BACKGROUND:The micro/nanostructured gradient biomimetic surface of implant materials can simulate the structure of the extracellular environment in human bone tissue,thereby achieving perfect bone integration function.However,further research is needed on the mechanisms by which the surface microstructure of bone implant materials regulates cell function and promotes osteogenesis. OBJECTIVE:To analyze the effect of titanium sheet microstructure surface on osteogenic differentiation of MC3T3-E1 osteoblasts. METHODS:(1)At a constant voltage of 5 V or 20 V,nanotube arrays of different diameters were prepared on the surface of titanium sheets by acid etching and anodic oxidation techniques,and were recorded as group R5 and group R20,respectively.The surface morphology,roughness,and hydrophilicity of pure titanium sheet(without acid etching or anodizing treatment)were measured in group R5 and group R20.(2)MC3T3-E1 osteoblasts of logarithmic growth stage were inoculated on the surface of pure titanium sheets,R5 group and R20 group respectively.After 24 hours of osteogenic induction culture,the expression of mechanical sensitive channel protein 1 was analyzed by RT-PCR and immunofluorescence staining.Osteoblast inducible base with or without the mechanosensitive channel protein 1 activator Yada1 was added,and alkaline phosphatase staining was performed after 7 days of culture.Alizarin red staining was performed after 14 days of culture. RESULTS AND CONCLUSION:(1)The surface of pure titanium sheets was smooth under scanning electron microscope.Relatively uniform and orderly nanotube arrays with average diameters of about 30 nm and 100 nm were observed on the surface of titanium sheets of groups R5 and R20,respectively.The results of scanning electron microscope were further verified by atomic force microscopy.The surface roughness of titanium sheet of group R5 was higher than that of pure titanium(P<0.05),and the water contact angle was lower than that of pure titanium(P<0.05).The surface roughness of titanium sheet in group R20 was higher than that in group R5(P<0.05),and the water contact angle was lower than that in group R5(P<0.05).(2)RT-PCR and immunofluorescence staining showed that the expression of mechanosensitive channel protein 1 in group R5 was higher than that in pure titanium group(P<0.05),and the expression of mechanosensitive channel protein 1 in group R20 was higher than that in group R5(P<0.05).Under the osteogenic induction,compared with the condition without Yada1,there were no significant changes in the activity of alkaline phosphatase and the deposition of calcified nodules in pure titanium group after Yada1 addition,while the activity of alkaline phosphatase and the deposition of calcified nodules in groups R5 and R20 after Yada1 addition were significantly increased(P<0.05).With or without Yada1,the alkaline phosphatase activity and calcified nodule deposition in group R5 were higher than those in pure titanium group(P<0.05),and the alkaline phosphatase activity and calcified nodule deposition in group R20 were higher than those in group R5(P<0.05).(3)The results show that the surface microstructure of titanium sheet can promote the osteogenic differentiation of osteoblast MC3T3-E1 by activating mechanosensitive channel protein 1.

Objective To explore the etiology and pathogenesis of dry eye by studying the distribution pattern of gender,age and traditional Chinese medicine(TCM)syndrome type in dry eye patients and by analyzing their correlation.Methods A total of 244 patients with dry eye who met the inclusion criteria were selected.The distribution of gender,age and TCM syndrome types was statistically analyzed,and then the correlation of TCM syndrome types with gender and age of dry eye patients was explored.Results(1)Of the 244 dry eye patients,96(39.34%)were male and 148(60.66%)were female,the incidence of the female being higher than that of the male.There were 124(50.82%)patients younger than 45 years old,81(33.20%)patients aged 45-60 years old,and 39(15.98%)patients older than 60 years old.The proportion of the patients younger than 45 years old was higher than that of other age groups.(2)Among the 244 patients with dry eyes,89 cases(36.47%)were differentiated as liver and kidney deficiency syndrome,75 cases(30.74%)were differentiated as qi stagnation and blood stasis syndrome,69 cases(28.28%)were differentiated as spleen and kidney deficiency,and 11 cases(4.51%)were differentiated as yin deficiency and damp-heat syndrome.And the occurrence frequency of the above four syndrome types was in descending order.(3)In the dry eye patients of various age groups,patients aged＜45 years old predominantly suffered from qistagnation and blood stasis syndrome,accounting for 41.94%(52/124);patients aged 45-60 years old and those aged＞60 years old predominantly suffered from liver and kidney deficiency syndrome,accounting for 46.91%(38/81)and 53.85%(21/39),respectively.The distribution of TCM syndrome types varied in the patients with different age groups,and the difference was statistically significant(χ2 = 22.128,P＜0.01).(4)In male dry eye patients,qi stagnation and blood stasis syndrome was predominant,accounting for 39.58%(38/96);among female dry eye patients,liver and kidney deficiency syndrome and spleen and kidney deficiency syndrome were prevalent,accounting for 41.89%(62/148)and 31.08%(46/148),respectively.The distribution of TCM syndrome types varied in the patients with different genders,and the difference was statistically significant(χ2 = 82.610,P＜0.01).Conclusion The TCM syndromes of patients with dry eyes are frequently differentiated as liver and kidney deficiency syndrome,followed by the qi stagnation and blood stasis syndrome.The prevalence of dry eyes is related to the gender and age,and gender and age are correlated with the TCM syndrome types to certain extent.

AIM:To explore the protective effect of Xiaoxuming decoction(XXMD)on synaptic plasticity in the context of cerebral ischemia-reperfusion injury following ischemic stroke.METHODS:An oxygen-glucose depriva-tion/reoxygenation(OGD/R)model was employed in vitro using mouse hippocampal neurons(HT22 cells)to simulate ischemia-reperfusion injury.Cell viability was assessed using a CCK-8 assay to determine the optimal XXMD concentra-tion.The HT22 cells were divided into two groups:control and model(OGD/R).Cellular morphological changes were ob-served using an inverted microscope.The levels of IL-1β,IL-6 and TNF-α in the supernatant were quantified by ELISA.Ultrastructural changes were examined by transmission electron microscopy.Immunofluorescence staining was used to de-tect neuron markers NeuN and synaptic proteins NF200 and MAP2.The protein levels of NF200 and MAP2 were analyzed by Western blot.RESULTS:The highest cell survival rate occurred at an XXMD concentration of 100 mg/L(P<0.05).Compared with control group,the cells in model group exhibited round shape and shrinkage,mitochondrial swelling or vacuolization,and a marked decrease in survival rate.There were significant increases in IL-1β,IL-6 and TNF-α levels(P<0.05).Immunofluorescence intensity and protein levels of NeuN,NF200 and MAP2 were notably reduced(P<0.05).Treatment with XXMD improved cell morphology,ultrastructure and survival rate(P<0.05),and decreased in-flammatory factor levels(P<0.05).Compared with model group,the cells in OGD/R+XXMD group showed significantly increased immunofluorescence intensity and protein levels of NeuN,NF200 and MAP2(P<0.05).CONCLUSION:Xiaoxuming decoction may mitigate OGD/R-induced injury,potentially by inhibiting inflammatory responses and enhanc-ing synaptic plasticity.

Berberine is a natural isoquinoline alkaloid that was initially used as a broad-spectrum antibacterial agent in clinical treatment of enteritis,peptic ulcers,chronic gastritis,pneumonia,and other diseases.In recent years,in-depth study of the pharmacological effects of berberine has provided increasing evidence that berberine has neuroprotective effects on ischemic stroke.In this review,we introduce the effect of berberine on risk factors of ischemic stroke and discuss the neuroprotective effects of berberine on various mechanisms of ischemic stroke in detail to provide a reference for clinical and basic research in this field.

The ethics committee of organ transplantation technology and clinical application in a hospital has encountered some difficulties and typical cases in its review work and practice for many years. Sometimes, it is difficult to make a decision in these dilemmas. Based on the previous experience of the hospital in the ethical review of organ donation and transplantation, combined with two typical cases, this paper discussed and analyzed two review points of whether the voluntary unpaid donation and the principle of informed consent were met, and whether the risk-benefit ratio was reasonable, and put forward relevant ethical and legal countermeasure for further research by institutional ethics committees and other parties, in order to provide reference for discussing the practical problems and ethical confusion of ethical review of organ donation and transplantation.

OBJECTIVE To evaluate the efficacy and safety of dimethyl fumarate （DMF） in the treatment of multiple sclerosis （MS）. METHODS Retrieved from CBM， Web of Science， PubMed， the Cochrane Library， Embase， CNKI， Wanfang Data， and VIP， randomized controlled trials （RCTs） about DMF （trial group） versus other drugs or placebo （control group） were collected. After data screening and extraction， quality evaluation， meta-analysis was conducted by using RevMan 5.3 software. RESULTS A total of 6 literature were included， involving 638 patients. Results of meta-analysis showed that the proportion of patients with lesion changes after treatment in the trial group was lower than control group [MD＝－0.65， 95%CI（－1.27， －0.02）， P＝0.04]； there was no statistical significance in recurrence rate [RR＝1.06， 95%CI（0.52，2.17）， P＝0.88]， the proportion of patients with new lesions after treatment [RR＝1.05， 95%CI（0.62，1.80）， P＝0.85]， expanded disability status scale after treatment [MD＝0.02，95%CI （－0.18， 0.23）， P＝0.82]， the incidence of adverse events [RR＝1.33， 95%CI（0.97， 1.84）， P＝0.08] or severe adverse events [RR＝0.95，95%CI（0.48，1.90），P＝0.89] between 2 groups. Results of sensitivity analysis showed the study obtained unstable recurrence rate and the incidence of adverse events， while other results were robust. CONCLUSIONS DMF can control the lesion progression in MS patients to some extent and doesn’t increase the incidence of adverse events and serious adverse events， but there is no significant advantage in reducing the recurrence rate and controlling the disability progression.

Objective To explore the medication rules of Professor HUANG Li for the treatment of recurrent angina pectoris after percutaneous coronary intervention(PCI)for coronary heart disease by data mining method.Methods The prescriptions for effective cases of recurrent angina pectoris after PCI for coronary heart disease treated by Professor HUANG Li in the outpatient department of China-Japan Friendship Hospital were collected.SPSS Statistics 26.0 software and SPSS Modeler 18.0 software were used for frequency statistics,analysis of the therapeutic actions,properties,flavors and meridian tropism of the prescribed herbs as well as association rule analysis,cluster analysis and factor analysis of the herbs.Results A total of 344 Chinese medicine prescriptions were obtained,involving 209 herbs,with a cumulative frequency of 5 874 times.The top 30 Chinese medicinals were named as the high-frequency Chinese medicines,and the herbs with the frequency over 100 times in descending order were Astragali Radix,Chuanxiong Rhizoma,Puerariae Lobatae Radix,Rhodiolae Crenulatae Radix et Rhizoma,Notoginseng Radix et Rhizoma,Poria,Dalbergiae Odoriferae Lignum,Atractylodis Macrocephalae Rhizoma,Curcumae Rhizoma,Sparganii Rhizoma,Dioscoreae Rhizoma,Citri Reticulatae Pericarpium,Pinelliae Rhizoma Praeparatum,Codonopsis Radix,and Glycyrrhizae Radix et Rhizoma.The high-frequency Chinese medicinals were mostly classified as blood-activating and stasis-resolving drugs and qi-replenishing drugs.The medicinal properties of the drugs were characterized by being warm,mild,or cold,the flavors were predominated by being sweet,pungent or bitter,and the medicinals usually had the meridian tropism of the spleen,lung and liver meridians.A total of 30 association rules were mined out,cluster analysis yielded 5 herbal groups,and factor analysis yielded 11 groups of common factors.Conclusion For the treatment of cardiovascular diseases,Professor HUANG Li follows the theory of qi,blood and water,and especially pays more attention to the ascending and descending of qi movement.For qi deficiency and blood stasis contribute to the basic pathogenesis of recurrent angina pectoris after PCI,the therapy of benefiting qi,activating blood and removing stasis is recommended.Moreover,the simultaneous regulation of five zang-organs and simultaneous use of the cold and warm herbs are performed,and the herbs of benefiting qi and invigorating spleen,resolving phlegm and inducing diuresis,tranquilizing mind,promoting qi and dissipating masses,and activating blood to eliminate stasis are used for adjuvant therapy.

Aim To investigate the dynamic changes of neuronal cells at different time points following acute cerebral ischemia-reperfusion injury by establishing a model of brain ischemia-reperfusion injury.Methods Thirty male Sprague-Dawley(SD)rats were ran-domly divided into six groups:sham group and cere-bral ischemia-reperfusion injury(IR)groups at differ-ent time points.Focal cerebral ischemia-reperfusion injury model was established using the middle cerebral artery occlusion(MCAO)technique.The Longa sco-ring method was used to assess neurobehavioral scores in rats.After successful model preparation,routine paraffin sections were made,and TUNEL staining and immunohistochemistry staining with NeuN antibody were performed to observe cell apoptosis and neuronal cell survival,respectively.Immunohistochemistry stai-ning was also performed to investigate the changes in glial fibrillary acidic protein(GFAP)as a marker for astrocytes,ionized calcium-binding adapter molecule 1(IBA-1)as a marker for microglia,and CD31 as a marker for endothelial cells at different time points.Results No significant changes were observed in neu-ronal cells of the sham group at different time points.In the cerebral ischemia-reperfusion injury groups,cell apoptosis was activated at IR3h and increased in quan-tity with morphological damage as time progressed.Ne-uN+neurons showed signs of ischemic injury after IR3h,with abnormal cell morphology.From 12 h,Ne-uN+neurons decreased in a time-dependent manner and reached their peak severity at 24 h.GFAP+astro-cytes decreased significantly after IR3h,while poorly labeled GFAP+astrocytes increased at IR 6 h and al-most disappeared in the infarcted area at 24 h and 48 h.The number of IBA-1+microglia-positive cells de-creased at IR3h,and their volume increased at IR6h.Microglial cell death was observed in the infarcted area at IR12h.CD31+endothelial cells around the infarc-ted cortex and striatum increased significantly after IR3h and persisted until 48 h.Conclusions After cerebral ischemia-reperfusion injury,the number of ap-optotic cells increases with the prolongation of time,and NeuN+neurons exhibit the most severe damage at 24 h.GFAP+astrocytes and microglial cells gradually die over time.The number of CD31+endothelial cells increases significantly around the infarcted cortex and striatum after 3 h of reperfusion and persists until 48 h.

Aim To study the resistance of SIRT3 ex-pression in dopaminergic neurons against the inflamma-tory damage caused by microglia activation and its re-lated mechanism.Methods Dopaminergic neurons(MN9D cells)and microglia(BV-2 cells)were co-cultured to establish an inflammatory injury model in vitro.MN9D cells were divided into the control group,model group,SIRT3 group and SIRT3+PJ34 group.mRNA levels were analyzed by real-time quantitative polymerase chain reaction,cell apoptosis rate was de-tected by flow cytometry,changes in mitochondrial membrane potential were tested by JC-1 method,and the opening of mitochondrial permeability transport pore(mPTP)was analyzed by co-incubation of calce-in-AM and CoCl2.The protein expression was detected by Western blot.Results Compared to the model group,overexpression of SIRT3 in the SIRT3 group significantly reduced the apoptosis rate of MN9D cells.It also led to a significant increase in the expression of SIRT3 and SOD2 genes,as well as a notable decrease in PARP-1,tumor necrosis factor-α,and interleukin 1β(IL-1β)protein expressions.Moreover,it resulted in a substantial reduction in the p-NF-κB p65/NF-κB p65 ratio.There was an improvement observed in mito-chondrial membrane potential along with decreased mPTP opening and ROS production in the SIRT3 group.These differences among these groups were sta-tistically significant(all P＜0.05).After inhibiting PARP-1 activity of MN9D cells in the SIRT3+PJ34 group,except for the insignificant changes in SIRT3 and IL-1 β protein expression,the changing trend of other indicators was further enhanced on the basis of SIRT3 group.The differences between two groups re-mained statistically significant(all P＜0.05).Con-clusions SIRT3 expression can attenuate the inflam-matory damage of dopaminergic neurons induced by microglia activation,and the mechanism may be relat-ed to improving mitochondrial function,inhibiting PARP-1 activity and NF-κB signaling pathway caused by the reduction of ROS production.

Objective To observe the ultrasonic manifestations and possible etiology of fetal tricuspid regurgitation(TR).Methods Totally 717 fetuses diagnosed with TR by prenatal ultrasound were retrospectively enrolled,and the prenatal ultrasonic findings were observed.Based on postpartum follow-up data,the fetuses were divided into physiological TR group(n=468)and pathological TR group(n=249),and those in the latter were further divided into right heart preload increase subgroup(n=76),right heart afterload increase subgroup(n=127)and tricuspid valve structure abnormality subgroup(n=46)according to the possible etiology,and the ultrasonic manifestations were comparatively analyzed.Results In physiological TR group,mild and moderate TR was found in 441(441/468,94.23％)and 27 fetuses(27/468,5.77％),respectively,while no severe TR was noticed.In pathological TR group,significant difference of TR degrees was found among 3 subgroups(x2=37.244,P＜0.001).Mild TR was more common in right heart preload increase subgroup,while moderate and severe TR were more common in the other 2 subgroups.In right heart preload increase subgroup,fetuses with persistent left superior vena cava were more likely to develop mild TR,while those with intact interventricular septum and pulmonary artery occlusion were more likely to develop severe TR in right heart afterload increase subgroup(both P＜0.05).No significant difference of TR degree was found among fetuses with different possible etiology in right heart preload increase subgroup nor right heart afterload increase subgroup(both P＞0.05).In tricuspid valve structure abnormality subgroup,significant differences of TR degrees were found among fetuses with different possible etiologies(P＜0.05),and fetuses with underdeveloped tricuspid valve were more prone to severe reflux(P＜0.05).There were significant differences of reflux velocity of moderate and severe TR among 3 subgroups(F=6.945,P=0.002).Conclusion Fetal TR was mostly physiological.The possible etiologies of pathological TR were variable,including pulmonary valve stenosis,persistent left superior vena cava and tricuspid valve hypoplasia,with different prenatal ultrasonic manifestations.