Exploring the Dual Subjectivity of Kindergarten's Self-made Play and Teaching Aids Activities from the Perspective of Children's Play Development in China

The joint development of teachers and young children is the basis for promoting the development of high-quality preschool education in the new era,and encouraging early childhood teachers to make playthings according to local conditions is both the fine tradition of China's early childhood education and the direct orientation of China's preschool education policy.Homemade play and teaching aids are some of the methods often used in kindergartens,which skillfully combine young children's daily life objects with toys and early childhood teaching,not only enriching the collective activities of young children but also cultivating children's abilities in various aspects.Kindergarten homemade"play and teaching aids"for early childhood teachers and children have their special educational significance and value,therefore,this paper is based on the children and teachers double subject basis,discussing the homemade"play and teaching aids"for children and teachers of the double subject meanings,and puts forward the realization of the This paper discusses the double-subjective meaning of homemade teaching aids for children and teachers on the basis of the double-subjective foundation of children and teachers,and puts forward specific suggestions for realizing the double-subjectivity of kindergarten homemade teaching aids to enrich the theoretical value and practical significance of homemade teaching aids,to provide theoretical references and support for the relevant staff.

Observation on Curative Effect of Self-made Wubeizi Decoction Combined with rhaFGF in Promoting Postoperative Wound Healing in Patients with Anal Fistula

[Objectives]To explore the curative effect of Self-made Wubeizi Decoction combined with recombinant human acidic fibroblast growth factor(rhaFGF)in promoting postoperative wound healing in patients with anal fistula.[Methods]A total of 82 patients with anal fistula who underwent anal fistula resection+use of setons in Luodian Hospital during January and March of 2020 were randomly divided into observation group and control group with 41 cases in each group.The control group was given rhaFGF external application regimen,and the observation group was treated with Self-made Wubeizi Decoction on the basis of the control group.After 3 weeks of treatment,the differences in curative effects of TCM syndromes were compared between the two groups.Besides,the changes in TCM symptom scores,inflammatory mediators[interleukin-12(IL-12),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)],anorectal functions[anal resting pressure(ARP),maximal systolic pressure(MSP)and maximum tolerated volume(MTV)of the anal canal],quality of life[GQOLI-74 scores]were compared.[Results]After 3 weeks of treatment,the total effective rate of the observation group was significantly higher than that of the control group(P<0.05);the levels of TCM symptom scores,IL-12,TNF-α,IFN-γ,ARP,MSP,and MTV in both groups were significantly lower than those before treatment,and these levels in the observation group was significantly lower than that in the control group at the same time(P<0.05).The GQOLI-74 scores of both groups were significantly higher than those before treatment,and the observation group was significantly higher than the control group(P<0.05).[Conclusions]The Self-made Wubeizi Decoction combined with rhaFGF therapy has a significant effect in promoting postoperative wound healing in patients with anal fistula.It can effectively inhibit the local inflammation of patients,facilitate the recovery of anorectal function and improve the quality of life,thus has high clinical application value.

Process Analysis of Mobile Fermentation of Spent White Hypsizygus marmoreus Substrate with Self-made Mobile Fermentation Bag

The self-made mobile fermentation bag for spent white Hypsizygus marmoreus substrate was developed, and the relevant fermentation process was studied. The results showed that under the condition of single addition of Lactobacillus,nitrogen-free extract was degraded into lactic acid, leading to decreased pH value.On day 10, the fermentation effect reached the best with Lactobacillus abundance of 5.12×10~7/ml, lactic acid content of 0.48% and strong acid flavor. At this time, the fermentation material was moist without mildew and agglomeration, and was suitable for livestock and poultry. However, after 10 days, undesirable acids and mildew generated, a large amount of lactic acid bacteria died, and the fermentation material turned black and agglomerated, and became unsuitable for feeding livestock and poultry. Throughout the fermentation process, the pH value first decreased continuously until to 4.0, and then remained stable. During the fermentation of spent white H. marmoreus substrate, the nitrogen-free extract and crude fiber contents decreased, the crude protein content increased, while other indicators remained unchanged.

Isolation and Identification of Cancer Stem Cells from Human Osteosarcom by Serum-free Three-dimensional Culture Combined with Anticancer Drugs

The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate gel.Single-cell alginate gel was cultured with serum-free DMEM/F12 medium.Epirubicin(0.8μg/mL)was added to the medium to enrich CSCs.After cultured conventionally for 7 to 10 days,most of cells suspended in ...

Serum-free culture of H pylori intensifies cytotoxicity

AIM： To perform a long culture passage of H pylori without serum, taking into account its cytotoxicity and the presence of the probable new cytotoxic factor. METHODS： One sample of H pylon 60190 （ATCC 49503） was grown on Brain Heart Infusion （BHI） agar containing 0.5% 2,6-di-O-methyl-β-cyclodextrin without any serum, being passaged 70-100 times every 3-4 d for approximately 2 h, while another sample of H pylori contained 70 mL/L fetal calf serum without 2,6-di-O- methyl-β-cyclodextrin. Their supernatant and extract after 16 h in culture were evaluated for changes in cell morphology and for cell viability using HeLa cells. Furthermore, the characteristics of the probable cytotoxic factor in the extract were examined on partial purification studies and its oytotoxicity was evaluated in various human cells. RESULTS： The supernatant and the extract of the bacterium grown on serum-free medium had strong cytotoxicity compared with those grown on serumcontaining medium. They irreversibly damaged HeLa cells without vacuolation that was altogether different from that of the bacterium when grown with serum. Their cytotoxicity was easily measured by cell viability assay. The probable cytotoxic factor partially purified and detected by chromatography had characteristics difference from that of vacuolating toxin and a broad cytotoxicity toward various cell lines. CONCLUSION： Serum-free long culture method of H pylorl makes its supernatant and its extract cytotoxic enough to be easily measured by cell viability assay. The probable cytotoxic factor has a unique characteristic and might be a new cytotoxin.

Severe skeletal bimaxillary protrusion treated with micro-implants and a self-made four-curvature torquing auxiliary: A case report

BACKGROUND Bimaxillary protrusion is a clinically common dentofacial deformity,particularly among Chinese patients.This kind of malformation can severely affect facial esthetics and,even in mild cases,is difficult to correct without surgery.Unfortunately,many patients abandon treatment because of fear of surgery.Here,we describe a case of severe skeletal bimaxillary protrusion treated with nonsurgical orthodontic treatments,highlighting an alternative treatment option.CASE SUMMARY A 31-year-old woman wished to address a severe protrusion profile(approximately 8 mm overbite)and gummy smile.Cephalometric analysis and superimposition showed a severe skeletal class II pattern with a mandibular retrusion,and proclined and protrusive mandibular incisors.Panoramic radiograph showed a missing mandibular right third molar.A diagnosis of severe bimaxillary dentoalveolar protrusion was made.Taking into account the patient’s fear of orthognathic surgery,she accepted the proposed alternative treatment using micro-implants and a self-made four-curvature torquing auxiliary.The treatment allowed for maximal en masse anterior tooth retraction,proper relocation of incisors,and alleviation of the skeletal class II pattern.Esthetically,the patient’s lip protrusion was significantly decreased as was the overjet(from 10.5 mm to 1.8 mm),and the results remained stable throughout the 2-year followup.CONCLUSION Nonsurgical treatment using micro-implants and a four-curvature torquing auxiliary may benefit severe cases of skeletal bimaxillary protrusion in adults.

Cholesterol requirement for growth of rodent parental myeloma cells in serum-free medium

Cholesterol,a major lipid component of plasma membrane,is thoughtto have profound effects on the structure and function of cells.Mostanimal tis-sues are capable of synthesizing cholesterol de novo from acetate;however,thereare relatively few mammalian cells in vitro expressing an absolute requirement foran exogenous source of cholesterol.In this paper,it was shown that IR983F(983)rat myeloma cells and P3X63-AgS-U1(P3U1)and P3X63-Ag8.653(653)mousemy eloma cells which had been cultivated in serum-free medium for more than 6months required cholesterol in vitro for growth in serum-free medium.Optimalgrowth of P3U1,653 and 983 occurred in cholesterol concentration of 5,10 and15μg/ml,respectively.Moreover,it was demonstrated that the cholesterol couldbe replaced by human low density lipoprotein in a concentration of 10μg/ml butnot by mevalonic acid lactone.In contrast to the parental myeloma cells,hybridoma cells derived from the mouse myeloma cells which had been cultivatedin serum-free medium for more than 6 months did not require cholesterol.

Some biological characteristics of hybridomas and parental myelomas cultivated in serum-free medium for long-term period

It has been developed in this laboratory that a serum-free medium, designated asDMI,is adaptive for long-term culture,freezing and resurgence in liquid nitrogen,and cloningof hybridoma and parental myelomas as well as for cell fusions.In this report,it is describedthat the myeloma cells grown in DMI for more than 3 months maintained their biological charac-teristics such as induction of aseites and subcutaneous somatic tumor in BALB/c mice and toler-ance toward 8-azaguanine,ete..However,the ability of the tumor cells to attach to glass wallwas decreased.The selecting assay of these cells in HAT medium(medium containing hypoxan-thine,aminopterin and thymidine)showed that the death time in DMI was similar to that in theconventional 15％ newborn calf serum-supplemented medium(RPS15).The hybridoma cellsadapted in DMI secrete monoclonal antibodies in quantities comparable to those produced inRPS15.

INFLUENCE OF MIXING DEVICE ON SERUM-FREE CULTIVATION OF INSECT CELLS IN SPINNER FLASKS

A culture system was developed and successfully employed for the serum-free cultivation of Tn-5B1-4 (Tn5 ) insect cells. With our adaptation procedure, it was possible to obtain cells fully adapted to serum-free media in stationary T-flasks and then enable these adapted cells to grow well in spinner flasks immediately. The spinner "ask with special stirring design proved tO provide favorable culture environment that made it desirable for use in the serum-free cultivation of Tn5 cells even at low seeding density.

Self-made wire loop snare successfully treats gastric persimmon stone under endoscopy

BACKGROUND Large gastric persimmon stones are generally resistant to standard endoscopic treatments.We applied an alternative endoscopic method using a hand-made snare for the treatment of large gastric phytobezoars.AIM To explore the clinical efficacy of a self-made wire loop snare to treat giant gastric persimmon stones.METHODS A retrospective study evaluated the clinical data of 38 patients with gastroliths admitted to Taihe Hospital in Shiyan City,Hubei Province,China,between March 2015 and October 2020.The patients were divided into observation(n=23)and control(n=15)groups.Patients in the observation group were treated with selfmade wire loop snares for lithotripsy,and patients in the control group were treated with traditional foreign body forceps,snares,injection needles,and other tools.Successful stone removal,treatment time,and hospital stay were compared.RESULTS The average operating time was significantly shorter(P<0.001)in the observation group(53.4 min)than that in the control group(172.8 min).The average hospital stay of the observation group(5.4 d)was significantly shorter(P<0.001)than that in the control group(10.3 d).Successful one-time treatment was significantly more frequent(P<0.001)in the observation group(87%)than in the control group(7%).CONCLUSION Self-made guidewire loop snares were successfully used to treat gastrolithiasis, and were significantly more effective than foreign body forceps, snares, and other traditionalmethods.

Novel sericin-based hepatocyte serum-free medium and sericin's effect on hepatocyte transcriptome

AIM To develop a novel hepatocyte serum-free medium based on sericin,and to explore the effect of sericin on the hepatocyte transcriptome.METHODS A controlled trial comparing novel serum-free medium and other media: C3 A cells were cultured in our novel serum-free medium,Hepato ZYME,complete medium(DMEM/F12 with 100 m L/L FBS),and DMEM/F12,andthen cell attachment,proliferation,and function as well as the biocompatibility of the media were assessed.A comparative study of serum-free media with or without 2 mg/m L sericin: the effect of sericin on C3 A growth was assessed by cell viability and proliferation,the effect of sericin on C3 A cell cycle distribution was determined by flow cytometry,and the effect of sericin on the C3 A transcriptome was assessed by gene-chip array and RT-q PCR.RESULTS More C3 A cells attached to the plate containing our serum-free medium than to those containing Hepato ZYME and DMEM/F12 at 24 h post-seeding.Both the viability and proliferation rate of C3 A cells in sericin-based serum-free medium were superior to those of cells in Hepato ZYME and DMEM/F12(P < 0.001).The content of albumin and urea in our serum-free medium was significantly higher than that in Hepato ZYME and DMEM/F12 throughout the whole culture period(P < 0.001) and was similar to that in complete medium at day 3,4,and 5.In part 2,cell viability and proliferation were greater in the presence of 2 mg/m L sericin(P < 0.001),as was the proportion of cells in S phase(16.21% ± 0.98% vs 12.61% ± 0.90%,P < 0.01).Gene-chip array analysis indicated that the expression of CCR6,EGFR,and FOS were up-regulated by 2 mg/m L sericin,and RT-q PCR revealed that the expression of CCR6,EGFR,FOS,AKT1,JNK1,NFk B1,MMP-9,MEK2,ERK1/2 and MYC was upregulated by 2 mg/m L sericin(P < 0.05).CONCLUSION We developed a novel hepatocyte serum-free medium.Sericin probably enhances cell attachment through the CCR6-Akt-JNK-NF-κB pathway and promotes cell proliferation through CCR6-mediated activation of the ERK1/2-MAPK pathway.

Protective Effects of Self-made Compound Taoren Danshen Decoction on Rats with Acute Liver Injury

[Objectives] To study the protective effects of self-made Compound Taoren Danshen Decoction( CTDD) on acute liver injury induced by D-galactosamine in rats,and to explore its mechanism. [Methods]An acute liver injury model was established by intraperitoneal injection of 500 mg/kg of D-galactosamine. The ALT,AST,MDA,SOD and GSH-Px in serum,as well as serum TNF-α,IL-1β and IL-6 levels were measured,and liver tissue lesions were observed under microscope. [Results] CTDD can significantly reduce ALT,AST and MDA levels,increase SOD,GSH-Px activity,and significantly reduce serum TNF-α,IL-1β and IL-6 levels,and improve liver tissue lesions.[Conclusions]CTDD has a protective effect on D-galactosamine-induced acute liver injury in rats,and its mechanism may be related to inhibition of oxidative stress and inflammatory reaction.

CLONAL PROLIFERATION AND LONG-TERM CULTURE OF MALIGNANT LYMPHOMA CELLS UNDER SERUM-FREE CULTURE CONDITIONS

We developed a serum-free culture system that promoted the growth of B cell colonies in peripheral blood, bone marrow, lymph nodes and cerebrospinal fluid (CSF) from 7 out of 8 patients with non-Hodgkin's lymphomas of B cell type. The culture cells were pretreated with or without galactose oxi-dase (GO) prior to plating. Colony growth was best supported with BCGF. A moderate increment was observed with rIL-3, as well as rIL-1β and even to a lesser degree, by rlL-2, while B cell stimulating factor-2 (rBCSF-2) and rlL-1β did not show significant activity. rGM-CSF and rG-CSF had little effect, while rM-CSF enhanced the formation of lymphoma colonies. The cells from different patients had different requirements for Staphylococcus aureus protein A and GO pretreatment. It reflected the differences in activation and differentiation status and surface properties of lymphoma cells from different patients. The cells from CSF of one patient were successfully maintained in serum-free culture medium supplemented with 10% BCGF or 5% PHA-LCM for more than 4 months. The long-term culture cells were EBV negative, phenotypically consistent with B cells and gene rearrangements for JH, Kappa and myc. This serum-free culture system allowed extensive analysis of the growth requirements for clonogenic precursors.

A PRIMARY OBSERVATION OF TPA EFFECT ON SSV-NIH3T3 CELLS IN SERUM-FREE MEDIUM

The effect of TPA, a potent tumor promoter, on SSV-NIH3T3 cells in serum-free medium was investigated. TPA stimulated DNA synthesis of SSV-NIH3T3 cells on the third day of culture in SFM. In SDS-PAGF of medium conditioned by TPA-treated SSV-NIH3T3 cells (in SFM+TPA), the amounts of four proteins of 31.0 Kd, 28.5 Kd, 25.5 Kd and 13.5 Kd strikingly increased over that of non-TPA-treated counterpart (in SFM). The PDGF-like activity was also detected in CM of SFM+TPA. When insulin and EGF were drown off the SFM+TPA (SFM-Ins-EGF+TPA), TPA lost its ability to stimulate DNA synthesis of SSV-NIH3T3 cells on the third day and SDS-PAGE of the conditioned medium showed that the amounts of the four proteins noted above grately reduced. However, cells in SFM-Ins-EGF+TPA were in almost the same growth condition as cells in complete SFM+TPA on the third day of culture. Results were discussed in the paper.

Observation on the Effect of Umbilical Therapy of Herbal Medicine Combined with Self-Made Dysmenorrhea Paste on Primary Dysmenorrhea

Objective:To investigate the clinical effect of self-made dysmenorrhea paste combined with umbilical therapy of herbal medicine on primary dysmenorrhea.Methods:160 patients with primary dysmenorrhea were recruited from Changle People’s Hospital.Participants were randomly divided into control group and treatment group,with 80 cases in each group.The control group were administered the dysmenorrhea paste sold on the market for 5 days during three menstruation cycles,and the treatment group was given the umbilical therapy of herbal medicine combined with self-made dysmenorrhea paste.The outcome was the intensity of menstrual pain measured by using the visual analogue scale,TCM symptom score and dysmenorrhea symptom score of patients,responder rate.The clinical outcomes were measured on each menstrual cycle at baseline,treatment course(3 cycles),and follow-up period.Results:In this study,there was no significant difference in age and severity of disease between the two groups before treatment.After treatment,VAS score,TCM symptom score and dysmenorrhea symptom score of patients in both groups decreased.The total effective rate was 90.75%in the treatment group and 81.25%in the control group.Conclusion:The treatment of primary dysmenorrhea patients with self-made dysmenorrhea paste combined with application of drugs and moxibustion naval-button method has a significant clinical effect.This method can reduce the pain of patients,improve the quality of life of patients,and homemade dysmenorrhea paste has a good taste,long shelf life,easy to carry,and is worthy of promotion and application.

Effect of self-made exhaust plaster on patients with gastrointestinal dysfunction after gynecological operation

Objective:To explore the effect of self-made exhaust patch(SMEP)on gastrointestinal dysfunction after gynecological operation.Methods:A total of 200 patients with gynecological operation in the gynecological ward of Shandong Changle People's Hospital were randomly divided into the control group(n=100)and the treatment group(n=100).The recovery time of bowel sounds,the time of first anal exhaust,the duration of abdominal distention,the duration of abdominal pain,the incidence of abdominal distention and abdominal pain and linical efficacy of SMEP were observed.Results:Compared with the control group,the recovery time of gastrointestinal function and the first anal exhaust time in the treatment group were shorter;the duration of abdominal distention and abdominal pain in the treatment group were shorter than those in the control group(P<0.05);Cure rate of abdominal distention in the treatment is 62.5%and cure rate of abdominal pain in the treatment is 60%within 72 hours after operation,which is higher than that in the control group(44.%,45.9%).Conclusion:The results showed that the self-made exhaust patch can improve the clinical symptoms of patients after gynecological abdominal surgery,and the self-made exhaust patch had a significant effect on gastrointestinal dysfunction after gynecological surgery,which was worthy of clinical promotion.

Clinical observation on TCM fumigation and washing for treatment of diabetic peripheral neuropathy by self-made water-washing footbath

the quality of life seriously,and even lead to more servere complications.On the base of the well-controlled blood glucose,improving the symptoms of both lower limbs timely will has positive significance for enhancing the life quality of patients and preventing complications of limbs.This article makes a comprehensive study on the application of self-made water-washing footbath in the treatment of DPN by Chinese medicine fumigation.From March 2018 to April 2018 in our hospital 20 cases of DPN patients were collected and they were divided into observation group(treated by self-made water-washing footbath with herbs)and control group(treated by conventional barrels),10 cases in each.The results showed that Chinese medicine fumigation on both lower limbs has clinical curative effect to improve the symptoms of arthromyodynia caused by diabetes.It provide an alternative method of external treatment for DPN patients with numbness in both lower limbs which could alleviate their suffering and improve the quality of life significantly.

人胚胎干细胞无血清无饲养层培养体系的建立

目的拟建立人胚胎干细胞无血清无饲养层的培养体系。方法将干细胞分别置于Knockout培养基(第1组)、自制无血清培养基(第2组)和添加bFGF及肝素的改良无血清培养基(第3组)中培养25代后观察比较3组干细胞形态学变化、克隆数目及大小。并借助细胞免疫组化、流式细胞术和体外拟胚体形成等方法检测干细胞的未分化状态和全能性。结果传代25次后,第2组克隆逐渐分化。第1组和第3组的干细胞均呈典型的人胚胎干细胞形态特征;细胞表达Nanog,Oct-4,Tra-1-60,SSEA-4,但不表达SSEA-1;流式检测也显示SSEA-4高表达,SSEA-1低表达;体外分化能形成拟胚体。通过单位视野下克隆个数及大小的比较,结果显示第2组与第1组和第3组间的差异显著。结论本研究通过改良本实验室已获得国家专利的无血清培养基,初步建立了人胚胎干细胞无血清无饲养层培养体系,培养效果与公认培养体系无明显差异。

两种药物局部治疗萎缩性阴道炎108例疗效观察

目的比较自配药Ⅱ号与欧维婷乳膏通过阴道局部给药治疗萎缩性阴道炎的临床效果。方法采用随机对照方法,将108例萎缩性阴道炎患者随机分为两组,Ⅱ号药组54例,欧维婷组54例。分别在阴道局部给予自配药Ⅱ号和欧维婷乳膏进行治疗,比较两组临床治疗效果。结果Ⅱ号药组总有效率为88.88%,欧维婷组为64.82%,两组相比差异有统计学意义(P<0.05);Ⅱ号药组治疗显效时间(6.04±1.55)d,欧维婷组(7.35±1.91)d,两组相比差异有统计学意义(P<0.05)。结论自配药Ⅱ号治疗萎缩性阴道炎的效果优于欧维婷乳膏,且配方简单、价廉,症状缓解快,效果好,患者乐于接受,值得推广。

Serum-free culture of dendritic cells from patients with chronic myeloid leukemia in vitro and estimation of their cytotoxicity

OBJECTIVE: To establish a serum-free culture system of dendritic cells (DCs) from chronic myeloid leukemia (CML) cells so that DCs vaccine may be applied to the adoptive immunotherapy of CML in the near future. METHODS: Fetal calf serum, serum-free medium and autologous serum were used for culture of DCs. The usage of cytokines was classified into two groups: group A (stem cell factor, granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4) and group B (granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4). The phenotypes of DCs were analyzed by using indirect immunofluorescence and flow cytometry. Mixed leukocyte responses were performed by methyl thiazolyl tetrazolium (MTT) assay. Chromosome analysis of DCs can be achieved by displaying G banding. T cells from CML patients were stimulated with autologous DCs and T-cell cytotoxicity was measured by (MTT) assay. RESULTS: CD34(+) cells or mononuclear cells were obtained from peripheral blood or bone marrow samples of eight patients of chronic-phase CML. Group A of serum-free medium was better than group B in expansion of total cell numbers and the rate of DCs. These results of serum-free medium were not significantly different from those of fetal calf serum medium, but the results of autologous serum medium were inferior to two groups above. The expression of major histocompatibility complex class II antigen on the surface of DCs was notable (> 50%), but the expression of CD83 and the costimulatory molecules CD86 was not noticeable (10% - 50%). Although CD1a(+)/CD14(-) DCs were potent stimulators of allogeneic lymphocytes, expansion of T cells from normal volunteers were not significant (average 27.2 fold at DCs: T cells ratio of 1:10). At day 12, CD1a(+) cells from three patients were studied by displaying G banding and Ph(+) cells in these populations were 100%, 98% and 60%, respectively. At an effector: target ratio of 40:1, 32% to 45% cytotoxicity was noted with DC-stimulated T cells against autologous leukemia cells. CONCLUSIONS: A stable serum-free culture system of CML-DCs was established. The expression of CD83 and CD86 on the surface of CML-DCs and DCs' potent stimulation of allogeneic lymphocytes were not notable. DCs in CML patients can be derived from the malignant clone and these malignant DCs could induce anti-leukemic reactivity in autologous T lymphocytes without the necessity for additional exogenous antigens.