OBJECTIVETo investigate the possibility of placenta mesenchymal stem cells (PMSCs) differentiation into dermal fibroblast, and the potency of PMSCs used in cutaneous wound healing and stored as seed cells.

METHODSEnzyme digestion method was used to obtain PMSCs, and PMSCs were amplified after culture in vitro. Flow cytometry assay, osteogenic and adipogenic differentiation were done for MSCs identification. The induction medium composed of DMEM/F12 + 50 microg/ml VC + 100 ng/ml connective tissue growth factor (CTGF) was added into the 24-well plate for 16 days induction period. Pictures were taken to record morphologic change. Immunofluorescence tests were performed to detect Vimentin, FSP-1, collagen I , collagen III, desmin and laminin expression before and after induction. At the same time osteogenic and adipogenic differentiation were used to assay the differentiation ability change after induction. The induced dermal fibroblasts were frozen in liquid nitrogen and recovery and trypan blue was used to detect cell viability.

RESULTSAfter CTGF induction, PMSCs got obvious fibroblasts morphology, the protein level of Vimentin, FSP-1, collagen I, collagen III and Laminin increased, PMSCs started to express Desmin, the dermal fibroblasts specific proteins, and osteogenic and adipogenic differentiation ability was diminished. PMSCs were successfully induced into dermal fibroblasts, and these induced cells could get a high cell viability ( more than 90% ) after recovery.

CONCLUSIONSPMSCs could be induced into dermal fibroblasts by CTGF in vitro. PMSCs have the potential application in skin wound healing, and can be used as seed cells of dermal fibroblasts.

Cell Differentiation ; drug effects ; Cells, Cultured ; Connective Tissue Growth Factor ; pharmacology ; Female ; Fibroblasts ; cytology ; drug effects ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Placenta ; cytology ; Pregnancy

The purpose of study was to investigate the feasibility of the application of cationic propyl gallate (C-PG) as inducer of platelet aggregation for evaluating the platelet function of single-donor plateletpheresis and identifying the incidence of defective platelet function among donors. Experiments were as follows: 3 healthy volunteers' platelet aggregation induced by 100-300 micromol/L C-PG was determined by LG-PABER analyzer to observe the effect of C-PG concentration on platelet aggregation; 30 healthy volunteers' platelet aggregation before and 24 hours after administration of 200-400 mg acetylsalicylic acid (ASA) was examined after induction by 200 micromol/L C-PG for determining the cut-off value to discriminate platelet dysfunction donors; the platelet aggregation of 483 platelet donors was detected and the activated plasma clotting time (APCT) of donors who have deficiency in platelet aggregation was examined for investigating the incidence of defective platelet function among donors. The results showed that platelets were activated by C-PG induction in a dose dependent manner, when concentration of C-PG reached 200 micromol/L, the percentage of platelet aggregation was highest. It significantly decreased after 24 hours with ASA than that before the administration (P < 0.001), especially in 180 seconds induced by C-PG. If cut-off point was fixed on the platelet aggregation < 20% in 180 seconds, donors of platelet dysfunction can be selected effectively. 25 of defective platelet aggregation function among 483 donors were detected, and 11 out of 25 platelet dysfunction donors had the deficiency in procoagulant activity with prolonged APCT. It is concluded that C-PG as inducer of platelet aggregation is feasible to screen the platelet function of donors. Five percent of platelet donors has function defect examined by C-PG as inducer of platelet aggregation.
Antioxidants ; chemistry ; pharmacology ; Aspirin ; administration & dosage ; Blood Donors ; Blood Platelets ; cytology ; drug effects ; physiology ; Cations ; chemistry ; Humans ; Platelet Activation ; drug effects ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; administration & dosage ; Platelet Function Tests ; Platelet Transfusion ; Propyl Gallate ; administration & dosage ; chemistry ; Whole Blood Coagulation Time

BACKGROUND:Three-dimensional (3D) printing technology is currently one of the most advanced industrial manufacturing technologies. The surgical template prepared based on the 3D printing technology is mainly made of resin, and a great improvement in its accuracy is required. However, the clinical application of the surgical template made of metal is rarely reported. OBJECTIVE:To evaluate the clinical value of 3D-printing implant template in the restoration of free-end missing teeth.METHODS:A prospective study was conducted in 64 enrolled patients with free end-tooth defects. All the patients were randomly assigned to receive traditional implant template (control group,n=32) or 3D-printing implant template (study group,n=32), and 3-6 months later, the patients were subjected to crown restoration. At 6 months after crown restoration, cone beam computed tomography was performed to compare the deviation of the implant tip and neck (including vertical, buccolingual, mesial-distal). Success rate and chewing rate were compared between the two groups at 6 months after crown restoration; patient satisfaction assessment was done and compared between the two groups at 1 year after crown restoration. RESULTS AND CONCLUSION:There were no significant differences between the two groups in the success rate and chewing rate (98.7％ vs. 95.6％; 97.4％ vs. 97.1％,P>0.05). The vertical, buccolingual, mesial-distal deviations of the implant tip were significantly lower in the study group than the control group (P<0.05), while there was no difference in the vertical and buccolingual deviations of the implant neck between the two groups (P>0.05), and the mesial-distal deviation of the implant neck was significantly lower in the study group than the control group (P<0.05). In addition, there was no difference in the patient satisfaction between the study and control groups (94％vs. 91％, P>0.05). To conclude, the 3D printing implant template can effectively reduce implant excursion based on the assurance of therapeutic efficacy and patient satisfaction, which is of great significance in the restoration of free-end tooth loss.

The objective of this study was to investigate the serologic characters of enzyme-only red cell antibody and its clinical significance, and to provide basis for the safety of blood transfusion. The patient serum containing enzyme-only antibody was used to react with the red cells of donors, panel cells and auto-cells in various medium. Absorption and elution test were also per formed. The results showed that this blood sample was found to contain an antibody that reacted with donor red cells and panel cells only in papain medium, but was not demonstrable by indirect antiglobulin test and other method s. Decline of antibody titers was observed after absorption test, but antibody activity was not detected in the elute. The patient underwent transfusion with 600 ml of Rh type identical RBCs, without any hemolytic transfusion reaction. In conclusion, enzyme-only antibody usually doe s not lead to hemolytic transfusion reaction.
Aged ; Aged, 80 and over ; Erythrocytes ; immunology ; Hemolysis ; Humans ; Isoantibodies ; immunology ; Male ; Papain ; pharmacology ; Transfusion Reaction

OBJECTIVETo study the effect of cigarette smoke on the sexual function of male rats.

METHODSBased on Ozyurt's smoking model, we equally divided 30 male adult Sprague-Dawley rats into a control and a smoking group, and exposed the latter to cigarette smoke for 60 days. A week before the end of the experiment, we added 5 female rats to each group and observed their mating through 24-hour video surveillance. Sixty days later, all the rats were killed for the determination of the level of testosterone (T) in the plasma and the activity of nitric oxide synthase (NOS) in the corpus cavernosum, and Masson-dyeing image analysis of the penile tissue.

RESULTSCompared with the controls, the rats in the smoking group showed a significant reduction in the times of mating, the level of plasma T (P < 0.05) and the activity of NOS in the penile cavernous tissue (P < 0.05), but a slight increase in the collagen fibers and obvious changes in the blood sinuses.

CONCLUSIONCigarette smoke seriously affected penile erection in the experimental rats. The decrease in plasma T, NOS activity and the area of smooth muscle may be an important mechanism underlying their erectile dysfunction.

Animals ; Erectile Dysfunction ; Male ; Muscle, Smooth ; metabolism ; Nitric Oxide Synthase ; metabolism ; Penile Erection ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Smoke ; adverse effects ; Testosterone ; blood ; Tobacco ; adverse effects

OBJECTIVETo identify molecular markers of lung squamous cell carcinoma by cDNA microarray technique.

METHODScDNA expression profiles were examined by microarrays of 6 surgical specimens of stage I lung squamous cell carcinomas. Those genes, either up-regulated or down-regulated in every specimen studied, were identified. The expression levels of nm23 and BRCA2 by the squamous cell carcinoma of the lung were further examined by immunohistochemical techniques.

RESULTSA total of 107 genes were identified, of which 26 were up-regulated and 81 were down-regulated in all six specimens. Immunohistochemical staining showed that, compared with normal lung tissues, the intensity of nm23 expression by the squamous cell carcinoma of lung was significantly increased while that of BRCA-2 was decreased.

CONCLUSIONcDNA microarrays can be used to identify gene expression profile of lung cancer, some of which may be used as markers of lung squamous cell carcinoma.

BRCA2 Protein ; metabolism ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; genetics ; metabolism ; Male ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; metabolism ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo analyze the phenotype and genotype in three Chinese pedigrees with inherited dysfibrinogenemia.

METHODSLaboratory tests including activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), reptilase time (RT), and the activities of antithrombin (AT:C), protein C (PC:C) and protein S(PS:C) were detected in three pedigrees. The activity and antigen of plasma fibrinogen (Fg) were analyzed by Clauss and immunoturbidimetry methods, respectively. The Fg of three probands was assessed by Western blot and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The sequences of all the exons and exon-intron boundaries of the three Fg genes FGA, GFB and FGG were amplified by PCR and analyzed by direct sequencing.

RESULTSThree probands had normal APTT, PT, PC:C, PS:C and AT:C, but prolonged TT and RT. The activity levels of the 3 probands's plasma Fg were reduced, but antigen levels were normal. Western blot and SDS-PAGE showed no abnormal molecular weight of Fg. The 3 heterozygous mutations of γ Arg275His, Aα Pro18Leu and Aα Arg16Cys were identified in the 3 probands, respectively.

CONCLUSIONThe three probands with dysfibrinogenemia were caused by the mutations of γ Arg275His, Aα Pro18Leu and Aα Arg16Cys, respectively. Both Aα Pro18Leu and Aα Arg16Cys were first reported in Chinese population.

Adult ; Afibrinogenemia ; genetics ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Female ; Fibrinogen ; genetics ; Genotype ; Humans ; Middle Aged ; Mutation, Missense ; Pedigree ; Phenotype

Objective To explore the diagnostic value of non-real-time radial probe endobronchial ultrasound (RP-EBUS) guided transbronchial lung biopsy (TBLB) for peripheral lung cancer and analysis of false negative results. Methods A retrospective analysis of the clinical and imaging data of 256 patients with peripheral lung cancer between March 2013 and December 2016, all the cases underwent non-real-time RP-EBUS guided TBLB, then evaluate its significance in the diagnosis of peripheral lung cancer and analyze the reasons of false negative results. Result In 256 patients who received non-real-time RP-EBUS examinations, 73.83% (189/256) of peripheral lung cancer were detected by RP-EBUS and the positive rate of RP-EBUS guided TBLB was 61.33% (157/256). The positive rate of non-real-time RP-EBUS guided TBLB was correlated with lesions >2 cm in diameter, lesions close to visceral pleura, ultrasonic image characteristics and the RP-EBUS probe surrounding by lesion (P < 0.05). The positive rate of non-real-time RP-EBUS guided TBLB was not correlated with RP-EBUS probe passed through lesions and times of biopsy (P > 0.05). Complications including bleeding, chest pain and pneumothorax recovered spontaneously. Conclusion Non-real-time RP-EBUS guided TBLB was a practical technology for diagnosis of peripheral lung cancer with high diagnostic rate and good safety. Lesion size, connection to visceral pleura, ultrasonic image characteristics and the RP-EBUS probe surrounding by lesion influenced the diagnostic yield. Improvement of operative skills can reduce false negative results.

OBJECTIVE: To study the cytokine patterns in patients with rheumatoid arthritis (RA) and healthy individuals and identify candidate serum biomarkers for clinical diagnosis of RA. METHODS: This study was conducted among 59 patients diagnosed with RA in our hospital from 2015 to 2019 with 46 age- and gender-matched healthy subjects who received regular physical examinations in our hospital as the control group. Serological autoimmune profiles of 5 RA patients and 5 healthy control subjects were obtained from human cytokine microarrays. We selected 4 differentially expressed cytokines (LIMPII, ROBO3, Periostin and IGFBP-4) and 2 soluble cytokine receptors of interest (2B4 and Tie-2) and examined their serum levels using enzyme-linked immunosorbent assay in 54 RA patients and 41 healthy control subjects. Spearman correlation test was performed to assess the correlation of serum cytokine and soluble receptor expression levels with the clinical features including rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), disease activity score (DAS28) and health assessment questionnaire (HAQ). Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic capability of these cytokines. RESULTS: We identified 6 dysregulated cytokines and soluble receptors (2B4, LIMPII, Tie-2, ROBO3, periostin and IGFBP-4) in RA patients (P < 0.01). The serum levels of LIMPII, ROBO3 and periostin were significantly correlated with the disease activity indicators including RF (P < 0.001), CRP (P < 0.001), DAS28 (P < 0.001) and HAQ (P < 0.001) in RA patients. Among the 6 candidate cytokines, 2B4 showed the largest area under the curve (AUC) of 0.861 for RA diagnosis (P < 0.001), followed then by LIMPII, ROBO3, periostin, Tie-2 and IGFBP-4. CONCLUSION Serum levels of LIMPII, ROBO3 and periostin can be indicative of the disease activity of RA, and serum 2B4, LIMPII, periostin, ROBO3, IGFBP-4 and Tie-2 levels may serve as biomarkers for the diagnosis of RA.
Arthritis, Rheumatoid/diagnosis* ; Biomarkers ; C-Reactive Protein ; Cytokines ; Humans ; Insulin-Like Growth Factor Binding Protein 4 ; Protein Array Analysis ; Receptors, Cell Surface

BACKGROUNDFetal brain development is a complicated process that continues throughout pregnancy. Fetal sulcus development has typical morphological features. Assessment of fetal sulcus development to understand the cortical maturation and development by prenatal ultrasound has become widespread. This study aimed to explore a reliable method to assess cortical sulcus and to describe the normal sonographic features of cortical sulcus development in the human fetus between 18 and 41 weeks of gestation.

METHODSA cross-sectional study was designed to examine the fetal cortical sulcus development at 18-41 weeks of gestation. Ultrasound was used to examine the insula, sylvian fissure (SF), parieto-occipital fissure (POF), and calcarine fissure (CF). Bland-Altman plots were used for assessing the concordance, and the intraclass correlation coefficient was used for assessing the reliability.

RESULTSSF images were successfully obtained in 100% of participants at 22 weeks of gestation, while the POF images and CF images could be obtained in 100% at 23 weeks of gestation and 24 weeks of gestation, respectively. The SF width, temporal lobe depth, POF depth, and the CF depth increased with the developed gestation. The width of uncovered insula and the POF angle decreased with the developed gestation. By 23 weeks of gestation, the insula was beginning to be covered. Moreover, it completed at 35 weeks of gestation. The intra- and inter-observer agreements showed consistent reproducibility.

CONCLUSIONSThis study defined standard views of the fetal sulcus as well as the normal reference ranges of these sulcus measurements between 18 and 41 weeks of gestation. Such ultrasonographic measurements could be used to identify fetuses at risk of fetal neurological structural disorders.

Adult ; Brain ; embryology ; Cross-Sectional Studies ; Female ; Fetal Development ; physiology ; Fetus ; embryology ; Gestational Age ; Humans ; Pregnancy ; Ultrasonography, Prenatal ; methods