Objective: To explore the effects of porcine urinary bladder matrix (UBM) on the motility and polarization of bone marrow-derived macrophages in mice, so as to provide evidence for the rational selection of stent in clinical wound repair. Methods: The method of experimental research was used. The microstructure of porcine UBM and absorbable dressing was observed under scanning electron microscope. Polyacrylamide gel electrophoresis was used to observe the protein distribution of the two stent extracts. The primary macrophages were induced from bone marrow-derived cells isolated from six 6-8-week-old male C57BL/6J mice (mouse age, sex, and strain, the same below) and identified. Three batches of macrophages were divided into porcine UBM extract group and absorbable dressing extract group. The cells in each group were cultured with Dulbecco's modified Eagle medium/F12 medium containing the corresponding extracts. The cell migration rate was detected and calculated on 1, 3, and 7 d after scratching by scratch test. The number of migrated cells at 12 and 24 h of culture was detected by Transwell experiment. The percentages of CD206 and CD86 positive cells at 24 h of culture was detected by flow cytometer. The numbers of sample in the above cell experiments were all 3. An incision was prepared on the left and right back of twelve mice, respectively. The left incision of each mouse was included in porcine UBM group and the right incision was included in absorbable dressing group, and the corresponding stents were implanted into the incisions respectively. On post operation day (POD) 7 and 14, the number of inflammatory cells infiltrated in the stent was detected by hematoxylin-eosin staining; the number of F4/80, transforming growth factor-β₁ (TGF-β₁), vascular endothelial growth factor (VEGF), and matrix metalloprotein-9 (MMP-9) positive cells and type Ⅰ collagen deposition in stents were observed by immunohistochemistry; the percentages of F4/80, CD86, and CD206 positive cells were observed by immunofluorescence staining. The numbers of sample in the above animal experiments were all 6. Data were statistically analyzed with analysis of variance for factorial design, analysis of variance for repeated measurement, and independent sample t test. Results: Porcine UBM has a dense basement membrane structure on one side and porous propria containing a fibrous structures on the other. Both sides of the absorbable dressing had three-dimensional porous structure. In the molecular weight range of (50-70)×10³, multiple non-type Ⅰ collagen bands appeared in the lanes of porcine UBM extract, while no obvious bands appeared in the lanes of absorbable dressing extract. It had been identified that mouse bone marrow-derived cells had been successfully induced into macrophages. The cell migration rates in porcine UBM extract group were significantly higher than those in absorbable dressing extract group on 1, 3, and 7 d after scratching (with t values of 15.31, 19.76, and 20.58, respectively, P<0.05). The numbers of migrated cells in porcine UBM extract group were significantly more than those in absorbable dressing extract group at 12 and 24 h of culture (with t values of 12.20 and 33.26, respectively, P<0.05). At 24 h of culture, the percentage of CD86 positive cells in porcine UBM extract group ((1.27±0.19)%) was significantly lower than (7.34±0.14)% in absorbable dressing extract group (t=17.03, P<0.05);the percentage of CD206 positive cells in porcine UBM extract group was (73.4±0.7)%, significantly higher than (32.2±0.5)% in absorbable dressing extract group (t=119.10, P<0.05). On POD 7 and 14, the numbers of inflammatory cells infiltrated in the stents in porcine UBM group was significantly more than those in absorbable dressing group (with t values of 6.58 and 10.70, respectively, P<0.05). On POD 7 and 14, the numbers of F4/80, TGF-β₁, VEGF, and MMP-9 positive cells in the stents in porcine UBM group were significantly more than those in absorbable dressing group (with t values of 46.11, 40.69, 13.90, 14.15, 19.79, 32.93, 12.16, and 13.21, respectively, P<0.05); type Ⅰ collagen deposition in the stents in porcine UBM group was more pronounced than that in absorbable dressing group; the percentages of CD206 positive cells in the stents in porcine UBM group were significantly higher than those in absorbable dressing group (with t values of 5.05 and 4.13, respectively, P<0.05), while the percentages of CD86 positive cells were significantly lower than those in absorbable dressing group (with t values of 20.90 and 19.64, respectively, P<0.05), and more M2-type macrophages were seen in the stents in porcine UBM group and more M1-type macrophages were seen in the stents in absorbable dressing group. Conclusions: Porcine UBM can enhance macrophage motility, induce M2 polarization and paracrine function, create a microenvironment containing growth factors such as TGF-β₁ and MMP-9 tissue remodeling molecules, and promote tissue regeneration and extracellular matrix remodeling in mice.
Mice ; Male ; Animals ; Swine ; Vascular Endothelial Growth Factor A ; Urinary Bladder ; Matrix Metalloproteinase 9 ; Mice, Inbred C57BL ; Macrophages ; Collagen

OBJECTIVES: To develop a new Chinese medicine (CM)-based drug and to evaluate its safety and effect for suppressing acute respiratory distress syndrome (ARDS) in COVID-19 patients. METHODS: A putative ARDS-suppressing drug Keguan-1 was first developed and then evaluated by a randomized, controlled two-arm trial. The two arms of the trial consist of a control therapy (alpha interferon inhalation, 50 µg twice daily; and lopinavir/ritonavir, 400 and 100 mg twice daily, respectively) and a testing therapy (control therapy plus Keguan-1 19.4 g twice daily) by random number table at 1:1 ratio with 24 cases each group. After 2-week treatment, adverse events, time to fever resolution, ARDS development, and lung injury on newly diagnosed COVID-19 patients were assessed. RESULTS: An analysis of the data from the first 30 participants showed that the control arm and the testing arm did not exhibit any significant differences in terms of adverse events. Based on this result, the study was expanded to include a total of 48 participants (24 cases each arm). The results show that compared with the control arm, the testing arm exhibited a significant improvement in time to fever resolution (P=0.035), and a significant reduction in the development of ARDS (P=0.048). CONCLUSIONS Keguan-1-based integrative therapy was safe and superior to the standard therapy in suppressing the development of ARDS in COVID-19 patients. (Trial registration No. NCT04251871 at www.clinicaltrials.gov ).
Administration, Inhalation ; Adult ; China ; Coronavirus Infections ; diagnosis ; drug therapy ; mortality ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Follow-Up Studies ; Humans ; Integrative Medicine ; Interferon-alpha ; administration & dosage ; Lopinavir ; administration & dosage ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; diagnosis ; drug therapy ; mortality ; Risk Assessment ; Severe Acute Respiratory Syndrome ; diagnosis ; drug therapy ; mortality ; Severity of Illness Index ; Survival Rate

OBJECTIVE: To study the clinical features of coronavirus disease 2019 (COVID-19) in children aged <18 years. METHODS: A retrospective analysis was performed from the medical data of 23 children, aged from 3 months to 17 years and 8 months, who were diagnosed with COVID-19 in Jiangxi, China from January 21 to February 29, 2020. RESULTS: Of the 23 children with COVID-19, 17 had family aggregation. Three children (13%) had asymptomatic infection, 6 (26%) had mild type, and 14 (61%) had common type. Among these 23 children, 16 (70%) had fever, 11 (48%) had cough, 8 (35%) had fever and cough, and 8 (35%) had wet rales in the lungs. The period from disease onset or the first nucleic acid-positive detection of SARS-CoV-2 to the virus nucleic acid negative conversion was 6-24 days (median 12 days). Of the 23 children, 3 had a reduction in total leukocyte count, 2 had a reduction in lymphocytes, 2 had an increase in C-reactive protein, and 2 had an increase in D-dimer. Abnormal pulmonary CT findings were observed in 12 children, among whom 9 had patchy ground-glass opacities in both lungs. All 23 children received antiviral therapy and were recovered. CONCLUSIONS COVID-19 in children aged <18 years often occurs with family aggregation, with no specific clinical manifestation and laboratory examination results. Most of these children have mild symptoms and a good prognosis. Epidemiological history is of particular importance in the diagnosis of COVID-19 in children aged <18 years.
Adolescent ; Betacoronavirus ; Child ; Child, Preschool ; China ; Coronavirus Infections ; Humans ; Infant ; Pandemics ; Pneumonia, Viral ; Retrospective Studies

Objective The objective of this study was to explore the expression of miRNA-124(miR-124)and its correlation with E-cadherin and androgen receptor(AR)in triple-negative breast cancer(TNBC).Methods The expression of miR-124 was detected by RT-PCR in TNBC tissues and adjacent normal breast tissues.The expression of E-cadherin and AR in TNBC was detected by immunohistochemistry.Results The expression of miR-124 in TNBC tissues was significantly correlated with histological grade and the expression of E-cadherin(P<0.05),and the expression of miR-124 in TNBC tissues was significantly lower than that in adjacent normal breast tissues.No correlation was found between miR-124 and AR in TNBC tissues(P>0.05).Conclusion In TNBC,miR-124 may play an anti-tumor effect by modulating the expression of E-cadherin.

Objective To observe the therapeutic effect of Guizhi Decoction plus Yupingfeng Powder for chronic urticaria and its effect on serum total IgE level.Methods One hundred and twenty patients were randomly divided into treatment group,control group 1 and control group 2,40 cases in each group.The treatment group was treated with Guizhi Decoction,Yupingfeng Powder and Loratadine Tablets orally,the control group 1 was given intramuscular injection of Bacillus Calmette-Guerin polysaccharide nucleic acid and oral use of Loratadine Tablets,and the control group 2 was only given oral use of Loratadine Tablets.The course of treatment covered 5 weeks.The levels of serum total IgE in the three groups were determined before and after treatment,and the therapeutic effect was evaluated after treatment.Results (1) The total effective rate was 85.0％ in the treatment group,80.0％ in the control group 1,and 55.0％ in the control group 2.The effect of control group 1 and the treatment group was superior to that of the control group 2 (P＜ 0.05),but the difference was insignificant between the treatment group and control group 1 (P＞ 0.05).(2)Before treatment,the serum total IgE level of the three groups was higher than that of the normal control group (P ＜ 0.05).After treatment,the serum total IgE level of the three groups was obviously decreased (P ＜ 0.05 compared with that before treatment);the inter-group comparison results showed that the effect of the treatment group and control group 1 on decreasing the serum total IgE level was superior to that of the control group 2(P ＜ 0.05),while the effect of the treatment group was similar to that of the control group 1 (P ＞0.05).(3) During the treatment,blood routine examination indexes and hepato-renal function of the three groups showed no abnormal changes,neither adverse reaction was shown.Conclusion Guizhi Decoction plus Yupingfeng Powder exerts certain therapeutic effect for the treatment of chronic urticaria,and can decrease the serum total IgE level,thus to stabilize the curative effect and reduce the recurrence.

Carcinoma ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Nasopharyngeal Neoplasms ; radiotherapy ; Neoadjuvant Therapy ; methods ; Phytotherapy

Objective To construct an over-expressing plasmid containing full length of PELP1 gene and investigate the characterization of PELP1 expressions in different tissues.Methods Full length PELP1 gene was cloned using RT-PCR product of MCF-7 total RNA as the template.The PELP1 gene fragament was amplified by PCR and two enzyme sites EcoR I and Xho I were added.Then PELP1 gene was cloned into the pIRES2-EGFP palsimd (containing EcoR I and Xho I enzyme sites) to make reconstructed plasmid pIRES2-EGFP-PELP1.Enzyme digestion and sequencing were employed to assess the integrity and correctness of PELP1 gene cloned.The plasmid pIRES2-EGFP-PELP1 was transfected into human periodontal ligament stem cells,and the expression of PELP1 was examined by quantitative real-time-PCR or Western-blotting.Results The integrity and correctness of PELP1 gene were confirmed by digestion and sequencing.Conclusions The full length of PELP1 gene from MCF-7 cell was successfully cloned and over-expression plasmid pIRES2-EGFP-PELP1 constructed.

Objective To develop an electrochemical impedance immunosensor based on polypyrrole modified microelectrodes for periodontal bacteria rapid quantification.Methods Mcirofluidic chip with embedded IDAM was prepared by conventional photolithography process.Then polypyrrole structure was deposited on microelectrodes through electropolymerization method.Polypyrrole was biofuncationalized with mouse anti-Porphyromonas gingivalis gingipain K IgG antibody using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide-hydrochloride (EDC) chemistry.The linear relationship between the impedance and bacterial concentration,the impedance characteristic and the feasibility of the developed impedic immunosensor in quantifing Porphyromonas gingivalis was investigated respectively.Results The polypyrrole membrane structure was observed on work electrodes,and the lowest detection limit of the immunosensor was 107 cells/L in pure culture.In the concentration range of 107-1012 cells/L,a linear relationship was found between the normalized impedance change and the logarithmic value of the cell concentration.The total detection time from sampling to measurement was less than 1 h.Conclusions The immunosensor developed in the present study offered some insight into chair-side quantifing periodontal bacteria.

Objective To investigate the neointimal coverage at the very early phase after drugeluting stent (DES) implantation using optical coherence tomography (OCT).Methods OCT examination was performed immediately after stent deployment and about one week post stenting in 12 patients with coronary artery disease to detect neointimal coverage and stent thrombus.Sirolimus eluting stent implantation was also performed in 5 healthy Chinese minipigs,OCT and histopathology examination were made one week later in these minipigs.Results ( 1 ) Twenty-nine DES were implanted in 12 patients.There was no major cardiovascular event post stenting.The mean time of follow-up was ( 7.7± 2.6)d,the mean percentage of stent coverage was (21.8±17.7)％,and neointimal hyperplasia thickness was (42.9 ±32.2 ) μm and the percentage of malapposition struts was ( 1.5 ± 3.0) ％,respectively.Mural stent thrombus was found in 2 of the 12 patients (the percentage is 16.7％ ).(2) In the minipigs model,OCT evidenced that (43.2±11.5 ) ％ struts were covered by neointima with a mean neointimal hyperplasia thickness of ( 24.0 ± 8.5 ) μm at one week.Histopathology examination illustrated that the neointima was mainly consisted of proteoglycan,inflammation cells,fibrin and organized thrombus at the very early phase after DES implantation,while endothelial cells were barely found on the neointima.Conclusions Neointimal coverage is found as early as one week after DES implantation by OCT.The covered struts rate is very low and the main components of neointima are proteoglycan,inflammation cells,fibrin and organized thrombus.Re-endothelialization is rather poor at the very early phase post DES implantation.

OBJECTIVETo investigate the effect of insulin-like growth factor- I (IGF- I) on the proliferation and alkaline phosphatase (ALP) activity of human periodontal ligament cells (hPDLCs) under three-dimensional (3D) culture system.

METHODSThe hPDLCs were cultured from periodontium of human teeth by the outgrowth method. Rotary cell culture system (RCCS) was enrolled to set 3D culture system. Samples were set to four groups: Negative control group, positive control group (3D group, IGF-I group), and experimental group (3D with IGF- I group). Proliferation was tested with methylthiazolyl tetrazolium (MTT), and ALP activity was assayed by spectrophotometer at 1, 3, 5, 7 d respectively.

RESULTSCompared with that of negative control group, cell proliferation increased significantly in 3D with IGF-I group since 3 d (P < 0.05). Besides, the cell proliferation of 3D with IGF-I group was significantly higher than that of 3D group (P < 0.05). ALP activity of 3D with IGF- I group was significantly higher than that of negative control group, and 3D group at 3, 5, 7 d (P < 0.05).

CONCLUSIONIGF-I significantly promotes the proliferation and ALP activity of hPDLCs under 3D culture system.