ObjectiveTo explore the mechanism of Cangxi Tongbi capsules （CXTB） in regulating the p38 mitogen-activated protein kinase （p38 MAPK）/NOD-like receptor protein 3 （NLRP3）/cysteine protease-1 （Caspase-1） signaling pathway to inhibit pyroptosis of cartilage cells in knee osteoarthritis （KOA）. MethodSixty male SD rats were randomly divided into a sham operation group， a model group， low， medium， and high dose CXTB groups， and a positive control group， with 10 rats per group. The modified Hulth method was employed to establish a rat model of KOA. According to their respective assignments， rats were administered CXTB （0.25， 0.5， 1.0 g·kg^-1） and Celecoxib （24 mg·kg^-1） by gavage. The sham operation and model groups were given an equivalent volume of physiological saline. Treatment was performed once daily for 28 days. Micro-computed tomography （Micro-CT） was used to assess bone volume/total volume （BV/TV） and trabecular separation （Tb.Sp）. Joint degeneration was evaluated through hematoxylin-eosin （HE） staining， safranin-fast green （SO） staining， and Osteoarthritis Research Society International （OARSI） scoring. Western blot analysis was conducted to measure the expression levels of p38 MAPK， phosphorylated p38 MAPK （p-p38 MAPK）， NLRP3， Caspase-1， and gasdermin D （GSDMD） proteins. Real-time PCR was used to assess mRNA expression levels of p38 MAPK， NLRP3， Caspase-1， and GSDMD genes. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum concentrations of inflammatory cytokines including tumor necrosis factor-alpha （TNF-α）， interleukin-1 beta （IL-1β）， and interleukin-18 （IL-18）. After knee replacement surgery， cartilage tissue was analyzed using Western blot to assess the protein expression levels of p38 MAPK， p-p38 MAPK， NLRP3， Caspase-1， and GSDMD， and Real-time PCR was used to evaluate gene expression levels of p38 MAPK， NLRP3， Caspase-1， and GSDMD. ResultMicro-CT analysis revealed significant narrowing of the joint space and increased bone spur formation in KOA rats compared with the sham operation group， with a decrease in BV/TV ratio and an increase in Tb.Sp value （P<0.01）. Serum levels of TNF-α， IL-1β， and IL-18 were elevated （P<0.01）. The protein expression levels of p-p38 MAPK， NLRP3， Caspase-1， and GSDMD in cartilage were significantly increased （P<0.01）， and the mRNA expression levels of p38 MAPK， NLRP3， Caspase-1， and GSDMD were also enhanced （P<0.01）. Significant differences in protein expression of p-p38 MAPK， NLRP3， Caspase-1， and GSDMD were observed between normal and diseased cartilage tissues after knee replacement surgery （P<0.05）， and the gene expression of p38 MAPK， NLRP3， Caspase-1， and GSDMD were also significantly different （P<0.01）. HE and SO staining showed roughened joint surfaces， reduced cartilage thickness， and disordered cellular arrangement in KOA rats. OARSI scores were significantly higher （P<0.01）. Compared with the model group， treatment with low， medium， and high concentrations of CXTB resulted in increased BV/TV ratios and decreased Tb.Sp values in the knee joints of rats （P<0.01）. HE and SO staining indicated a trend towards smoother joint surfaces and reduced OARSI scores （P<0.01）. The protein expression levels of p-p38 MAPK， NLRP3， Caspase-1， and GSDMD were notably decreased （P<0.05）， as were the mRNA expression levels of p38 MAPK， NLRP3， Caspase-1， and GSDMD （P<0.01）. Additionally， serum concentrations of TNF-α， IL-1β， and IL-18 were significantly reduced （P<0.01）. ConclusionCXTB intervention may alleviate knee joint degeneration in KOA rats and inhibit the expression of inflammatory factors and pyroptosis of cartilage cells， thereby protecting cartilage. The underlying mechanism may involve modulation of the p38 MAPK/NLRP3/Caspase-1 signaling pathway.

Objective to investigate the clinical effect of constant temperature vaginal mould in preventing vaginal stricture in patients with cervical cancer treated with radiotherapy. Methods from January 2017 to December 2018, 80 patients with cervical cancer were selected and divided into vaginal irrigation control group (n = 40) and observation group (n = 40). The incidence of vaginal stricture was compared between the two groups. Results the incidence of vaginal stenosis was 32.50% in the observation group and 70.00% in the control group (P < 0.05). Conclusion vaginal mould is helpful to reduce the incidence of vaginal stenosis caused by external irradiation and close irradiation, which is worth popularizing in the future.

Objective: To explore the relative risk factors, clinical intervention and prognosis of hemorrhagic cystitis (HC) in patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: From January 1 2010 to May 31 2017, 425 patients with allo-HSCT received a retrospective analysis. Results: ①Among the 425 patients, 262 were male and 163 were female. The median age was 26 (2-56) years old. There were 138 cases of acute myeloid leukemia (AML) , 96 cases of acute lymphoblastic leukemia (ALL) , 29 cases of myelodysplastic syndrome (MDS) , 98 cases of severe aplastic anemia (SAA) and 64 cases of chronic myeloid leukemia (CML) . 221 cases of sibling match transplantation, 89 cases of unrelated donor transplantation and 115 cases of haplotype transplantation. ②108 patients (25.41%) developed HC, with the median time of onset of 32 (3-243) days and the median duration of 20 (3-93) days; 33 cases (30.56%) were grade Ⅰ, 49 cases of grade Ⅱ (45.36%) , 21 cases (19.44%) of grade Ⅲ, and 5 cases (4.63%) of grade Ⅳ. ③103 cases of HC were cured, 5 patients were ineffective, 12 patients died and died of transplantation related complications (infection, recurrence, severe acute GVHD, secondary implant failure) . ④Univariate analysis showed that age < 30, type of transplantation, CMV and acute GVHD were associated with the occurrence of HC after allo-HSCT. Multivariate analysis showed that acute GVHD was an independent risk factor for HC after allo-HSCT. Conclusion Prognosis of HC after allo-HSCT was better after timely treatment.

Objective: To investigate and analyze the impact on PLT recovery of recombinant human thrombopoietin (rhTPO) in severe aplastic anemia (SAA) patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: A retrospective analysis of Hematology Division of General Hospital of Jinan Military Command was conducted in the 85 SAA cases who treated with allo-HSCT from January 2010 to March 2017. According to the administration of medicines for platelets, 85 patients were divided into rhTPO group (n=29), rhIL-11 group (n=27) and blank group (n=29), respectively. The median time of PLT ≥20×10⁹/L, PLT ≥50×10⁹/L, and PLT ≥100×10⁹/L, the numbers of megakaryocytes in marrow smear (25±5) days after transplantation and the quantities of platelet transfusion were analyzed retrospectively. The adverse events of rhTPO and rhIL-11 groups were observed. Results: There were no significant differences in the recovery of granulocytes and PLT ≥20×10⁹/L among the three groups (P>0.05). The time of PLT ≥50×10⁹/L in rhTPO group was shorter than that in blank group [16.5 (11-39) d vs 22 (14-66) d, P<0.05], as well as the time of PLT ≥100×10⁹/L [rhTPO: 23 (12-51) d; rhIL-11: 28 (12-80) d; blank group: 35 (18-86) d, P<0.05]. Platelet transfusions were also less in rhTPO group than in rhIL-11 and blank groups [20 (10-30) U, 30 (10-50) U, 35 (10-70) U, P<0.05]. The counts of megakaryocyte in rhTPO group, rhIL-11 group and blank group were 31.5 (0-200), 12 (0-142) and 11(0-187) (P<0.05), respectively. The difference between rhTPO group and rhIL-11 group was statistically significant (P<0.05), but no difference between rhIL-11 group and blank group (P>0.05). Multivariate analysis showed that rhTPO was an independent factor for platelet recovery [HR=4.01 (95%CI 1.81-9.97), P=0.010]. The rhTPO group had no obvious adverse events. Conclusion rhTPO can promote platelet recovery of SAA patients after allo-HSCT, reduce platelet transfusion with safety.

OBJECTIVETo assess the accuracy of quantitative fluorescence PCR(QF-PCR) for the detection of fetal chromosomal aneuploidies and its values for prenatal diagnosis.

METHODSQF-PCR and chromosomal karyotyping were used to analyze 6066 amniotic fluid samples derived from 6034 pregnant women.

RESULTSBoth QF-PCR and karyotyping analysis have detected 135 cases of fetal aneuploidies involving chromosomes 21, 18, 13, X, and Y. The QF-PCR assay was also successful in 67 cases for which amniotic fluid culture has failed. Furthermore, it has identified maternal cell contamination in 7 cases. By determining the consistency of short tandem repeat (STR) sites, the QF-PCR assay has identified 22 dizygotic twins among 32 twins with double chorions and double amniotic sacs. In 12 cases, it has signaled numerical chromosomal aberration by critical or partial abnormal values for the fluorescence peak area ratio, which were verified by karyotyping analysis as mosaicisms of chromosome aneuploidies.

CONCLUSIONThe QF-PCR can provide an useful supplement for chromosomal karyotyping and has an important role in rapid prenatal diagnosis.

Adolescent ; Adult ; Aneuploidy ; Female ; Fluorescence ; Humans ; Karyotyping ; Microsatellite Repeats ; Middle Aged ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis ; methods ; Young Adult

Objective To evaluate the efficacy and safety of autologous hematopoietic stem cell transplantation (AHSCT) in patients with relapsed and refractory malignant lymphoma. Methods The clinical data of 10 patients (6 males and 4 females) with relapsed (4 cases) and refractory (6 cases) malignant lymphomas who received AHSCT in General Hospital of Jinan Military Command from August 2011 to June 2015 were analyzed retrospectively. The median age was 34 years (20 ˉ50 years); 5 cases of Hodgkin lymphoma, 5 cases of non-Hodgkin lymphoma. Before transplantation, all patients received several courses of radiotherapy or chemotherapy. High dose of cytoxan combined with G-CSF were used to mobilize peripheral hematopoietic stem cell, and preconditioning regimen included BEAM, CBV or TBI. Results The median mononuclear cell of 10 patients was 7.385 × 108/kg. Complete remission was achieved in 8 patients after transplantation, and 2 cases relapsed. Median follow-up time was 18 months (20ˉ50 months). The overall survival rate and disease-free survival rate both were 80%(8/10). All patients had nausea, vomiting, diarrhea, oral mucositis and other adverse reactions, which could be tolerated. Conclusion ASHCT is an effective and safe method for treatment of relapsed and refractory malignant lymphomas.

Objective: To investigate the effects of PRX-2 gene on phenotype changes in epidermal stem cells differentiating into sweat gland cells. Methods: Epidermal stem cells and sweat gland cells separated and cultured from healthy foreskin and adult full-thick skin respectively, were identified by immunofluorescence staining. Lentiviral vector-mediated overexpression and knockdown of PRX-2 gene in epidermal stem cells were performed respectively, with empty vector-mediated epidermal stem cells as a control group. Overexpression、blank control and knowdown group′s PRX-2 expressions in gene and protein levels were detected using RT-PCR and Western blot technology. The ESCs of each group were co-cultured with sweat gland cells through transwell plate, and the expressions of CEA and β1 integrin in epidermal stem cells were determined by flow cytometry before and after co-culturing. Results: Epidermal stem cells and sweat gland cells were in line with their respective specific antigens. Before co-cultured, epidermal stem cells highly expressed β1 integrin (98.69±0.67)%, hardly expressed CEA (6.20±3.15)%. After co-cultured, β1 integrin expression levels were showed as knockdown group (19.30±0.53)% blank control group (51.20±0.79)%> overexpress group (45.91±0.93)%. There had significant differences between those of each two groups. Conclusions PRX-2 gene can inhibit the phenotypic change of Epidermal Stem Cells differentiating into Sweat Gland Cells and improve the ability to maintain their own specific antigens.

Tissue-engineered skin plays an important role in clinical applications,and even the rapid development of science and technology promotes the research about it.Choosing an appropriate animal model for wound repair is the prerequisite for the objective evaluation of the object of study.In this paper,the research progress of animal models of wound repair was introduced from several aspects,such as selection of experimental animals,making of wound models,skin-related cells and materials,wound healing evaluation indexes,etc.,hoping to provide reference for later research work.

Anemia, Aplastic ; therapy ; Hematopoietic Stem Cell Transplantation ; Humans

Objective To determine the effect of platelet-derived growth factor-C (PDGF-C) on biological characters of human hair dermal papilla cells cultured in vitro.Methods The human dermal papilla cells(HDPCs) were isolated from human hair skin obtained from rhytidectomy procedure and then cultured in vitro.Cell counting and CCK-8 assay were used to detect the effects of PDGF-C (0,10,20,30 and 40ng/ml) on the proliferation of HDPCs at 0,1 d,2d,3d,4d,5d,6d.Under the optimal concentration,flow cytometry was used to detect cell phases.Transwell assay and cell scratch test were performed to detect cell migration.Alkaline Phosphatase Activity Assay kit was used to detect the inductive activity of HDPCs.Results PDGF-C significantly induced the proliferation of HDPCs.PDGF-C of 30ng/mL promoted the HDPCs proliferation at a summit and increased the percentage of the cells arrested at S phase (P ＜ 0.05).PDGF-C also increased the migration populations of cultured HDPCs.The cell number in lower side of the transwell insert membrane of 30ng/ml PDGF-C treated group was (361.3 ± 24.95)while the control was (246.8 ± 7.525),showing significant difference (P ＜ 0.05).The alkaline phosphate activity of cultured HDPCs was increased comparing to the control group,the difference was significant (P ＜ 0.05).Conclusions PDGF-C can promote the proliferation,migration and inductive activity of cultured human dermal papilla cells,which might be beneficial to promote the cultivation of human dermal papilla cells in vitro.