BACKGROUND:At present,hydroxyapatite has been used more and more widely in the field of facial fillers due to its good biocompatibility and low immunity.The main sources are divided into natural extraction and artificial synthesis.However,there are few comparative studies on natural extraction and artificial synthesis of hydroxyapatite,especially the difference in stimulating collagen secretion between the two.OBJECTIVE:To investigate the physicochemical differences between naturally derived and commercially available synthetic calcium hydroxyapatite particles and promoting collagen secretion.METHODS:(1)Natural hydroxyapatite particles from pig bones and two kinds of commercially available synthetic hydroxyapatite particles(denoted as SYN1 and SYN2)were used to characterize the microstructure and surface element content of the three kinds of materials.(2)The three kinds of materials with different mass concentrations(1,5,and 10 mg/mL)were co-cultured with human skin fibroblasts.Cell proliferation was detected by CCK-8 assay.The three kinds of materials at 5 mg/mL were co-cultured with human skin fibroblasts.Cell adhesion was observed by CCK-8 assay and scanning electron microscopy.The expression of type Ⅰ collagen was detected by RT-PCR,ELISA,and western blot assay.(3)Twelve New Zealand rabbits were selected,of which six were subcutaneously injected with a mixture of natural hydroxyapatite and 2％sodium carboxymethylcellulose gel on the back.The remaining six rabbits were subcutaneously injected with a mixture of SYN2 and 2％sodium carboxymethylcellulose gel on the back.The samples were collected 1 and 3 months after injection and stained with Masson and Sirius red.RESULTS AND CONCLUSION:(1)Scanning electron microscope showed that the crystal grains of natural hydroxyapatite particles were more regular and uniform,with holes evenly distributed between particles.The SYN1 grains were smaller and densely arranged,while the SYN2 grains were irregular.The median particle sizes of natural hydroxyapatite particles,SYN1,and SYN2 were 38,24,and 40 pm,respectively.Natural hydroxyapatite contained Mg and Zn,SYN1 did not contain Mg and Zn,and SYN2 did not contain Zn.(2)CCK-8 assay showed that 1 and 5 mg/mL of natural hydroxyapatite and SYN2 promoted the proliferation of human skin fibroblasts,while 5 and 10 mg/mL of SYN1 inhibited the proliferation of human skin fibroblasts.The number of cells adhered to the surface of natural hydroxyapatite particles was more than that of SYN1 and SYN2,and the cells on the surface of natural hydroxyapatite particles spread well,with visible filopodia.RT-PCR,ELISA,and western blot assay results showed that the expression of type Ⅰ collagen in the natural hydroxyapatite particle group was higher than that in the SYN1 group and the SYN2 group.(3)The results of Masson and Sirius red staining showed that the amount of type Ⅰ collagen in the subcutaneous tissue of the natural hydroxyapatite particle group was greater than that of the SYN2 group 1 and 3 months after injection.(4)The results show that compared with synthetic hydroxyapatite particles,natural hydroxyapatite particles have a richer distribution of trace elements and can better promote fibroblast adhesion,proliferation and collagen regeneration.

Objective To explore the repair effect and mechanism of human amniotic mesenchymal stem cells(hAMSCs)on endometrial injury.Methods hAMSCs were isolated using a two-enzyme digestion and then cultured.The third-passage(P3)cells were harvested to detect the surface markers by flow cytometry and to identify their trilineage differentiation potentials.Eighteen nulliparous female SD rats(8～9 weeks old,weighing 250～280 g)were randomly divided into 3 groups(n=6):normal control group,model group,and hAMSCs group.A rat model of intrauterine adhesions(IUA)was established in SD rats by using curettage combined with lipopolysaccharide(LPS)infection.In 2 weeks after modeling,the hAMSCs group received a bilateral uterine horn transplantation of 0.2 mL hAMSCs(1.0×10? cells/mL),while the model group received a same volume of PBS into both uterine horns.All rats were sacrificed in 2 weeks after transplantation.HE and Masson staining was used to observe endometrial thickness and gland number as well as endometrial fibrosis area.RT-qPCR and Western blotting were performed to detect the mRNA and protein levels of TGF-β1,Smad3,Smad7,epithelial-mesenchymal transition(EMT)markers(E-cadherin,Vimentin),fibrosis factor α-SMA,and endometrial estrogen receptor(ER)and progesterone receptor(PR)in endometrial tissues.Results The obtained cells were identified as hAMSCs due to the characteristics of surface markers and differentiation potentials.Compared with the normal control group,the model group showed decreased endometrial thickness,reduced gland number,increased fibrosis area,and enhanced mRNA and protein levels of fibrosis-related factors TGF-β1,Smad3,Vimentin,and α-SMA(P<0.01),while down-regulation of fibrosis-inhibiting molecule Smad7,the EMT marker E-cadherin,and endometrial receptors ER and PR at both mRNA and protein levels(P<0.01).hAMSCs transplantation increased endometrial thickness and gland number,decreased fibrosis area,and down-regulated mRNA expression of the aforementioned fibrosis-related factors(P<0.01),and up-regulated the mRNA expression levels of Smad7,E-cadherin,ER,and PR(P<0.01).The hAMSCs group also exhibited obviously down-regulated protein levels of TGF-β1,Smad3,and α-SMA(P<0.05),while enhanced protein levels of Smad7 and PR(P<0.05).Conclusion Intrauterine transplantation of hAMSCs can promote the repair of endometrial injury,and inhibits endometrial EMT and fibrosis through the TGF-β1/Smad7 signaling pathway.

BACKGROUND:At present,hydroxyapatite has been used more and more widely in the field of facial fillers due to its good biocompatibility and low immunity.The main sources are divided into natural extraction and artificial synthesis.However,there are few comparative studies on natural extraction and artificial synthesis of hydroxyapatite,especially the difference in stimulating collagen secretion between the two.OBJECTIVE:To investigate the physicochemical differences between naturally derived and commercially available synthetic calcium hydroxyapatite particles and promoting collagen secretion.METHODS:(1)Natural hydroxyapatite particles from pig bones and two kinds of commercially available synthetic hydroxyapatite particles(denoted as SYN1 and SYN2)were used to characterize the microstructure and surface element content of the three kinds of materials.(2)The three kinds of materials with different mass concentrations(1,5,and 10 mg/mL)were co-cultured with human skin fibroblasts.Cell proliferation was detected by CCK-8 assay.The three kinds of materials at 5 mg/mL were co-cultured with human skin fibroblasts.Cell adhesion was observed by CCK-8 assay and scanning electron microscopy.The expression of type Ⅰ collagen was detected by RT-PCR,ELISA,and western blot assay.(3)Twelve New Zealand rabbits were selected,of which six were subcutaneously injected with a mixture of natural hydroxyapatite and 2％sodium carboxymethylcellulose gel on the back.The remaining six rabbits were subcutaneously injected with a mixture of SYN2 and 2％sodium carboxymethylcellulose gel on the back.The samples were collected 1 and 3 months after injection and stained with Masson and Sirius red.RESULTS AND CONCLUSION:(1)Scanning electron microscope showed that the crystal grains of natural hydroxyapatite particles were more regular and uniform,with holes evenly distributed between particles.The SYN1 grains were smaller and densely arranged,while the SYN2 grains were irregular.The median particle sizes of natural hydroxyapatite particles,SYN1,and SYN2 were 38,24,and 40 pm,respectively.Natural hydroxyapatite contained Mg and Zn,SYN1 did not contain Mg and Zn,and SYN2 did not contain Zn.(2)CCK-8 assay showed that 1 and 5 mg/mL of natural hydroxyapatite and SYN2 promoted the proliferation of human skin fibroblasts,while 5 and 10 mg/mL of SYN1 inhibited the proliferation of human skin fibroblasts.The number of cells adhered to the surface of natural hydroxyapatite particles was more than that of SYN1 and SYN2,and the cells on the surface of natural hydroxyapatite particles spread well,with visible filopodia.RT-PCR,ELISA,and western blot assay results showed that the expression of type Ⅰ collagen in the natural hydroxyapatite particle group was higher than that in the SYN1 group and the SYN2 group.(3)The results of Masson and Sirius red staining showed that the amount of type Ⅰ collagen in the subcutaneous tissue of the natural hydroxyapatite particle group was greater than that of the SYN2 group 1 and 3 months after injection.(4)The results show that compared with synthetic hydroxyapatite particles,natural hydroxyapatite particles have a richer distribution of trace elements and can better promote fibroblast adhesion,proliferation and collagen regeneration.

OBJECTIVE To study the effects of Zigui yichong formula on premature ovarian insufficiency （POI） in mice through glycolysis metabolic pathway. METHODS Eighty SPF C57BL/6N female mice were divided into normal group， model group， Zigui yichong formula group （14.175 g/kg）， Zigui yichong formula+2-deoxy-D-arabino-hexose （2-DG） group （Zigui yichong formula 14.175 g/kg + glycolysis inhibitor 2-DG 100 mg/kg）， with 20 mice in each group. Except for the normal group， POI model mice were induced by intraperitoneal administration of cyclophosphamide in the other groups. After the model was successfully established， each group was given corresponding drugs. HE staining was employed to observe the pathomorphological changes in ovarian tissue and to count follicles at all developmental stages； radioimmunoassay was conducted to measure the serum levels of estradiol （E2）， anti-Müllerian hormone （AMH）， and follicle-stimulating hormone （FSH）； TUNEL assay was employed to detect apoptosis in ovarian granulosa cells of mice； the activities of hexokinase （HK）， pyruvate kinase （PK） and lactate dehydrogenase （LDH） were detected by colorimetry； Western blot and real-time fluorescence quantitative PCR were employed to analyze the protein and mRNA expressions of B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， caspase-3， HK2， pyruvate kinase M2 （PKM2）， and lactate dehydrogenase A （LDHA）. RESULTS Compared with model group， the number of primordial follicles， growing follicles， antral follicles and granulosa cells were increased significantly（P＜0.05）， and granulosa cells arranged neatly， but the number of atretic follicles and granulosa cells apoptosis were decreased significantly in Zigui yichong formula group （P＜0.05）； the serum levels of E2 and AMH， the activities of HK， PK and LDH， protein and mRNA expressions of Bcl-2， HK2， PKM2 and LDHA were increased significantly （P＜0.05）； the serum levels of FSH， the protein and mRNA expressions of Bax and caspase-3， Bax/Bcl-2 ratio were decreased significantly （P＜0.05）. 2-DG could reverse the improvement effects of Zigui yichong formula on the above indexes of POI model mice. CONCLUSIONS Zigui yichong formula may inhibit the apoptosis of ovarian granulosa cells， reduce follicle atresia and improve ovarian reserve function by promoting glycolysis levels in POI model mice.

Chinese Journalof Stomatology has issued eighty papers on Chinese stomatology history in seventy years. According to three stages of the journal,statistics of the quantity and themes of the issued papers are compiled: the number of issued papers increased while the theme shifted from Chinese stomatology history before 1912 to Chinese contemporary stomatology history. Research methods shifted from the comparatively scanty summary or induction on literature and cultural relics materials to multidisciplinary approaches. Early researches testified the achievments of senior scholars such as Zhou Dacheng and Mao Xiejun in the field while research teams represented by colleges and research institutions have published varied research topics in recent years. As the research deepens, platforms for issuing stomatology papers have been expanded and more subjects have been cross-disciplined. Varieties of academic monographs on stomatology history and stomatology education history have been published, among which Zhou Dacheng's Historic textual research of Chinese stomatology can be regarded as a milestone in the field. Comparing with other fields of stomatology, current research on stomatology history still has much more gaps to fill. The academic community should put more emphasis on talent training, discipline construction, research approaches, museum construction, as well as the cooperation between Chinese and western medical sciences so that to assist in the long-term development of the research.

Objective This study aims to investigate possible hub genes,associated pathways,and transcription fac-tors between chronic periodontitis(CP)and Parkinson's disease(PD).Methods Gene expression profiles of CP(GSE16134,GSE23586,and GSE10334)and PD(GSE20141 and GSE49036)were downloaded from the gene expres-sion omnibus(GEO)database for differential expression analysis and functional clustering analysis.The protein-protein interaction(PPI)network was constructed,and hub genes were screened by four topological analysis algorithms and modular segmentation.Functional clustering analysis was performed.The hub genes were validated by external datasets of CP and PD,and causal relation was further assessed by Mendelian randomization(MR).Results After merging the data,1 211 differentially expressed genes(DEGs)were screened in the CP datasets;of which,551 were upregulated and 660 were downregulated.A total of 2 407 DEGs were screened in the PD dataset,of which,1 438 were upregulated and 969 were downregulated.The PPI network included 145 nodes and 126 edges.Four hub genes(FCGR3B,PRF1,IL18,and CD33)and three transcription factors(HSF1,HSF2,and HSF4)were finally screened.The relevant pathway was pre-dominantly natural killer(NK)cell-mediated toxic effects.The MR results suggest a possible positive causal relationship between CP and the risk of developing PD.Conclusion This study indicated the probably shared pathophysiology and possible causal relationship between CP and PD and may offer novel concepts and therapeutic targets for future mecha-nistic investigations.

Objective:To investigate the expression of macrophage migration inhibitory factor (MIF) in bone marrow fluid and peripheral blood of patients with acute myeloid leukemia (AML) and its effect on the expression of interleukin-8 (IL-8) in bone marrow mesenchymal stem cells (BM-MSC).Methods:Fifty bone marrow fluid samples and 50 peripheral blood samples were collected from 50 patients with AML diagnosed in the First People's Hospital of Yunnan Province from October 2017 to January 2019, of which 17 patients were newly diagnosed, 26 patients were complete remission (CR), and 7 patients were partial remission (PR) or non-remission (NR). Fifty plasma samples from 50 healthy subjects and 50 bone marrow fluid samples from 50 patients with iron deficiency anemia were used as the controls. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of MIF protein in the samples, and the relationship between MIF expression level and clinicopathological characteristics of AML patients was analyzed. BM-MSC was successfully isolated and cultured from 42 bone marrow fluid samples of AML patients, the suitable samples for experiment were chosen and divided into BM-MSC control group (untreated BM-MSC), recombinant human macrophage migration inhibitory factor (rhMIF) group and rhMIF+ISO-1 group. ELISA and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression level of IL-8 protein and mRNA in each BM-MSC group.Results:The expression levels of MIF protein in bone marrow fluid and plasma in AML group were (24.9±7.7) ng/ml and (60.5±12.1) ng/ml, the difference was statistically significant ( P < 0.01), and those in control group were (5.3±2.6) ng/ml and (2.0±1.1) ng/ml, respectively, and there were statistical differences between the two groups (t values were 136.71, 33.97 and 17.58, all P < 0.01). MIF protein expression levels in bone marrow fluid and plasma of AML patients in newly diagnosed group and PR+NR group were higher than those in CR group, and the differences were statistically significant (all P < 0.01). MIF protein expression levels were higher in bone marrow fluid and plasma of patients with ≥60 years of age, peripheral blood white blood cell count ≥30×10 9/L and bone marrow myeloblast ratio > 0.50 (all P < 0.05), but the differences were not statistically significant between patients with different gender (both P > 0.05). The detection results of each BM-MSC group showed that rhMIF promoted the IL-8 expression in BM-MSC at the gene and protein levels, which could be inhibited by the MIF inhibitor ISO-1 (all P < 0.01). Conclusion:The increased expression levels of MIF in bone marrow fluid and plasma of patients with AML are associated with the disease progression, and rhMIF can promote the expression of IL-8 in BM-MSC.

Objective To observe the clinical effect of treating Alzheimer's disease (AD) using transcranial magnetic stimulation.Methods One hundred and ninety-six patients with Alzheimer's disease were randomly divided into an observation group and a control group,each of 98.The observation group was given transcranial magnetic stimulation of the left and right dorsolateral frontal lobes of the brain and simultaneously given "8-shaped" coil stimulation.The stimulation intensity was 80％ of the motor threshold with a sequence of 2 s of stimulation at 5 Hz and 30 s rest for 30 min in each session.There were two sessions a day for 28 days.The control group was treated with identical pseudo-stimulation.Moreover,both groups were treated with intravenous injections of 20 ml of Ginkgo biloba extract dissolved in 250 ml of sodium chloride,or in the control group a glucose injection,one daily for two weeks.Before and after the treatment,the cognition,behavior and neuropsychological symptoms of both groups were evaluated using the mini mental state examination scale (MMSE),the AD rating scale (ADAS-cog),the activity of daily living (ADL) scale,a neuropsychiatric questionnaire (NPI) and an AD behavioral pathology rating scale (BEHAVE-AD) to compare the clinical effects.Results There were no significant differences in the groups' average scores on any of the evaluations before the treatment.After the treatment,the average MMSE and ADAS-cog scores in both groups had improved significantly,but with significantly greater improvement in the observation group.After the treatment,the average ADL,NPI and BEHAVE-AD scores of the observation group and the average ADL score of the control group were significantly lower than before the treatment.No significant differences were observed in the average NPI and BEHAVE-AD scores of the control group.After the intervention,the average ADL,NPI and BEHAVE-AD scores in the observation group were significantly lower than those of the control group.The total effectiveness rate of the observation group (90.8％) was significantly higher than that of the control group (62.2％).Conclusion Transcranial magnetic stimulation can significantly improve the cognitive,behavioral and neuropsychological status of patients with Alzheimer's disease.

OBJECTIVE:To study induction effect of euphornin on the apoptosis of cervical cancer Hela cells and its mechanism. METHODS:The cervical cancer Hela cells were divided into blank control group,cisplatin group(positive control, 10 mg/L) and euphornin low-dose,medium-dose and high-dose groups (50,100,200 mg/L). They were treated with relevant medicine. The inhibitory effect of Hela cells proliferation was tested by MTT assay after 24,48,72 h of medicine treatment. The apoptotic rate of Hela cells was measured by flow cytometry after 48 h of medicine treatment. Morphology of nucleus was detected by Hoechst 33258 staining. The protein expression of Cyt-C,Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9 and Caspase-10 were detected by Western blot assay. RESULTS:Compared with blank control group,inhibitory rate of cell proliferation and cell apoptosis rate were increased significantly in cisplatin group and euphornin groups(P＜0.05 or P＜0.01),and obvious staining, deformation,shrinking,fragmentation or apoptotic bodies was found in nucleus. Compared with blank control group,the protein expression levels of Cyt-C,Caspase-8 and Caspase-9 in euphornin low-dose,medium-dose and high-dose groups were increased significantly,while the protein expression level of Bcl-2 and Bcl-2/Bax ratio were decreased significantly(P＜0.05 or P＜0.01);the protein expression levels of Bax,Caspase-3 and Caspase-10 in euphornin medium-dose and high-dose groups were increased significantly(P＜0.05 or P＜0.01). CONCLUSIONS:Euphornin can significantly inhibit the proliferation of Hela cell and promote cell apoptosis,the effect of which will be achieved by activating the Caspase-dependent mitochondrion apoptosis pathway.

Objective To label granulocytes in a state of differentiation in mouse bone marrow (BM) with EdU (5-ethynyl-2′-deoxyuridine) for further understanding the changes in granulocyte produc-tion at different stages of differentiation during inflammation. Methods C57BL/6 mice were intraperitoneal-ly (i.p.) injected with EdU and heat-inactivated Escherichia coli(HI E.coli). BM cells were harvested at different time points after HI E.coli injection and then stained with fluorescent-conjugated antibodies(Abs). Myeloblasts,promyelocytes,myelocytes, metamyelocytes and band and segmented neutrophils were identi-fied by fluorescence-activated cell sorting(FACS). The percentage of EdU-positive cells in each population was recorded. Results The percentage of EdU-positive myeloblasts in mice increased by 10.0% at 24 h af-ter intraperitoneal injection with HI E.coli,but decreased by 75.0% and 23.0% at 48 h and 72 h,respec-tively. The percentage of EdU-positive promyelocytes declined by 23.0%,54.5%,64.3% and 77.8% at 24 h,48 h,72 h and 96 h,respectively. The percentage of EdU-positive myelocytes increased by 60.0% and 10.0% at 24 h and 48 h,but decreased by 80.0% and 90.0% at 72 h and 96 h. The percentage of EdU-positive metamyelocytes increased by 50.0% at 24 h,but decreased by 33.3%,61.5% and 66.7% at 48 h,72 h and 96 h. The percentage of EdU-positive band and segmented cells increased by 14.0% at 24 h,but decreased by 50.0%, 77.8% and 88.0% at 48 h, 72 h and 96 h. Conclusion Emergency granulopoiesis occurred 24 h after the establishment of HI E.coli-induced model of acute peritonitis, which meant that the proliferation of myeloid precursor cells,especially that of myelocytes and metamyelocytes,was accelerated and resulted in increasing number of mature neutrophils immigrating to sites of inflammation.