.15±1.34 GBq/μmol),and was stable in 5％human serum albumin,and phosphate buffer saline(＞85％RCP at 96 h).The EC50 values of TST001 and DFO-TST001 were as high as 0.413±0.055 and 0.361±0.058 nM(P＞0.05),respectively.The radiotracer had a significantly higher average standard uptake values in CLDN18.2-positive tumors than in CLDN18.2-negative tumors(1.11±0.02 vs.0.49±0.03,P=0.0016)2 days post injection(p.i.).BGC823CLDN18.2 mice models showed high tumor/muscle ratios 96 h p.i.with[89Zr]Zr-DFO-TST001 was much higher than those of the other imaging groups.Immunohistochemistry results showed that BGC823CLDN18.2 tumors were highly positive(+++)for CLDN18.2,while those in the BGC823 group did not express CLDN18.2(-).The results of ex vivo biodistribution studies showed that there was a higher distribution in the BGC823CLDN18.2 tumor bearing mice(2.05±0.16％ID/g)than BGC823 mice(0.69±0.02％ID/g)and blocking group(0.72±0.02％ID/g).A dosimetry estimation study showed that the effective dose of[89Zr]Zr-DFO-TST001 was 0.0705 mSv/MBq,which is within the range of acceptable doses for nuclear medicine research.Taken together,these re-sults suggest that Good Manufacturing Practices produced by this immuno-positron emission tomog-raphy probe can detect CLDN18.2-overexpressing tumors.

[摘 要] 目的：探讨鱼藤素通过调控miR-520a-3p表达对卵巢癌SKOV3细胞增殖和凋亡的影响。方法：将SKOV3细胞分为对照组（鱼藤素0 μmol/L）、鱼藤素低剂量（5 μmol/L）、中剂量（10 μmol/L）、高剂量（20 μmol/L）组，miR-NC组、过表达miR-520a-3p组，鱼藤素+anti-miR-NC组、鱼藤素+anti-miR-520a-3p组。CCK-8法、细胞集落形成实验、FCM以及qPCR法分别检测SKOV3细胞的增殖抑制率、细胞克隆形成数、凋亡率以及miR-520a-3p表达水平。结果：与对照组比较，鱼藤素（低、中、高剂量）组SKOV3细胞增殖抑制率、凋亡率、miR-520a-3p表达水平均显著升高（均P<0.05），细胞克隆形成数显著减少（P<0.05）。与miR-NC组比较，过表达miR-520a-3p组SKOV3细胞的增殖抑制率、凋亡率均显著升高（均P<0.05），细胞克隆形成数显著减少（P<0.05）。与鱼藤素+anti-miR-NC组比较，鱼藤素+anti-miR-520a-3p组SKOV3细胞的增殖抑制率、凋亡率均显著降低（均P<0.05），细胞克隆形成数显著增多（P<0.05）。结论：鱼藤素通过增加miR-520a-3p表达抑制卵巢癌SKOV3细胞的增殖能力，并诱导其凋亡。

Objective:To explore the effect of combining extracorporeal shockwave therapy (ESWT) with electromyographic biofeedback on the motor functioning of children with spastic cerebral palsy.Methods:Fifty children with spastic cerebral palsy (2-7 years old) were randomized into a control group and an experimental group, each of 25. Both groups received routine rehabilitation and electric stimulation therapy with electromyographic biofeedback, while the experimental group was additionally provided with ESWT. Before and after 8 weeks of treatment, integral electromyograms were recorded and the root mean square values of the electromyogram recorded from the tibialis anterior muscle were computed. Lower limb spasticity and motor ability were evaluated by using the Modified Ashworth Scale (MAS), the Modified Tardieu Scale (MTS), and the D and E regions of the Gross Motor Function Measurement Scale (GMFM-88).Results:After the treatment the results of both groups had improved significantly in terms of the surface electromyography of the tibialis anterior muscle, MAS scores, R1 in the MTS and the difference between R2 and R1, and the GMFM-88 regional scores. All of the improvements were significantly greater in the experimental group than in the control group, on average.Conclusions:Combining ESWT with EMG biofeedback can better relieve lower limb spasticity and improve the motor functioning of children with spastic cerebral palsy.

Objective To explore the correlation between polarization status of microglia/macrophages(MG/MP)in brain tissue and edema around hematoma in patients with acute cerebral hemorrhage.Methods A total of 52 patients with acute intracerebral hemorrhage admitted to Anyang People's Hospital from December 2020 to November 2022 were selected as the research subjects.All patients underwent craniotomy to remove hematoma,and the normal brain tissue in the cortical area that was not invaded by the hematoma and the fragmented brain tissue around the hematoma(brain tissue around the hematoma)on the surgical pathway were obtained.The expression levels of inflammatory factors such as interleukin(IL)-1β,IL-6,tumor necrosis factor-α(TNF-α),IL-10 and transforming growth factor-β(TGF-β)protein in brain tissue were detected by Western blot.The expression levels of IL-1 β,IL-6,TNF-α,IL-10 and TGF-β mRNA in brain tissue were detected by fluorescence quantitative polymerase chain reaction.The levels of M1-type and M2-type MG/MP in brain tissue was detected by immunofluorescence confocal technique.CT images data of patients before operation were collected and the relative-erihema-tomal edema(r-PHE)was calculated.The patients were divided into high r-PHE group(2.0≤ r-PHE＜2.5)and low r-PHE group(1.5＜r-PHE＜2.0)according to r-PHE.The relative expression of IL-1 β,IL-6,TNF-α,IL-10 and TGF-β mRNA in brain tissue around the hematoma of patients between the high r-PHE group and the low r-PHE group was compared.Results The relative expressions of IL-1 β,IL-6,TNF-α protein and mRNA in brain tissue around the hematoma were significantly higher than those in the normal brain tissues(P＜0.05),but there was no significant difference in the relative expressions of IL-10 and TGF-β protein and mRNA between the brain tissue around the hematoma and the normal brain tissue(P＞0.05).The levels of M1 type and M2 type MG/MP in the brain tissue around the hematoma were significantly higher than those in normal brain tissue(P＜0.05).The relative expressions of IL-1β,IL-6 and TNF-α mRNA in the brain tissue around the hematoma of patients in the high r-PHE group were significantly higher than those in the low r-PHE group(P＜0.05),and there was no significant difference in the relative expressions of TGF-β and IL-10 mRNA in the brain tissue around the hematoma of patients between the two groups(P＞0.05).Conclusion The levels of pro-inflammatory factors and M1-type MG/MP are increased in the brain tissue around the hematoma in patients with acute cerebral hemorrhage,and the degree of polarization of M1-type MG/MP is consistent with the degree of edema around hematoma after cerebral hemorrhage.

Major challenges for cancer treatment are how to effectively eliminate primary tumor and sufficiently induce immunogenic cell death (ICD) to provoke a robust immune response for metastasis control. Here, a self-assembled cascade bioreactor was developed to improve cancer treatment with enhanced tumor penetration and synergistic therapy of starvation, chemodynamic (CDT) and photothermal therapy. Ultrasmall FeS-GOx nanodots were synthesized with glucose oxidase (GOx) as template and induced by paclitaxel (PTX) to form self-assembling FeS-GOx@PTX (FGP)

Objective To investigate the relationship between parotid image texture and acute radiation xerostomia (grade) during radiotherapy in patients with head and neck cancer.The mathematical model was established to predict the severity of radiation dry mouth in the early stage.Methods 23 patients with head and neck cancer treated with radiotherapy were observed.The degree of xerostomia was evaluated according to RTOG criteria.The weekly validated CT images of these patients during radiotherapy were collected and transmitted to the MIM system to outline the parotid gland,and an internal analysis program was developed in MATLAB (R2013a).The changes of texture features of weekly parotid CT images during radiotherapy were analyzed,including mean CT value (MCTN),standard deviation (STD),skewness,kurtosis,entropy and volume.The mathematical model was established,and the KNN method was used to optimize the model and predict the level of xerostomia.Results There was no significant correlation among the changes of MCTN,volume and the degree of xerostomia (P ＞ 0.05).However,according to the weekly changes of MCTN and volume,the model was established to predict the grade of xerostomia with an accuracy of 99％.Conclusions The changes of parotid gland MCTN and volume were significantly correlated with acute radiation xerostomia during radiotherapy for head and neck cancer,and the MCTN changes can be used to predict the severity of xerostomia in the early stage.