Objective:To compare the efficacy and adverse reactions between intralesional betamethasone injection combined with botulinum toxin type A and topical hyaluronic acid and simple intralesional betamethasone injection and topical hyaluronic acid in treatment of keloid.Methods:A total of 58 patients with keloid were divided into two groups: intralesional betamethasone injection combined with botulinum toxin type A and topical hyaluronic acid group (combined group, n=28) and intralesional betamethasone injection combined with topical hyaluronic acid group (control group, n=30).In both two groups, the intralesional betamethasone injection was given once every 4 weeks for 3 times totally, and topical hyaluronic acid was applied every day.In combined group, an injection of botulinum toxin type A was given around the lesion after the first intralesional injection of betamethasone.The parameters for therapeutic efficacy and adverse reactions of the patients in two groups were recorded, and the clinical pictures were taken before and after each treatment, and they were analyzed and compared.Results:After 3 treatments, compared with control group, the lesion appearance in combined group was improved.In control group, the VAS value was reduced at 1 month, but was increased gradually at 2 and 3 months. In combined group, the VAS value was reduced gradually in 3 months. There was a significant difference in VAS value at 2 and 3 months between two groups (P<0.05).In combined group, the thickness of keloids were reduced gradually in 3 months, while the reduction was not obvious in control group (P<0.05).The recurrence of pain, itching and lesion height were observed 2 weeks after each treatment in control group, but there was no recurrence in combined group.The incidence rate of adverse reactions was 26.7% in control group, which had no significant difference compared with combined group(25.0%) (P>0.05).Conclusion:Compared with control group, the combined group shows better efficiency in the treatment of keloid and there is no increase of adverse reactions.The therapy is worthy of clinical application.

Objective:To study the expression characteristics of Dickkopf1 (DKK1) in different time and space during tooth development of the postnatal mice, and to provide the theoretical basis for clarifying the mechanism of Wnt signaling pathway in regulating the tooth development.Methods:The postnatal Kunming mice at days 0.5, 6.5, 12.5, 18.5, 24.5, and 30.5 respectively after birth were selected and divided into various groups by time,three in each group.The mice in each group were sacrificed and the paraffin sections of mandibular bone including the first molar were prepared at the thickness of 5 μm, followed by HE staining and immunohistochemical staining in order to detect the expressions of DKK1 in tooth tissue and periodontal tissue.Results:At 0.5 d after birth, the mandibular first molar tooth germ was in the bell stage.At 6.5 d the enamel development of mandibular first molar was almost completed, and the epithelium root sheath extended to the root direction.At 12.5 d the dentin development of crown was completed, with the root formatted about 1/3. At 18.5 d the root had formatted about 2/3.At 24.5 d the root had reached the full length.At 30.5 d the apical foramen was narrow, and the root development was basically completed.There was no DKK1 expression at 0.5 d, but it expressed in the odontoblasts and predentin at 6.5 d. From days 12.5 to 30.5,the expressions of DKK1 were positive in periodontal ligament, alveolar bone, and cellular cementum as odontoblasts, which were gradually increased with the prolongation of time.However, no expression of DKK1 was detected in the pulp.Conclusion:DKK1 shows regular expressions at different tooth developmental stages after birth, suggesting its potential role in the growth of dentin and periodontal tissues.

Objective:To investigate the application of lateral upper arm free flap in the repair of postoperative soft tissue defects after operation in the patients with tongue cancer, and to evaluate its survival rate and repair effect. Methods:Ten patients with tongue cancer, having a desire to repair soft tissue defect after tongue cancer operation, and their systemic state could tolerate the long time operation, were selected to repair tongue cancer postoperative defects with lateral upper arm flaps. Among them, 7 cases were male, 3 cases were female;aged 45-67 years old, average 52.1 years old.Radical resection of tongue cancer was primarily finished, then the flaps were designed and prepared to repair the defects of tongue according to the location and size of defects.After the operation, the survival rate of the flap was observed, and the effect of repair was evaluated according to the shape, size, volume and movement of the tongue.Results:The lateral upper arm flaps were stable and simple to prepare, the donor sites were concealed, and the upper limbs were not abnormal.All of flaps survived, and all of wounds healed primarily.The shape, size, and texture of tongues were satisfied, and the movements of tongue were not significantly restricted.After repair, the voice, mastication and swallowing function of the patients were good without obvious influence.Conclusion:The survival rate of lateral flap of upper arm is high, and the tongue shape and function were good after tongue defect repair.It is an ideal free flap for repairing the tongue tissue defect of tongue cancer after operation.

Objective:To detect the expressions of p16INK4a protein in the cervical lesion tissues of the Mongolian patients, and to explore the relationship between its expression and the occurrence and development of cervical cancer in the Mongolian patients.Methods:A total of 100 cases of paraffin sections of cervical cancer, cervical intraepithelial neoplasia(CIN),chronic cervicitis and uterine leiomyoma were divided into 25 cases of cervical cancer, 35 cases of CIN, 20 cases of chronic cervicitis, and 20 cases of uterine leiomyoma groups. The expressions of p16INK4a protein in different cervical tissues were detected by immunohistochemical SP method.Results:The positive rates of p16INK4a protern in cervical cancer, CIN, chronic cervicitis and uterine leiomyoma tissnes were 100.0%, 74.3%, 25.0%,and 10.0%, respectively.The results of K-W H rank sum test for multiple sample comparisons showed that the positive expression rate of p16INK4a protein in cervical cancer tissue was significantly higher than those in CIN, chronic cervicitis and uterine leiomyoma tissues(P<0.05).Conclusion:p16INK4a protein can be used as a indicator to screen the Mongolian patients with early cervical cancer.

Objective:To investigate the expression of fibrillin-1 (FBN-1) in the atrium tissue of the patients with rheumatic heart valve disease complicated with atrial fibrillation(AF), and to explore its relationship with atrial fibrosis in the patients with valvular atrial fibrillation.Methods:Eighty-four consecutive patients with rheumatic heart valve disease underwent cardiac surgery were enrolled in this study.The patients were divided into AF group(n=39) and sinus rhythm group(SR group, n=45).The clinical data of patients were collected before operation.The right atrium tissue (0.3-0.5 mm3) was disserted during operation.The degrees of right atrial fibrosis of the patients in two groups were observed by Masson staining.Western blotting method was used to measure the protein expressions of FBN-1 in atrium tissue of the patients in two groups.Results:There were no significant differences in the gender ratio, age, blood pressure, blood biochemical indicators and other aspects of medical history between two groups(P>0.05);the diameters of left and right atrium of the patients in AF group were significantly larger than those in SR group(P<0.05).The Masson staining results showed that there was obvious fibrosis in AF group, and the collagen volume fraction and collagen level in AF group were significantly higher than those in SR group (P<0.05).The expression level of FBN-1 in right atrium tissue in AF group was obviously higher than that in SR group(P<0.05).The expression level of FBN-1 protein in right atrium tissue of the patients with valvular atrial fibrillation was positively correlated with the collagen level(r=0.544,P=0.021).Conclusion:There is obvious atrium fibrosis in the patients with valvular atrial fibrillation;it is closely related to the up-regulation of the expression of FBN-1 gene.

Objective:To observe the protective effect of deproteinized extract of calf blood (DECB) on the kidney injury of the diabetic rats, and to discuss its mechanism preliminarily.Methods:The male Wistar rats were intraperitoneally injected with STZ (65 mg·kg-1) to establish the diabetes models, then the model rats were randomly divided into model (M)group, and metformin (MMet, 105 mg·kg-1) group , low dose of combined administration (ML,105 mg·k-1 metformin+94.5 mg·kg-1 ,DECB) group, medium dose of combined administration(MM, 105 mg·kg-1 metformin+ 189 mg·kg-1 DECB) group, high dose of combined administration(MH, 105 mg·kg-1 metformin +378 mg·kg-1,DECB) group, another ten Wistar rats were selected as normal control(NC)group.The rats were intragastrically administed with metformin and intraperitoneally injected with DECB, once a day, total of 8 weeks.The rats in NC group and M group were given normal saline solution.The weights and blood glucose levels of the rats in various groups were determined;the levels of serum high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), blood urea nitrogen (BUN), urinary albumin (UAlb), urine creatinine (UCR), serum creatinine (SCr), total cholesterol (TC), triglyceride (TG), uric acid (UA), glutathione (GSH), and malondialdehyde (MDA) were detected;the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined.The pathological changes of kidney tissue of the rats in various groups were detected by HE staining.Results:Compared with NC group, the weight of the rats in M group was reduced(P<0.05),and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG,and MDA were increased(P<0.05);the levels of HDL-C and GSH were obviously reduced(P<0.05),and the activities of SOD adn GSH-Px were obviously reduced(P<0.05).Compared with M group,the weights of the rats in MM and MH groups were increased (P<0.05), and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG, and MDA were decreased (P< 0.05);the levels of HDL-C and GSH were increased (P<0.05),and the activities of SOD and GSH-Px were increased (P<0.05).The pathological observation of kidney tissue showed that the rats in M group had obvios kidney tissue lesions with glomerular congestion and renal tubular edema compared with NC group;compared with M group,the pathological changes of the kidney tissue of the rats in drug administration groups were significantly improved, the renal tubular vacuoles were reduced, and the interstitial hyperplasia was not obvious.Conclusion:DECB combined with metformin can reduce the blood glucose level, regulate blood lipid, improve the pathological changes of kidney tissue in the diabetic rats, reduce the renal damage, and enhance the antioxidant capacity of the body.

Objective:To explore the effect of tectorigenin on myocardial fibrosis(MF) in the rats and clarify the related mechanism,and to provide reference for its clinical application. Methods:Sixty Wistar rats were randomly divided into normal control group,model group,positive drug (capropril) control group, and low,middle,high doses of tectorigenin groups(n=10).Except normal control group, the rats in other groups were used to construct MF models by subcutaneous injection of 5 mg·kg -1·d -1 isoproterenol (Iso) for 7 d.The rats in tectorigenin groups and captopril group were intragastricly administrated with different doses of tectorigenin (25,50,100 mg·kg-1·d-1)and captopril(10 mg·kg -1·d -1) from the second day after modeling for consecutive 28 d.Bl-420E+ biological function experiment system was used to detect the heart function;Heart mass index (HMI) and left ventricular mass index (LVMI) were measured after experiment.UV detection was used to measure the levels of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) in myocardial tissue.Microplate reader was used to measure the activities of lactic dehydrogenase(LDH) and creatine kinase(CK) and the levels of nitric oxide (NO)in serum.ELISA were used to detect the levels of collagen typeⅠ(ColⅠ) and collagen type Ⅲ (Col Ⅲ) in myocardium tissue of the rats.The pathological changes of myocardium tissue of the rats in various groups were observed by HE staining.Results:Compared with normal control group,the HR of rats in model groups was increased,and the left ventricular systolic pressure (LVSP) was decreased(P<0.01);the HMI and LVMI were increased(P<0.05),the levels of MDA in left ventricular myocardial tissue was increased(P<0.01),and the activity of SOD was decreased,the levels of serum ColⅠ,Col Ⅲ and the activities of LDH , CK were also increased(P<0.01);the level of NO in serum was decreased(P<0.01).Compared with model groups, the HR were decreased,LVSP were increased, and HMI and LVMI of the rats in different doses of tectorigenin groups were decreased in a dose-dependent manner;the levels of MDA were reduced;the activities of SOD were increased in myocardium tissue,and the CK activities and the ColⅠ and ColⅢ levels were decreased(P<0.05 or P<0.01);the LDH activities in middle and high doses of tectorgenin groups were decreased(P<0.01);and the levels of NO in serum in different doses of tectorigenin groups were significantly increased(P<0.05 or P<0.01) .Conclusion:Tectorigenin could inhibit the MF induced by Iso in the rats, and its mechanism may be related to antioxidation,scavenging free radical and inhibition of collagen synthesis.

Objective:To observe the repair effect of nano hydroxyapatite/magnesium(nHA-Mg)porous composite material modified by poly lactic acid/glycolic acid copolymer(PLGA) on the jaw bone defect of rabbits,and to elucidate the mechanisms preliminary.Methods:The mandibular defect models of 10 mm×5 mm×1 mm in the 18 rabbits were established.Nine rabbits were selected and implanted with nHA-Mg composite materials modified by PLGA in the left as experimental group and the right as blank control group;the other nine were implanted with nHA-Mg in the left as positive control group and the right as blank control group.The rabbits were sacrificed at 4, 8, 12 weeks (respectively 3 of experimental group and positive control group every time)and the mandibular defect areas were intercepted and observed by imaging and histological examination;the sizes of the newborn trabecula area and residual materials in experimental group and positive control group were compared.Results:Compared with positive control group and blank control group, the percentage of newborn trabecula area of the rabbits in experimental group was increased (P<0.05),and there was no statistic difference between positive control group and blank control group (P>0.05).The imaging results showed that the new bone formation can be observed in experimental group and better than positive control group and blank control group.The paraffin section results indicated that there were visible vascular tissue and newborn trabecula,the osteoblasts gathered around the bone trabecula;a lot of pits were located in the bone trabecula, and the pits contained osteocytes in experimental group.As the prolongation of time,thickened newborn trabecula, dense arrangement and trend of converting to lamellar bone were observed in experimental group.The hard tissue section results showed that the remaining amount of materials in experimental group was more than that in positive control group.Conclusion:nHA-Mg porous composite materials modified by PLGA can effectively reduce the rate of degradation in the body, promote osteogenesis and guide the bone regeneration.

Objective:To investigate the influence of P27RF-Rho mRNA gene silencing in the drug sensitivity of 5-fluorouracil(5-Fu)to the liver cancer SMMC cell line,and to provide theoretical basis for the treatment of advanced liver cancer.Methods:The P27RF-Rho RNAi vector was constructed and the P27RF-Rho gene silencing lentivirus were used to infect the SMMC7721 cells.Western blotting method was used to detect the gene silencing effect.The SMMC7721 cells were divided into Scramble-siRNA group, 5-Fu group, P27RF-Rho siRNA group and P27RF-Rho siRNA + 5-Fu group.Western blotting was used to detect the transfection efficiency of RNAi.MTT method was used to detect the cell growth in various groups.Scratching test was used to detect the migration ability of cells in various groups.Transwell experiment were used to detect the invasion ability of cells in various groups.The expressions of P27 and RhoC protein were detected by Western blotting method.Results:P27RF-Rho RNAi lentiviral vector was successfully constructed.The Western blotting results showed that the expression of P27RF-Rho protein in P27RF-Rho siRNA group was decreased compared with 5-Fu group and Scramble-siRNA group(P<0.05).Compared with other three groups, the growth speed of the cells in P27RF-Rho siRNA + 5-Fu group was significantly decreased(P<0.05).The migration ability of the cells in P27RF-Rho siRNA + 5-Fu group was significantly lower than those in other three groups (P<0.01);the average number of cells passing through the Transwell microporous membrane was significantly less than those in other three groups (P<0.01).The Western blotting analysis results showed that the expression level of P27 protein in the cells in P27RF-Rho siRNA + 5-Fu group was significantly higher than those in other three groups(P<0.05);the expression level of RhoC protein was significantly lower than those in other three groups(P<0.05).Conclusion:P27RF-Rho gene silencing can significantly enhance the drug sensitivity of 5-Fu to SMMC7721 cells.

Objective:To investigate the influence of multilayer alginate chitosan sustained-release microspheres loading vascular endothelial growth factor(VEGF) and vancomycin in the proliferation and osteogenic differentiation of human plaacenta-derived mesen chymal stem cells(HPMSCs),and to provide theoretical basis for its clinical application in the repair of bone defect.Methods:The microspheres were prepared based on the previous research and HPMSCs were co-cultured with drug (VEGF/vancomycin)-loaded microspheres (drug-loaded microspheres+HPMSCs group), non-drug loaded microspheres (microspheres+HPMSCs group) and without any microspheres (HPMSCs group).Then the proliferation rate of HPMSCs was identified by CCK-8 kit.The osteogenic differentiation potential of HPMSCs was detected by Alizarin red staining and alkaline phosphatase (ALP) kit when the HPMSCs had been co-cultured with drug loaded microspheres in osteogenic medium (HPMSCs+drug-loaded microspheres+induction group), non-drug loaded microspheres in osteogenic medium (HPMSCs+microspheres+induction group), without any microspheres in osteogenic medium (HPMSCs+induction group) and without any micropheres in normal medium (HPMSCs+PBS group) for 21 d.Results:Compared with HPMSCs group,the proliferation rates of HPMSCs in drug-loaded microspheres+HPMSCs and microspheres+HPMSCs groups had no significant changes (P>0.05).The calcium deposition in HPMSCs+drug-loaded microspheres+induction group was more than those in microspheress+HPMSCs+indution group and HPMSCs+induction group after Aalizarin red staining;the ALP activity in drug-loaded microspheres+HPMSCs+indution group was higher than those in microspheres+HPMSCs+indution group and HPMSCs+induction group (P<0.05),and the ALP activity in microspheres+HPMSCs+induction group was higher than that in HPMSCs+induction group(P<0.05).Conclusion:The sustained-release microspheres loading VEGF and vancomycin have no significant effect on the proliferation activity of HPMSCs and the microspheres could stimulate the osteogenic differentiation of HPMSCs.