BACKGROUND:The regulation of M1/M2 polarization direction of macrophages is particularly critical in tissue engineering applications,and timely regulation can minimize proinflammatory,anti-inflammatory,or tissue healing responses. OBJECTIVE:To implant lentivirus-mediated miRNA-378a macrophage strain complex collagen to detect the expression level of immune regulation in the in vivo environment,and further clarify the influence of miRNA-378a in promoting macrophage M2 polarization in immune regulation and tissue repair in the in vivo environment. METHODS:Lentivirus-mediated miRNA-378a overexpressing macrophage cell lines and negative control virus macrophage lines were amplified and screened,and the macrophage lines were recovered and cultured together with collagen sponge to form a composite scaffold,which was divided into the following groups:(1)Positive group:miRNA-378a overexpressing macrophage-collagen sponge composite;(2)negative group:negative control of virus-mediated miRNA-378a macrophage-collagen sponge composite;(3)control group:macrophage-collagen sponge;(4)blank control group:collagen sponge.The cell density,phenotype,and adhesion of each group were observed by immunofluorescence and scanning electron microscopy.The cells were implanted into the subcutaneous model of the back of mice,and the mice were sacrificed 4 and 7 days after modeling.The direction of macrophage polarization in the collagen sponge composite of macrophages with miRNA-378a overexpression mediated by lentivirus and its effect on immune regulation and tissue repair were analyzed by gross observation,hematoxylin-eosin staining,MASSON staining,and immunohistochemistry. RESULTS AND CONCLUSION:(1)Under immunofluorescence microscopy,the macrophage cell lines in each group were observed to form a composite scaffold with the collagen sponge.(2)Under scanning electron microscope,lentivirus-mediated miRNA-378a macrophages in the positive group proliferated in cell density,had spherical,elliptic and polygonal differentiation,and had more pseudofeet than other groups.(3)Under general observation,the overall 7-day healing was better than that at 4 days.Lentivirus-mediated miRNA-378a macrophages in the positive group healed better than other groups regardless of 4 and 7 days.(4)Lentivirus-mediated miRNA-378a macrophages in the positive group under hematoxylin-eosin staining and MASSON staining had more amounts of fibrocytes,capillaries,fibroblasts,and collagen fiber hyperplasia.(5)Immunohistochemistry showed that lentivirus-mediated miRNA-378a macrophages in the positive group were more positive in 4-and 7-day M2 polarized cells than in other groups.The macrophages of the control and negative groups in 4-and 7-day M2 polarized cells were greater than that of the blank control group.There was no statistical difference between the control group and the negative group.The number of stained cells in the positive,negative,and control groups regardless of 4 and 7 days was higher than that in the blank control group,and the positive group>negative group ≈ control group>blank control group.(6)It is concluded that macrophages with miRNA-378a overexpression have a large amount of fibroblasts,capillaries,fibroblasts,and collagen fiber hyperplasia in vivo,which has a positive effect on tissue repair,and can promote the polarization of macrophages towards M2 type and inhibit the polarization of M1 type,thus contributing to reducing the inflammatory response of the body.

BACKGROUND:M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing.Therefore,how to regulate M2 polarization of macrophages has been a hot research topic in recent years,and some miRNAs have been found to have this function. OBJECTIVE:To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line. METHODS:The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ.Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages.By transfection with lentivirus as the vector of overexpression of miR-378a,the stable expression of miR-378a cell lines was screened.Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ.Macrophage M2 polarization was induced by interleukin-4.The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay.qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines. RESULTS AND CONCLUSION:(1)The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization.(2)Compared with the non-transfected group,the expressions of proinflammatory cytokine-induced nitric oxide synthase,tumor necrosis factor-α,interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group(P<0.05);the levels of inducible nitric oxide synthase,tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased(P<0.05).(3)Compared with the non-transfected group,the expressions of CD206,interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased(P<0.05);the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased(P<0.05)in the miR-378a transfection group.(4)It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages.

Objective To explore the effect of imitation of bioelectricity stimulus combined with estrogen and progesterone on promoting endometrial repair and preventing intrauterine adhesion after missed abortion.Methods A total of 120 patients with missed abortion treated in Shenzhen Nanshan Maternal and Child Health Care Hospital from June 2021 to December 2022 were selected as the research objects.The patients were numbered according to the order of admission,and they were divided into control group (n=60) and combined group (n=60) according to the random number table method.Both groups were treated with curettage after medical abortion.The control group was treated with estrogen and progesterone,and combined group was treated with imitation of bioelectricity stimulus on the basis of control group.The duration of postoperative abdominal pain,visual analogue scale (VAS) score of abdominal pain 72 hours after operation,endometrial thickness,the end-systolic peak/end-diastolic peak (S/D),pulsation index (PI) and resistance index (RI) on the day after operation,15 days after operation and 15 days after the first menstrual onset were compared between two groups.The incidence of intrauterine adhesions in two groups was counted.Results The duration of postoperative abdominal pain and VAS score of abdominal pain at 72 hours after operation in combined group were significantly lower than those in control group (P<0.05).The endometrial thickness of combined group was higher than that of control group 15 days after operation and 15 days after the first menstrual onset,and S/D,PI and RI were lower than those of control group (P<0.05).During the treatment,the incidence of intrauterine adhesion in combined group was 3.33％,which was significantly lower than 13.33％ in control group (P<0.05).Conclusion Compared with the use of estrogen and progestins alone,imitation of bioelectricity stimulus combined with estrogen and progesterone can more effectively repair the endometrium and prevent the occurrence of intrauterine adhesion in patients with missed abortion,which is worthy of clinical promotion.

Objective: To examine the impact of physical intelligence teaching on the function of children s sensory integration, so as to provide reference for promoting the development of sensory integration system. Methods: From February to May 2023, the intervention was implemented for 12 weeks among 136 children aged 4-5 (68 in the intervention group and 68 in the control group). The intervention group received situational and game based physical intelligence teaching, the control group received sports game teaching according to the original curriculum objectives of the kindergarten. Intervention was administered 3 times a week for 40 minutes each time. The sensory integration ability of the intervention group and the control group were evaluated before and after the intervention with Chi square test and t test. Results: The vestibular sensation, proprioception and tactile sensation of between boys and girls in the intervention group were significantly improved compared with before intervention (boys：44.14±11.52 vs. 53.34± 9.49 ，44.57±12.76 vs. 50.54±11.86，49.31±12.18 vs. 55.00±10.24,girls：46.00±11.01 vs. 54.58±10.06，48.79±13.17 vs. 53.64±11.97，52.67±11.67 vs. 56.91±10.42, t =-3.24，-2.49，-2.09，-5.24，-12.94，-2.56， P <0.05). There was no significant difference in vestibular sensation between boys and girls in the control group (boys：45.91±11.66 vs. 46.31± 11.20，girls：48.27±13.56 vs. 48.45 ±13.54, t =-0.87，-0.07， P >0.05), but there was a significant improvement in proprioception and tactile sensation in both boys and girls (boys：46.63±11.76 vs. 48.06±11.69，51.63±11.98 vs. 52.40±12.18，girls：50.45±12.16 vs. 51.67± 12.03 ，53.36±12.48 vs. 54.39±12.57， t =-3.36，-2.08，-4.66，-2.86， P <0.05). After the intervention, compared with the control group, the vestibular sensation of both boys and girls significantly improved ( t=2.83, 2.08, P <0.05), with exception of proprioception and tactile sensation ( t =0.88,0.67,0.97,0.88, P >0.05). In the experimental group, the number of normal boys increased from 12 to 24, while the number of dysfunctional boys decreased from 23 to 11, with a statistically significant difference ( χ 2=11.53, P <0.01). There was no statistically significant difference in sensory integration in boys of the control group before and after the experiment ( χ 2= 1.10 , P >0.05). After intervention,the number of normal girls in the experimental group increased from 15 to 27, while the number of dysfunctional girls decreased from 18 to 6, with a statistically significant difference ( χ 2=10.39, P < 0.05 ). There was no statistically significant difference in sensory integration in girls from the control group before and after the experiment ( χ 2=2.08, P > 0.05 ). Conclusion Physical intelligence teaching can effectively improve children s sensory integration ability, especially for vestibular function.

Early diagnosis and treatment is the key to improve the prognosis and reduce the disability of Wilson′s disease. Conventional magnetic resonance imaging sequences are difficult to detect early brain lesions in Wilson′s disease, especially in preclinical patients. Quantitative susceptibility mapping (QSM) is a recently developed magnetic resonance post-processing technique, which is very sensitive to magnetic metal deposition in the brain. QSM can be used for quantitative analysis of metal deposition in the basal ganglia of Wilson′s disease and is expected to be an imaging technique for early diagnosis and condition assessment of Wilson′s disease.

Identifying data-driven biotypes of major depressive disorder (MDD) has promise for the clarification of diagnostic heterogeneity. However, few studies have focused on white-matter abnormalities for MDD subtyping. This study included 116 patients with MDD and 118 demographically-matched healthy controls assessed by diffusion tensor imaging and neurocognitive evaluation. Hierarchical clustering was applied to the major fiber tracts, in conjunction with tract-based spatial statistics, to reveal white-matter alterations associated with MDD. Clinical and neurocognitive differences were compared between identified subgroups and healthy controls. With fractional anisotropy extracted from 20 fiber tracts, cluster analysis revealed 3 subgroups based on the patterns of abnormalities. Patients in each subgroup versus healthy controls showed a stepwise pattern of white-matter alterations as follows: subgroup 1 (25.9% of patient sample), widespread white-matter disruption; subgroup 2 (43.1% of patient sample), intermediate and more localized abnormalities in aspects of the corpus callosum and left cingulate; and subgroup 3 (31.0% of patient sample), possible mild alterations, but no statistically significant tract disruption after controlling for family-wise error. The neurocognitive impairment in each subgroup accompanied the white-matter alterations: subgroup 1, deficits in sustained attention and delayed memory; subgroup 2, dysfunction in delayed memory; and subgroup 3, no significant deficits. Three subtypes of white-matter abnormality exist in individuals with major depression, those having widespread abnormalities suffering more neurocognitive impairments, which may provide evidence for parsing the heterogeneity of the disorder and help optimize type-specific treatment approaches.

OBJECTIVE: To assess the potential of transient expression of recombinant human plasminogen activator (rhPA) in plants as a cost-effective approach for recombinant rhPA production. METHODS: Tobacco mosaic virus-based expression vector pTMV rhPA-NSK and plant binary expression vector pJ Zera-rhPA were constructed by sequence synthesis and subcloning. The two vectors were inoculated on either or leaves agroinfiltration. The expression of recombinant rhPA in leaves was examined using Western blotting and ELISA, and the fibrinolysis activity of plant-produced rhPA was assessed by fibrin agarose plate assay (FAPA). RESULTS: Five to nine days after infiltration with an inoculum containing pTMV rhPA-NSK, necrosis appeared in the infiltrated area on the leaves of both plants, but intact recombinant rhPA was still present in the necrotic leaf tissues. The accumulation level of recombinant rhPA in infiltrated leaves was significantly higher than that in leaves ( < 0.05). The yield of recombinant rhPA was up to 0.6% of the total soluble protein (or about 60.0 μg per gram) in the fresh leaf biomass at 7 days post-inoculation. The plant-derived rhPA was bioactive to convert inactive plasminogen to active plasmin. No necrosis occurred in pJ Zera-rhPA-infiltrated leaves. The Zera-rhPA protein was partially cleaved between the site of Zera tag and rhPA sequence in both leaves. We speculated that the formation of Zera tags-induced particles in the plant cells was a dynamic process of progressive aggregation in which some of the soluble polypeptides were encapsulated in these particles. CONCLUSIONS Enzymatically active recombinant rhPA can be rapidly expressed in tobacco plants using the plant viral ampliconbased system, which offers a promising alternative for cost-effective production of recombinant rhPA.
Humans ; Plant Leaves ; Plants, Genetically Modified ; Plasminogen ; Plasminogen Activators ; metabolism ; Recombinant Proteins ; Tobacco

Neurocognitive deficits are frequently observed in patients with schizophrenia and major depressive disorder (MDD). The relations between cognitive features may be represented by neurocognitive graphs based on cognitive features, modeled as Gaussian Markov random fields. However, it is unclear whether it is possible to differentiate between phenotypic patterns associated with the differential diagnosis of schizophrenia and depression using this neurocognitive graph approach. In this study, we enrolled 215 first-episode patients with schizophrenia (FES), 125 with MDD, and 237 demographically-matched healthy controls (HCs). The cognitive performance of all participants was evaluated using a battery of neurocognitive tests. The graphical LASSO model was trained with a one-vs-one scenario to learn the conditional independent structure of neurocognitive features of each group. Participants in the holdout dataset were classified into different groups with the highest likelihood. A partial correlation matrix was transformed from the graphical model to further explore the neurocognitive graph for each group. The classification approach identified the diagnostic class for individuals with an average accuracy of 73.41% for FES vs HC, 67.07% for MDD vs HC, and 59.48% for FES vs MDD. Both of the neurocognitive graphs for FES and MDD had more connections and higher node centrality than those for HC. The neurocognitive graph for FES was less sparse and had more connections than that for MDD. Thus, neurocognitive graphs based on cognitive features are promising for describing endophenotypes that may discriminate schizophrenia from depression.
Adult ; Algorithms ; Depressive Disorder, Major ; classification ; diagnosis ; Endophenotypes ; analysis ; Female ; Humans ; Machine Learning ; Male ; Markov Chains ; Neuropsychological Tests ; Schizophrenia ; classification ; diagnosis ; Young Adult

Objective To understand the integrated ability of parasitic disease prevention and control of professional person-nel of Jiangsu Province through the contest. Methods Totally 56 players from the whole province were selected,and all the players participated in the contest. The theory knowledge and skill scores were collected and the statistical analyses were con-ducted. Results The average theoretical score of the participants was 88.86±15.56 and the passing rate was 91.1％. The aver-age skill operating score was 69.16 ± 16.01 and the passing rate was 67.9％. The average Plasmodium microscopy score was 16.54±8.09 and the passing rate was 50％. The average helminth egg microscopy score was 34.27±10.66 and the passing rate was 67.9％. There were statistical differences among the age groups and different levels of schistosomiasis endemic situation (F =5.10,6.39,both P<0.01). The theoretical knowledge including schistosomiasis,malaria,hydatid disease and others and the score rates were 91.07％,90.94％,85.83％and 90.93％,respectively. The hydatid disease score rate was lower(χ2=19.17, P<0.01). The radar chart displayed that the score rates of tabletting and microscopy test in Kato-Katz film production ,malaria blood film production and microscopy test were all low. Conclusion In Jiangsu Province,the participants have higher score in the theory test. However,they have lower skill test score,especially in the parasite species identification. The operational skills still need to be strengthened for center for disease control(CDC)participants.

Objective To know the contamination status of Giardia lamblia and Cryptosporidium in drinking water of Jiangsu Province,so as to provide the evidence for producing hygiene and safety drinking water. Methods A total of 28 water plants of 13 cities in Jiangsu Province were selected,and the source water(10 L),chlorinated water(100 L)and tap water(100 L) were collected separately in each site. The water samples were then treated by filtration,washing,centrifuging concentration, immune magnetic separation,and immunofluorescent assay,to detect the numbers of Giardia cysts and Cryptosporidium oocysts. Results Totally 84 samples from 13 cities were collected,including 28 source water,28 chlorinated water and 28 tap water samples. Among the chlorinated water and tap water samples,no Giardia cysts and Cryptosporidium oocysts were found. Howev-er,Giardia cysts were detected in 3(10.71％,3/28)source water samples(Yancheng,Lianyungang,Changzhou cities),with the density of 1 cyst/10 L of all. Cryptosporidium oocysts were also detected in 3(10.71％,3/28)source water samples(Nan-jing,Zhenjiang,Yangzhou cities),with the density of 1 oocyst/10 L of all. Conclusions The source water in partial areas of Ji-angsu Province has been contaminated by Giardia and Cryptosporidium. To ensure the safety of drinking,the regulation of source water and surveillance of drinking water should be strengthened.