Objective Hyperthyroidism and a diffuse goiter are the main symptoms of Graves' disease (GD) associated with autoantibodies to thyroid-stimulating hormone (TSH) receptor (TRAb). The present study was conducted to evaluate effects of autoantibodies in patients with GD (TRAb-IgG) on induction of the proliferation and c-fos mRNA expression in FRTL-5 cells (Fisher rat thyroid cell line).Methods Highly purified IgG fractions were isolated from 11 patients with GD, TRAb-IgG and 15 normal individuals (normal controls) with Protein A Sepharose CL-4B affinity column chromatograph. FRTL-5 cells, which had been grown to subconfluency and deprived of TSH for a few days. Then, these cells were used for measuring cAMP content, 3H-thymidine incorporation in cells and the expression of c-fos mRNA respectively.Results After stimulation of TRAb-IgG, the cAMP production and 3 H-thymidine incorporation in FRTL-5 cells were much higher than those from normal controls (P<0.05 respectively). Using 32P labelled v-fos probe by the Northern Blot method, the expression of c-fos mRNA could be induced by IgGs from patients with GD.Conclusions These data suggest that the stimulation of TRAb-IgG followed by cAMP production and 3H-thymidine incorporation is related to the induction of c-fos mRNA and, thus, to the growth of FRTL-5 cells.

Since the discovery of adipose-derived mesenchymal stem cell (ADSC) in more than ten years, a great progress has been made from its basic research to clinical application. Compared with bone marrow mesenchymal stem cells, ADSCs are more abundant in reserve, easier to obtain with fewer injuries and less complications. These cells have multiple differentiation potential and can differentiate into adipocytes, chondrocytes and osteoblasts with the influence of different inducing factors. Early studies of ADSCs mainly focused on the ability of multi-directional differentiation, espe-cially on the regeneration of bone defects and cartilage tissue. At present, the researches mainly focus on immunoregulation and paracrine function of ADSCs. Although ADSCs have made a great progress in clinical application, the cell preparation, use pattern, and mechanisms in clinical treatment are not clear. This paper elaborates on these issues.

Objective: To explore the effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells (hUCMSCs) and ciprofloxacin on Staphylococcus aureus (SA) in vitro. Methods: hUCMSCs were isolated from umbilical cord tissue of full-term healthy fetus after cesarean section and cultured. Cells in the third passage were used in the experiments after identification. SA strains isolated from wounds of burn patients in our burn wards were used in the experiments. Cells were divided into 0, 10, 100, and 1 000 ng/mL lipopolysaccharide (LPS) groups according to the random number table (the same dividing method below). Cells were cultured with culture medium of mesenchymal stem cells (MSCs) after being treated with medium containing the corresponding mass concentrations of LPS for 12 h. At post culture hour (PCH) 6, 12, and 24, 6 wells of culture supernatant of cells in each group were obtained to measure the content of LL-37 with enzyme-linked immunosorbent assay. Ninety blood agar plates were divided into ciprofloxacin control group (CC), ciprofloxacin+ supernatant group (CS), and ciprofloxacin+ supernatant+ LL-37 antibody group (CSL), with 30 blood agar plates in each group. Blood agar plates in group CC were coated with 1.5×10⁸ colony forming unit (CFU)/mL bacteria solution prepared with normal saline. Blood agar plates in group CS were coated with 1.5×10⁸ CFU/mL bacteria solution prepared with normal saline and culture supernatant of hUCMSCs (cultured by culture medium of MSCs, the same below) in double volume of normal saline. Blood agar plates in group CSL were coated with 1.5×10⁸ CFU/mL bacteria solution prepared with normal saline, culture supernatant of hUCMSCs in double volume of normal saline, and 2.6 μL LL-37 antibody in the concentration of 2 μg/mL. At PCH 12, 24, and 48, 10 blood agar plates of each group were harvested to observe the distribution of SA colony on blood agar plate and to measure the diameter of bacterial inhibition ring of ciprofloxacin. The minimum inhibitory concentration (MIC) of ciprofloxacin against SA of each group was recorded. Fractional inhibitory concentration (FIC) indexes of ciprofloxacin in groups CS and CSL at PCH 12, 24, and 48 were calculated, and the effect of synergy was evaluated. Data were processed with analysis of variance of factorial design, one-way analysis of variance, LSD-t test, Kruskal-Wallis test, and Mann-Whitney U test. Results: (1) At each PCH, the content of LL-37 in culture supernatant of cells in 10, 100, and 1 000 ng/mL LPS groups was higher than that in 0 ng/mL LPS group (with t values from 11.22 to 33.36, P values below 0.01); the content of LL-37 in culture supernatant of cells in 100 and 1 000 ng/mL LPS groups was higher than that in 10 ng/mL LPS group (with t values from 2.24 to 18.73, P<0.05 or P<0.01); the content of LL-37 in culture supernatant of cells in 1 000 ng/mL LPS group was higher than that in 100 ng/mL LPS group (with t values from 12.46 to 14.70, P values below 0.01). (2) At PCH 12, 24, and 48, the bacterial colonies in groups CC, CS, and CSL began to integrate over time. At PCH 12, 24, and 48, the diameters of bacterial inhibition ring of ciprofloxacin in group CC were 26, 24, and 23 mm, respectively, with no obvious change. At PCH 12, 24, and 48, the diameters of bacterial inhibition ring of ciprofloxacin in groups CS and CSL were 82, 71, 68 mm, and 74, 59, 56 mm, respectively, significantly longer than those of group CC. (3) At each PCH, the MIC of ciprofloxacin against SA was significantly higher in group CC than in groups CS and CSL (with Z values from 6.22 to 6.71, P values below 0.01); the MIC of ciprofloxacin against SA was significantly higher in group CSL than in group CS (with Z values all equal to 6.72, P values below 0.01). (4) FIC indexes of ciprofloxacin in groups CS and CSL at PCH 12, 24, and 48 were 0.011, 0.032, 0.032, and 0.122, 0.350, 0.350, respectively. The results indicated that culture supernatant of hUCMSCs had synergistically antibacterial effect on ciprofloxacin. Conclusions hUCMSCs can secrete LL-37, and the secretion level is increased with increase of LPS concentration. Combination of culture supernatant of hUCMSCs and ciprofloxacin can decrease the dosage of ciprofloxacin in resisting SA. Once LL-37 is neutralized, the synergistically antibacterial effect of culture supernatant of hUCMSCs is decreased.

Objective:To investigate the effects of human umbilical cord mesenchymal stem cell (hUCMSC) exosomes in the treatment of full-thickness skin defect wounds in mice through local wound application, subcutaneous injection at the wound margin, and tail vein injection, and to explore the optimal administration route of hUCMSC exosomes for wound treatment.Methods:This study was an experimental study. hUCMSC exosomes were extracted from the discarded umbilical cord tissue of three normal delivery women aged 25-35 years in the Department of Obstetrics and Gynecology of Baogang Hospital of Inner Mongolia and successfully identified. Totally 120 male BALB/c mice aged 6-8 weeks were selected, and full-thickness skin defect wounds were prepared on the back of them. According to the random number table, the injured mice were divided into control group (without drug administration), local wound application group, wound margin subcutaneous injection group, and tail vein injection group (with 30 mice in each group). Mice in the latter three groups were given 0.2 mL phosphate buffer solution containing 200 μg hUCMSC exosomes by local wound application, subcutaneous injection at the wound margin, and tail vein injection, respectively. On post injury day (PID) 7, 14, and 21, the general condition of the wound was observed, and the wound healing rate was calculated; the wound tissue was collected, the pathological changes and collagen fibers were observed respectively by hematoxylin-eosin staining and Masson staining, the number of new microvessels was observed by CD31 immunohistochemical staining, and the content of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) was detected by enzyme-linked immunosorbent assay. The sample number was 10 in each group at each time point.Results:On PID 7, 14, and 21, the wounds of mice in the 4 groups all healed gradually, and the wound healing of the mice in wound margin subcutaneous injection group was the best; the wound healing rates of mice in the three administration groups were significantly higher than those in control group ( P<0.05), the wound healing rates of mice in wound margin subcutaneous injection group and tail vein injection group were significantly higher than those in local wound application group ( P<0.05), and the wound healing rates of mice in wound margin subcutaneous injection group were significantly higher than those in tail vein injection group ( P<0.05). On PID 7, 14, and 21, the growth and epithelialization speed of the wound tissue of mice in the three administration groups were significantly accelerated, and the collagen fibers in the wounds of mice in the three administration groups were larger in number and more neatly arranged in comparison with the control group. On PID 7, 14, and 21, under every 200-fold visual field, the number of new microvessels in the wound tissue of mice in local wound application group was 24.1±2.5, 50.7±4.1, and 44.2±2.3, respectively, the number of new microvessels in the wound tissue of mice in wound margin subcutaneous injection group was 32.2±2.9, 67.5±4.9, and 53.6±3.7, respectively, and the number of new microvessels in the wound tissue of mice in tail vein injection group was 27.8±2.4, 59.1±3.7, and 49.6±2.6, respectively, which was significantly more than 20.6±1.7, 46.7±3.4, and 40.9±2.8 in control group ( P<0.05); the number of new microvessels in the wound tissue of mice in wound margin subcutaneous injection group and tail vein injection group was significantly more than that in local wound application group ( P<0.05); the number of new microvessels in the wound tissue of mice in wound margin subcutaneous injection group was significantly more than that in tail vein injection group ( P<0.05). On PID 7, 14, and 21, the content of TNF-α and IL-6 in the wound tissue of mice in the three administration groups was significantly less than that in control group ( P<0.05), the content of TNF-α and IL-6 in the wound tissue of mice in wound margin subcutaneous injection group and tail vein injection group was significantly less than that in local wound application group ( P<0.05), and the content of TNF-α and IL-6 in the wound tissue of mice in wound margin subcutaneous injection group was significantly less than that in tail vein injection group ( P<0.05). Conclusions:Local wound application, subcutaneous injection at the wound margin, and tail vein injection of hUCMSC exosomes can all promote the wound healing of full-thickness skin defects in mice through alleviating excessive inflammatory response and promoting angiogenesis. Among them, subcutaneous injection at the wound margin has a better therapeutic effect, indicating subcutaneous injection at the wound margin is the optimal administration route for hUCMSC exosomes in wound treatment.

The recombinant plasmid carrying the gene encoding 3C protease of Enterovirus 71 (EV71) was constructed, the recombinant protein was then expressed and purified, the functional activity was also measured. Firstly, the 3C protease gene was inserted into pET28a vector, the constructed recombinant plasmid was transformed into E. coli BL21 (DE3) for expression under the induction of IPTG. The expressed protein was purified by affinity chromatography (Ni-NTA) and the N-terminus His-tag was cleaved by enterokinase from 3C protease. The activity of 3C protease was evaluated with fluorescent peptide substrates. It was verified by restriction analysis and sequencing that recombinant plasmid pET28a-3C was constructed correctly and functionally expressed in E. coli BL21 (DE3) resulting in the production of recombinant 3C protease with a size of 22kD. Both His-tag and non-His-tag (cleaved by enterokinase) 3C protease exhibited similar enzyme activity to 3B-3C fluorescent peptide with Km, Vmax and Kcat values of 22 microM, 434nM. Min(-1) and 0.0669 Min(-1), respectively. The optimial pH and temperature were 7.0 and 30-37 degrees C, respectively. The acquirement of recombinant purified 3C protease with high activity has paved the way of further studies on anti-viral inhibitors, structural protein assembly, vaccine development and detection methods of EV71.
Cloning, Molecular ; Cysteine Endopeptidases ; chemistry ; genetics ; metabolism ; Enterovirus A, Human ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Kinetics ; Viral Proteins ; chemistry ; genetics ; metabolism

From January 2010 to December 2017, 4 patients of thumb with necrosis caused by electric burns (all male, aged from 31 to 58 years) were admitted to our hospital, with 1 patient of second degree injury of right thumb, 2 patients of third degree injury of right thumb, and 1 patient of third degree injury of left thumb. Routine debridement under general anesthesia was performed within 7 days after injury. The compound tissue flap of contralateral second toe was transplanted to reconstruct the thumb with third degree defect, and compound tissue flap of ipsilateral distal hallex was transplanted to reconstruct the thumb with second degree defect. Dorsalis pedics artery was anastomosed with radial artery, saphenous vein or dorsalis pedics vein was anastomosed with cephalic vein. The donor site was transplanted with split-thickness skin graft from autologous thigh. All the tissue flaps and skin grafts survived in 2 weeks after surgery. Within 1 year of follow-up, the reconstructed thumbs can achieve radial abduction and palmar abduction with good function. Reconstruction of thumb with free transplantation of compound tissue flap of toe is a good method to repair thumb with necrosis caused by electric burn.

Acute and chronic wounds seriously threaten patients' life health and quality of life, therefore, wound repair has become a hot topic of research for scholars at home and abroad in recent years. With the development of material science and tissue engineering, more and more biomaterials prepared from natural ingredients were used in basic research and clinical treatment of wound repair. Such biomaterials can be used as templates for wound tissue regeneration to induce autologous cell adhesion and migration, and promote the deposition of extracellular matrix, which have broad clinical application prospects. This paper reviews the characteristics and application advance of natural biomaterials which are popular in the field of wound repair, aiming to provide ideas for the research and development of new wound dressing and tissue engineering skin.

Objective:To establish and validate a risk prediction model of disseminated intravascular coagulation (DIC) by the screening independent risk factors for the occurrence of DIC in patients with electrical burns.Methods:The retrospective case series study was conducted. The clinical data of 218 electrical burn patients admitted to Baogang Hospital of Inner Mongolia from January 2015 to January 2023 who met the inclusion criteria were collected, including 198 males and 20 females, with the age of (38±14) years. The patients were divided into DIC group and non DIC group based on whether they were diagnosed with DIC during the treatment period. The following data of patients of two groups were collected and compared, including age, gender, total burn area, full-thickness burn area, injury voltage, whether osteofascial compartment syndrome occurred within 1 day after injury, duration of stay in burn intensive care unit, total length of hospital stay, whether combined with inhalation injury and multiple injuries, whether shock occurred upon admission, the abbreviated burn severity index score, and the acute physiology and chronic health evaluation Ⅱ score. The laboratory examination data of the patients within 24 hours after admission were also collected, including blood routine indexes: white blood cell count (WBC), hemoglobin level, platelet count (PLT), and neutrophil count; coagulation indexes: activated partial thromboplastin time (APTT), prothrombin time, thrombin time, and levels of D-dimer and fibrinogen (FIB); blood biochemistry indexes: aspartic transaminase, alanine transaminase, direct bilirubin, total bilirubin, total protein, albumin, blood glucose, creatinine, and urea nitrogen; blood gas analysis indexes: blood pH value, arterial partial pressure of oxygen, arterial partial pressure of carbon dioxide, bicarbonate, and base excess; and cardiac zymogram indexes: levels of myoglobin, troponin, lactate dehydrogenase, creatine kinase (CK), and α-hydroxybutyrate dehydrogenase. Data were statistically analyzed with chi-square test, Fisher's exact probability test, independent sample t test, and Mann-Whitney U test. For the variables with statistically significant differences in single factor analysis, the least absolute value selection and shrinkage operator (LASSO) regression was used to reduce the dimension, and the predictive factors for DIC in 218 patients with electrical burns were screened. The above-mentioned predictors were included in multivariate logistic regression analysis to find out the independent risk factors for DIC in 218 patients with electrical burns, and to draw the prediction model nomograms. The performance of the prediction model was evaluated by the receiver operating characteristic (ROC) curve and the area under the ROC curve, and the prediction model was validated by the calibration curve and clinical decision curve analysis (DCA). Results:Compared with those in non DIC group, the total burn area, full-thickness burn area, total length of hospital stay, and the proportions of high voltage caused injury, occurrence of osteofascial compartment syndrome within 1 day after injury, combination of inhalation injury, and occurrence of shock upon admission of patients in DIC group were significantly increased/prolonged (with Z values of -2.53, -4.65, and -2.10, respectively, with χ2 values of 11.46, 16.00, 7.98, and 18.93, respectively, P<0.05). Compared with those in non DIC group, the APTT, level of D-dimer, myoglobin, WBC, PLT, and levels of FIB, total bilirubin, and CK of patients within 24 hours after admission in DIC group were significantly prolonged/increased (with Z values of -2.02, -4.51, and -3.82, respectively, with t values of -3.84, -2.34, -2.77, -2.70, and -2.61, respectively), and the level of total protein and blood pH value were significantly reduced ( t=-2.85, Z=-2.03), P<0.05. LASSO regression analysis was carried out for the above 17 indicators with statistically significant differences. The results showed that injury voltage, the occurrence of shock upon admission, the occurrence of osteofascial compartment syndrome within 1 day after injury, and levels of D-dimer and total protein within 24 hours after admission were predictive factors for the occurrence of DIC in 218 patients with electrical burns (with regression coefficients of 0.24, 0.52, 0.35, 0.13, and -0.001, respectively). Multivariate logistic regression analysis showed that injury voltage, the occurrence of shock upon admission, the occurrence of osteofascial compartment syndrome within 1 day after injury, and D-dimer level within 24 hours after admission were independent risk factors for DIC in 218 patients with electrical burns (with odds ratios of 3.33, 4.24, 2.68, and 1.38, respectively, with 95% confidence intervals of 1.43-7.79, 1.78-10.07, 1.17-6.13, and 1.19-1.61, respectively, P<0.05). Based on the aforementioned four independent risk factors, the nomogram of prediction model for evaluating the probability of DIC in patients was drawn. The area under the ROC curve of prediction model was 0.88, and the 95% confidence interval was 0.82-0.95, indicating that the model had good predictive ability; the curve of prediction model tended to be near the ideal curve, indicating that the model had a high calibration degree; the clinical DCA of prediction model showed that the threshold probability of patients ranged from 4% to 97%, indicating that the model had good predictive ability. Conclusions:The injury voltage, the occurrence of shock upon admission, the occurrence of osteofascial compartment syndrome within 1 day after injury, and D-dimer level within 24 hours after admission are independent risk factors for the occurrence of DIC in patients with electrical burns. The prediction model established based on the above indicators can provide early warning for the occurrence of DIC in these patients.

OBJECTIVE: To observe the effect of acupuncture and moxibustion therapy of on ovulation and embryo implantation in luteal phase defect patients with spleen-kidney deficiency. METHODS: A total of 80 patients were randomly divided into an observation group and a control group, 40 cases in each one.In the observation group,acupuncture was applied at Shenting (GV 24), Shenque (CV 8), Guanyuan (CV 4), Qixue (KI 13), Lieque (LU 7), Gongsun (SP 4), Taixi (KI 3), Zusanli (ST 36) and Taichong (LR 3). And moxibustion was given at Taixi (KI 3) using moxibustion box during follicular phase, the stimulation of Taichong (LR 3) was strengthened during ovulatory phase, moxibustion was adopted at Shenque (CV 8) to Guanyuan (CV 4), Zusanli (ST 36) and Taixi (KI 3) during luteal phase. In the control group, acupuncture was applied at Guanyuan (CV 4), Dahe (KI 12), Sanyinjiao (SP 6), Ciliao (BL 32), Zhibian (BL 54) and Shenque (CV 8). Moxibustion was given at Sanyinjiao (SP 6) using moxibustion box during follicular phase, and moxibustion was adopted at Shenque (CV 8) to Guanyuan (CV 4) during luteal phase. The treatment were given every Monday, Wednesday and Friday, and the treatment were stoped during menstrual period in the two groups. Totally 3 menstrual cycle treatment were required, and 3 menstrual cycles were followed up. The pregnancy rate was observed after treatment, the ovulation rate, maximum folliclular diameter and difference of maximum folliclular diameters in ovulatory phase, serum progesterone (P) and basal body temperature (BBT) were compared before and after treatment in the two groups. RESULTS: In the observation group, 6 cases of successful pregnancy during treatment,10 cases in follow-up, the clinical pregnancy rate was 40.0% (16/40). In the control group, 1 case of successful pregnancy during treatment, 5 cases in follow-up, the clinical pregnancy rate was 15.0% (6/40). The clinical pregnancy rate in the observation group was higher than the control group (<0.05). The ovulation rate after treatment in the observation group was 90.0% (36/40), and the control group was 70.0% (28/40), compared before treatment, the ovulation rates were increased after treatment in the two groups (<0.05). The observation group was higher than the control group, but there was no significant difference between the two groups (>0.05). Compared before treatment, the maximum folliclular diameter and difference of maximum folliclular diameters in ovulatory phase, serum P after treatment were improved in the two groups (<0.05), and the improvements of the observation group were significant compared with the control group (<0.05). The BBT after treatment were superior to before treatment in the two groups (<0.05). After treatment, the normal BBT in the observation group was 33 cases, while the control group was 22 cases (<0.05). CONCLUSION Acupuncture and moxibustion therapy of can promote folliclar development, improve dominant follicle morphology, increase the level of serum P. The therapeutic effect is superior to routine acupuncture in increasing ovulation rate and improving pregnancy outcome.
Acupuncture Points ; Acupuncture Therapy ; Female ; Humans ; Infertility, Female ; therapy ; Luteal Phase ; Moxibustion ; Pregnancy ; Pregnancy Outcome

With the aging of population and the development of social economy, the incidence of chronic wounds is increasing day by day, while the incidence of burns and trauma remains at a high level, making wound repair an increasingly concerned area in clinical practice. Thymosin β4 is a naturally occurring small molecule protein in vivo, which is widely distributed in a variety of body fluids and cells, especially in platelets. Thymosin β4 has biological activities of promoting angiogenesis, anti-inflammation, anti-apoptosis, and anti-fibrosis, and has many important functions in wound repair. Thymosin β4 has been observed to promote the healing of various wounds, such as burns, diabetic ulcers, pressure ulcers. This paper will review the molecular structure, mechanism of wound healing promotion, pharmacokinetics, and clinical application of thymosin β4, aiming to introduce its potential in wound treatment and the shortcomings of current researches.
Burns/drug therapy* ; Humans ; Pressure Ulcer ; Thymosin/therapeutic use* ; Wound Healing/physiology*