Sheep pox is widespread worldwide and is the most severe animal pox virus infection. This study aimed to identify the key microRNAs (miRNAs) differentially expressed in the exosomes of sheep poxvirus-infected cells and their target genes and related pathways and provide a theoretical basis for an in-depth understanding of the molecular mechanisms of sheep poxvirus-infected cells. In this study, the differentially expressed miRNAs were verified by quantitative polymerase chain reaction (qPCR), and the target genes of miRNAs were predicted and analyzed by bioinformatics. The qPCR results showed that the expression trends of oar-miR-21, oar-miR-10b, oar-let-7f, oar-let-7b, and oar-miR-221 were consistent with the sequencing results. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes results showed that differentially expressed miRNAs were mainly involved in the immune system processes of the Arf6 downstream pathway. The target genes Reactome pathways were mainly enriched in the RAC1 GTPase cycle, CDC42 GTPase cycle, RHO GTPase cycle, RHOV GTPase cycle, and post-transcriptional silencing of small RNAs. The transcription factors SP4, NKX6-1, MEF2A, SP1, EGR1, and POU2F1 that may be connected to sheep pox virus (SPPV)-infected cells were discovered by transcription factor annotation screening. In conclusion, this study screened for differentially expressed miRNAs in SPPV-infected cells and performed a series of bioinformatic analyses of their target genes to provide a theoretical basis for the molecular mechanism of sheep pox virus infections of cells. The data can be used as basic information in future studies on the defense mechanisms against poxvirus infections.

Objective:To investigate alterations in functional connectivity network and brain function activity in childhood absence epilepsy (CAE) based on neuromagnetic signals by using multi-frequency magnetoencephalography.Methods:Twenty-five drug-naive children diagnosed with CAE from the Affiliated Brain Hospital of Nanjing Medical University and the Affiliated Children′s Hospital of Nanjing Medical University during October 2022 and March 2024 and 25 healthy controls matched for age and sex from community were recruited in this cross-sectional study. The interictal data, ictal data of CAE and healthy control children were collected using a CTF-275 channel magnetoencephalography system. Corrected amplitude envelope correlation was used to construct functional connectivity network, and network-based statistics were used to compare network differences between groups. Relative power spectral density was used to describe the distribution characteristics of whole-brain spectral power. Nonparametric permutation tests were conducted 1 000 times to compare spectral power differences between groups.Results:In terms of functional connectivity, significant increases in network activity were observed in the low-frequency bands (δ, θ) during interictal periods in children with CAE. A sub-network with significantly increased functional connectivity, including key nodes of the default mode network, was observed in the δ band. Compared with interictal periods, functional connectivity in the δ band decreased during absence seizures in children with CAE, while connectivity in the mid-to-high-frequency bands (α-γ2) increased. In terms of spectral power, children with CAE during interictal periods exhibited widespread magnetic source activation in the δ band, activation in parts of the parietal and occipital lobes in the θ band, and significantly decreased magnetic source intensity in most areas of the parietal, occipital, and temporal lobes in the α-γ2 band. Compared with interictal periods, children with CAE during absence seizures exhibited widespread magnetic source activation in the δ band, and significantly decreased activation in the θ-γ2 band. According to the magnetic source distribution map, during absence seizures, the frontal lobe replaced the parieto-occipital region in cortical activation in the α band.Conclusion:In the analysis of functional network and spectral power based on multi-frequency neuromagnetic signals, the network pattern and magnetic source activation of children with CAE during interictal periods were significantly different from those of healthy children, and there were characteristic changes in neuromagnetic signals during consciousness impairment caused by absence seizures in children with CAE.

Objective To observe the effects of acupuncture at acupoints along the meridians on the expression of frontal lobe associated protein kinase in pchlorophenylalanine(PCPA)induced insomnia rats;To explore the mechanism of acupuncture in improving insomnia.Methods Totally 60 rats were randomly divided into blank group,model group,sham acupuncture group,acupuncture group and Western medicine group,with 12 rats in each group.An insomnia model rat was induced by intraperitoneal injection of PCPA.The acupuncture group received acupuncture at"Baihui","Shenmen"(bilateral),and"Sanyinjiao"(bilateral),while the sham acupuncture group only fixed the needle at the corresponding acupoints but did not penetrate the skin,while the model group was only fixed for 30 minutes,the Western medicine group was given a solution of estazolam by gavage for 7 consecutive days,the blank group was not treated.The pentobarbital sodium reversal experiment was used to detect the sleep latency and sleep time of rats,ELISA was used to detect the content of melatonin(MT)in plasma,RT-PCR was used to detect the mRNA expressions of calmodulin dependent protein kinase(CaMK)Ⅱ,protein kinase C(PKC)and p38 mitogen activated protein kinase(MAPK)in frontal lobe tissue,and Western blot was used to detect the protein expressions of CaMKⅡ,PKC and p38MAPK in frontal lobe tissue.Results Compared with the blank group,the sleep latency of the model group rats were significantly prolonged and the sleep time were significantly shortened(P<0.01),the content of MT in plasma was significantly reduced(P<0.01),and the mRNA and protein expressions of CaMKⅡ and PKC in frontal lobe tissue were significantly decreased(P<0.01),the mRNA and protein expressions of p38MAPK were significantly increased(P<0.01).Compared with the model group,the sleep latency were significantly shortened and the sleep time were significantly prolonged in the acupuncture group and the Western medicine group(P<0.01),the plasma MT content significantly increased(P<0.01),the mRNA and protein expressions of CaMKⅡ and PKC in frontal lobe tissue significantly increased(P<0.05),and the mRNA and protein expression of p38MAPK significantly decreased(P<0.01).Compared with the sham acupuncture group,the sleep latency shortened and sleep time were significantly prolonged in the acupuncture group and the Western medicine group(P<0.01),plasma MT content significantly increased(P<0.01),PKC mRNA expression in frontal lobe tissue significantly increased(P<0.01),and p38MAPK mRNA expression significantly decreased(P<0.01).There was no statistically difference in various indicators between the acupuncture group and the Western medicine group(P>0.05).Conclusion Acupuncture at acupoints along the meridians can shorten the sleep latency and prolong sleep time,improve sleep status by regulating the expression of associated protein kinase in frontal lobe of insomnia rats.

Sheep pox is widespread worldwide and is the most severe animal pox virus infection. This study aimed to identify the key microRNAs (miRNAs) differentially expressed in the exosomes of sheep poxvirus-infected cells and their target genes and related pathways and provide a theoretical basis for an in-depth understanding of the molecular mechanisms of sheep poxvirus-infected cells. In this study, the differentially expressed miRNAs were verified by quantitative polymerase chain reaction (qPCR), and the target genes of miRNAs were predicted and analyzed by bioinformatics. The qPCR results showed that the expression trends of oar-miR-21, oar-miR-10b, oar-let-7f, oar-let-7b, and oar-miR-221 were consistent with the sequencing results. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes results showed that differentially expressed miRNAs were mainly involved in the immune system processes of the Arf6 downstream pathway. The target genes Reactome pathways were mainly enriched in the RAC1 GTPase cycle, CDC42 GTPase cycle, RHO GTPase cycle, RHOV GTPase cycle, and post-transcriptional silencing of small RNAs. The transcription factors SP4, NKX6-1, MEF2A, SP1, EGR1, and POU2F1 that may be connected to sheep pox virus (SPPV)-infected cells were discovered by transcription factor annotation screening. In conclusion, this study screened for differentially expressed miRNAs in SPPV-infected cells and performed a series of bioinformatic analyses of their target genes to provide a theoretical basis for the molecular mechanism of sheep pox virus infections of cells. The data can be used as basic information in future studies on the defense mechanisms against poxvirus infections.

Sheep pox is widespread worldwide and is the most severe animal pox virus infection. This study aimed to identify the key microRNAs (miRNAs) differentially expressed in the exosomes of sheep poxvirus-infected cells and their target genes and related pathways and provide a theoretical basis for an in-depth understanding of the molecular mechanisms of sheep poxvirus-infected cells. In this study, the differentially expressed miRNAs were verified by quantitative polymerase chain reaction (qPCR), and the target genes of miRNAs were predicted and analyzed by bioinformatics. The qPCR results showed that the expression trends of oar-miR-21, oar-miR-10b, oar-let-7f, oar-let-7b, and oar-miR-221 were consistent with the sequencing results. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes results showed that differentially expressed miRNAs were mainly involved in the immune system processes of the Arf6 downstream pathway. The target genes Reactome pathways were mainly enriched in the RAC1 GTPase cycle, CDC42 GTPase cycle, RHO GTPase cycle, RHOV GTPase cycle, and post-transcriptional silencing of small RNAs. The transcription factors SP4, NKX6-1, MEF2A, SP1, EGR1, and POU2F1 that may be connected to sheep pox virus (SPPV)-infected cells were discovered by transcription factor annotation screening. In conclusion, this study screened for differentially expressed miRNAs in SPPV-infected cells and performed a series of bioinformatic analyses of their target genes to provide a theoretical basis for the molecular mechanism of sheep pox virus infections of cells. The data can be used as basic information in future studies on the defense mechanisms against poxvirus infections.

Sheep pox is widespread worldwide and is the most severe animal pox virus infection. This study aimed to identify the key microRNAs (miRNAs) differentially expressed in the exosomes of sheep poxvirus-infected cells and their target genes and related pathways and provide a theoretical basis for an in-depth understanding of the molecular mechanisms of sheep poxvirus-infected cells. In this study, the differentially expressed miRNAs were verified by quantitative polymerase chain reaction (qPCR), and the target genes of miRNAs were predicted and analyzed by bioinformatics. The qPCR results showed that the expression trends of oar-miR-21, oar-miR-10b, oar-let-7f, oar-let-7b, and oar-miR-221 were consistent with the sequencing results. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes results showed that differentially expressed miRNAs were mainly involved in the immune system processes of the Arf6 downstream pathway. The target genes Reactome pathways were mainly enriched in the RAC1 GTPase cycle, CDC42 GTPase cycle, RHO GTPase cycle, RHOV GTPase cycle, and post-transcriptional silencing of small RNAs. The transcription factors SP4, NKX6-1, MEF2A, SP1, EGR1, and POU2F1 that may be connected to sheep pox virus (SPPV)-infected cells were discovered by transcription factor annotation screening. In conclusion, this study screened for differentially expressed miRNAs in SPPV-infected cells and performed a series of bioinformatic analyses of their target genes to provide a theoretical basis for the molecular mechanism of sheep pox virus infections of cells. The data can be used as basic information in future studies on the defense mechanisms against poxvirus infections.

Sheep pox is widespread worldwide and is the most severe animal pox virus infection. This study aimed to identify the key microRNAs (miRNAs) differentially expressed in the exosomes of sheep poxvirus-infected cells and their target genes and related pathways and provide a theoretical basis for an in-depth understanding of the molecular mechanisms of sheep poxvirus-infected cells. In this study, the differentially expressed miRNAs were verified by quantitative polymerase chain reaction (qPCR), and the target genes of miRNAs were predicted and analyzed by bioinformatics. The qPCR results showed that the expression trends of oar-miR-21, oar-miR-10b, oar-let-7f, oar-let-7b, and oar-miR-221 were consistent with the sequencing results. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes results showed that differentially expressed miRNAs were mainly involved in the immune system processes of the Arf6 downstream pathway. The target genes Reactome pathways were mainly enriched in the RAC1 GTPase cycle, CDC42 GTPase cycle, RHO GTPase cycle, RHOV GTPase cycle, and post-transcriptional silencing of small RNAs. The transcription factors SP4, NKX6-1, MEF2A, SP1, EGR1, and POU2F1 that may be connected to sheep pox virus (SPPV)-infected cells were discovered by transcription factor annotation screening. In conclusion, this study screened for differentially expressed miRNAs in SPPV-infected cells and performed a series of bioinformatic analyses of their target genes to provide a theoretical basis for the molecular mechanism of sheep pox virus infections of cells. The data can be used as basic information in future studies on the defense mechanisms against poxvirus infections.

OBJECTIVE To investigate the role of clinical pharmacists in the treatment of a patient with Epstein-Barr （EB） virus encephalitis. METHODS Clinical pharmacist participated in drug diagnosis and therapy for a patient with EB virus encephalitis. According to the physiological characteristics of the disease and the pharmacokinetic-pharmacodynamic characteristics of antibiotics， clinical pharmacists suggested that the dose should be adjusted as ceftriaxone 2 g， q12 h+meropenem 2 g， q8 h. Based on the uncontrolled infection of the patient， pharmacists suggested that ceftriaxone should be stopped and vancomycin 1 million U and q12 h should be used as alternative therapy. According to the results of etiology， pharmacists suggested that acyclovir should be discontinued and replaced with ganciclovir 5 mg/kg， q12 h. The electrolyte disturbance of the patient may be adverse drug reactions caused by Mannitol injection， it was recommended to stop the drug. RESULTS The clinician followed the advice of the clinical pharmacists. After treatment， the patient improved and was discharged. CONCLUSIONS Clinical pharmacists can carry out pharmaceutical care for patients with EB virus encephalitis， assist physicians in optimizing the treatment plan of patients， and ensure the effectiveness and safety of drug treatment.

AIM: To compare the clinical efficacy of conbercept and aflibercept in the treatment of wet age-related macular degeneration(wARMD)based on 4 consecutive intravitreal injections.METHODS: The clinical data of 108 patients(108 eyes)who were diagnosed as wARMD and treated with intravitreal injection at our hospital from January 2019 to January 2021 were retrospectively analyzed. They were divided into conbercept group(54 cases, 54 eyes)and aflibercept group(54 cases, 54 eyes)according to the different injectable drugs. All patients received intravitreal injection once a month, with four consecutive injections. Follow up for 12mo to observe best corrected visual acuity(BCVA), central macular thickness(CMT), complications and recurrence before and after injection.RESULTS: BCVA and CMT of patients in the two groups at 1, 2, 5 and 8mo after injection had no between-group differences(P>0.05), but both were significantly improved compared with those before injection(P<0.05). By the end of follow-up, conjunctival hemorrhage occurred in 2 eyes of the conbercept group at the early stage, and increased intraocular pressure and conjunctival hemorrhage occurred respectively in 2 eyes of the aflibercept group. There were no serious complications related to drug injection such as retinal detachment, complicated cataract, endophthalmitis and retinal pigment epithelial tear in the two groups, and there was no difference in the recurrence rate between the two groups(7% vs. 6%, P=1.000).CONCLUSION: On the basis of continuous 4 times of intravitreal injection, both conbercept and aflibercept are safe and effective in the treatment of wARMD, and the efficacy is even.

Objective: To explore the effects of three-dimensional (3D) bioprinting gelatin methacrylamide (GelMA) hydrogel loaded with nano silver on full-thickness skin defect wounds in rats. Methods: The experimental research method was adopted. The morphology, particle diameter, and distribution of silver nanoparticles in nano silver solution with different mass concentrations and the pore structure of silver-containing GelMA hydrogel with different final mass fractions of GelMA were observed by scanning electron microscope and the pore size was calculated. On treatment day 1, 3, 7, and 14, the concentration of nano silver released from the hydrogel containing GelMA with final mass fraction of 15% and nano silver with final mass concentration of 10 mg/L was detected by mass spectrometer. At 24 h of culture, the diameters of inhibition zone of GelMA hydrogel containing final mass concentration of 0 (no nano silver), 25, 50, and 100 mg/L nano silver against Staphylococcus aureus and Escherichia coli were detected. Fibroblasts (Fbs) and adipose stem cells (ASCs) were isolated respectively by enzymatic digestion using the discarded prepuce after circumcision from a 5-year-old healthy boy who was treated in the Department of Urology of the Second Affiliated Hospital of Zhejiang University School of Medicine in July 2020, and the discarded fat tissue after liposuction from a 23-year-old healthy woman who was treated in the Department of Plastic Surgery of the Hospital in July 2020. The Fbs were divided into blank control group (culture medium only), 2 mg/L nano sliver group, 5 mg/L nano sliver group, 10 mg/L nano sliver group, 25 mg/L nano sliver group, and 50 mg/L nano sliver group, which were added with the corresponding final mass concentrations of nano sliver solution, respectively. At 48 h of culture, the Fb proliferation viability was detected by cell counting kit 8 method. The Fbs were divided into 0 mg/L silver-containing GelMA hydrogel group, 10 mg/L silver-containing GelMA hydrogel group, 50 mg/L silver-containing GelMA hydrogel group, and 100 mg/L silver-containing GelMA hydrogel group and then were correspondingly treated. On culture day 1, 3, and 7, the Fb proliferation viability was detected as before. The ASCs were mixed into GelMA hydrogel and divided into 3D bioprinting group and non-printing group. On culture day 1, 3, and 7, the ASC proliferation viability was detected as before and cell growth was observed by live/dead cell fluorescence staining. The sample numbers in the above experiments were all 3. Four full-thickness skin defect wounds were produced on the back of 18 male Sprague-Dawley rats aged 4 to 6 weeks. The wounds were divided into hydrogel alone group, hydrogel/nano sliver group, hydrogel scaffold/nano sliver group, and hydrogel scaffold/nano sliver/ASC group, and transplanted with the corresponding scaffolds, respectively. On post injury day (PID) 4, 7, 14, and 21, the wound healing was observed and the wound healing rate was calculated (n=6). On PID 7 and 14, histopathological changes of wounds were observed by hematoxylin eosin staining (n=6). On PID 21, collagen deposition of wounds was observed by Masson staining (n=3). Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, Bonferroni correction, and independent sample t test. Results: The sliver nano particles in nano silver solution with different mass concentrations were all round, in scattered distribution and uniform in size. The silver-containing GelMA hydrogels with different final mass fractions of GelMA all showed pore structures of different sizes and interconnections. The pore size of silver-containing GelMA hydrogel with 10% final mass fraction was significantly larger than that of silver-containing GelMA hydrogels with 15% and 20% final mass fractions (with P values both below 0.05). On treatment day 1, 3, and 7, the concentration of nano silver released from silver-containing GelMA hydrogel in vitro showed a relatively flat trend. On treatment day 14, the concentration of released nano silver in vitro increased rapidly. At 24 h of culture, the diameters of inhibition zone of GelMA hydrogel containing 0, 25, 50, and 100 mg/L nano silver against Staphylococcus aureus and Escherichia coli were 0, 0, 0.7, and 2.1 mm and 0, 1.4, 3.2, and 3.3 mm, respectively. At 48 h of culture, the proliferation activity of Fbs in 2 mg/L nano silver group and 5 mg/L nano silver group was both significantly higher than that in blank control group (P<0.05), and the proliferation activity of Fbs in 10 mg/L nano silver group, 25 mg/L nano silver group, and 50 mg/L nano silver group was all significantly lower than that in blank control group (P<0.05). Compared with the that of Fbs in 0 mg/L silver-containing GelMA hydrogel group, the proliferation activity of Fbs in 50 mg/L silver-containing GelMA hydrogel group and 100 mg/L silver-containing GelMA hydrogel group was all significantly decreased on culture day 1 (P<0.05); the proliferation activity of Fbs in 50 mg/L silver-containing GelMA hydrogel group was significantly increased (P<0.05), while the proliferation activity of Fbs in 100 mg/L silver-containing GelMA hydrogel group was significantly decreased on culture day 3 (P<0.05); the proliferation activity of Fbs in 100 mg/L silver-containing GelMA hydrogel group was significantly decreased on culture day 7 (P<0.05). The proliferation activity of ASCs in 3D bioprinting group show no statistically significant differences to that in non-printing group on culture day 1 (P>0.05). The proliferation activity of ASCs in 3D bioprinting group was significantly higher than that in non-printing group on culture day 3 and 7 (with t values of 21.50 and 12.95, respectively, P<0.05). On culture day 1, the number of dead ASCs in 3D bioprinting group was slightly more than that in non-printing group. On culture day 3 and 5, the majority of ASCs in 3D bioprinting group and non-printing group were living cells. On PID 4, the wounds of rats in hydrogel alone group and hydrogel/nano sliver group had more exudation, and the wounds of rats in hydrogel scaffold/nano sliver group and hydrogel scaffold/nano sliver/ASC group were dry without obvious signs of infection. On PID 7, there was still a small amount of exudation on the wounds of rats in hydrogel alone group and hydrogel/nano sliver group, while the wounds of rats in hydrogel scaffold/nano sliver group and hydrogel scaffold/nano sliver/ASC group were dry and scabbed. On PID 14, the hydrogels on the wound surface of rats in the four groups all fell off. On PID 21, a small area of wounds remained unhealed in hydrogel alone group. On PID 4 and 7, the wound healing rates of rats in hydrogel scaffold/nano sliver/ASC group were significantly higher than those of the other three groups (P<0.05). On PID 14, the wound healing rate of rats in hydrogel scaffold/nano sliver/ASC group was significantly higher than the wound healing rates in hydrogel alone group and hydrogel/nano sliver group (all P<0.05). On PID 21, the wound healing rate of rats in hydrogel alone group was significantly lower than that in hydrogel scaffold/nano sliver/ASC group (P<0.05). On PID 7, the hydrogels on the wound surface of rats in the four groups remained in place; on PID 14, the hydrogel in hydrogel alone group was separated from the wounds of rats, while some hydrogels still existed in the new tissue of the wounds of rats in the other three groups. On PID 21, the collagen arrangement in the wounds of rats in hydrogel alone group was out of order, while the collagen arrangement in the wounds of rats in hydrogel/nano sliver group, and hydrogel scaffold/nano sliver/ASC group was relatively orderly. Conclusions: Silver-containing GelMA hydrogel has good biocompatibility and antibacterial properties. Its three-dimensional bioprinted double-layer structure can better integrate with new formed tissue in the full-thickness skin defect wounds in rats and promote wound healing.
Male ; Rats ; Animals ; Humans ; Hydrogels/pharmacology* ; Bioprinting ; Metal Nanoparticles ; Rats, Sprague-Dawley ; Silver/pharmacology* ; Soft Tissue Injuries ; Anti-Bacterial Agents