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 value of pulse oxygen perfusion index(PI)and blood lactic acid(BLA)concentration in early prediction of retinopathy of prematurity(ROP).Methods A retrospective case-control study was conducted on 128 preterm infants who met the inclusion criteria and were admitted to the neonatal intensive care unit of our hospital from September 2018 to December 2022.Among them,46 patients with ROP were in the ROP group,and 82 pa-tients without ROP were in the non-ROP group.Basic data of these preterm infants were recorded after admission.PI val-ues were continuously monitored with the Masimo Radical-7(USA)SpO2 blood oxygen saturation detector,and BLA con-centrations were detected with the ABL90FLEX blood gas analyzer.The receiver operating characteristic(ROC)curve and area under curve(AUC)were used to evaluate the value of PI and BLA concentration in early prediction of ROP.Results There were no significant differences in gestational age,birth weight,sex,and delivery mode between the two groups(all P＞0.05).The PI values after birth were significantly different between the two groups(Fgroup=15.393,Pgroup＜0.001).The PI values of preterm infants in the ROP group decreased significantly at 1 h,12 h and 24 h after birth and slightly at 48 h to 96 h after birth compared with the non-ROP group.The PI values of preterm infants in the two groups sta-bilized at 96 h after birth.The PI values of preterm infants in the ROP group were lower than those in the non-ROP group at all time points within 96 h after birth(all P＜0.05).The PI values showed interaction effects between the two groups at different time points(Finteraction=5.061,Pinteraction＜0.001).There was a significant difference in BLA concentration between the two groups after birth(Fgroup=91.158,Pgroup＜0.001).In the ROP group,the BLA concentration increased significantly at 1 h after birth and slightly at 12 h and 24 h after birth compared with the non-ROP group.The BLA concentration in the ROP group was higher than that in the non-ROP group at all time points after birth(all P＜0.05).The BLA concentration showed no interaction effects between the two groups at different time points(Finteraction=0.567,Pinteraction＞0.05).The AUC of PI values at 1 h,12 h and 24 h after birth and BLA concentration at 1 h after birth for predicting ROP was 0.77,0.82,0.83,and 0.82,respectively.The AUC of combined PI values at 1 h,12 h and 24 h after birth and BLA concentration at 1 h after birth for predicting ROP was 0.94,higher than the predictive value of a single indicator.Conclusion PI and BLA concentration have good clinical value for early prediction of ROP.

Objective:To study the trinucleotide repeats of GCN (GCA, GCT, GCC, GCG) encoding Alanine in exon 3 of the PHOX2B gene among healthy individuals from southwest China and two patients with Congenital central hypoventilation syndrome (CCHS). Methods:The number and sequence of the GCN repeats of the PHOX2B gene were analyzed by capillary electrophoresis, Sanger sequencing and cloning sequencing of 518 healthy individuals and two newborns with CCHS, respectively. Results:Among the 1036 alleles of the 518 healthy individuals, five alleles were identified, including (GCN) 7, (GCN) 13, (GCN) 14, (GCN) 15 and (GCN) 20. The frequency of the (GCN) 20 allele was the highest (94.79%). And five genotypes were identified, which included (GCN) 7/(GCN) 20, (GCN) 13/(GCN) 20, (GCN) 14/(GCN) 20, (GCN) 15/(GCN) 20, (GCN) 20/(GCN) 20. The homozygous genotypes were all (GCN) 20/(GCN) 20, and the carrier rate was 89.58%. Four GCN sequences of the (GCN) 20 homozygous genotypes were identified among the 464 healthy individuals. The GCN repeat numbers in the exon 3 of the PHOX2B gene showed no significant difference between the expected and observed values, and had fulfilled the, Hardy-Weinberg equilibrium. The genotypes of the two CCHS patients were (GCN) 20/(GCN) 25 and (GCN) 20/(GCN) 30, respectively. Conclusion:It is important to determine the GCN repeats and genotypic data of the exon 3 of the PHOX2B gene among the healthy individuals. The number of GCN repeats in 518 healthy individuals was all below 20. The selection of appropriate methods can accurately detect the polyalanine repeat mutations (PARMs) of the PHOX2B gene, which is conducive to the early diagnosis, intervention and treatment of CCHS.

Objective:To analyze the difference of clinical characteristics and outcomes of infants with moderate and severe pediatric acute respiratory distress syndrome(PARDS)diagnosed according to baseline oxygenation index(OI) in pediatric intensive care unit(PICU).Methods:Second analysis of the data collected from the "Efficacy of pulmonary surfactant (PS) in the treatment of children with moderate and severe ARDS" program.Retrospectively compare of the differences in clinical data such as general condition, underlying diseases, OI, mechanical ventilation, PS administration and outcomes among infants with moderate and severe PARDS divided by baseline OI who admitted to PICUs at 14 participating tertiary hospitals from 2016 to December 2021.Results:Among the 101 cases, 55 cases (54.5%) were moderate and 46 cases (45.5%) were severe PARDS.The proportion of male in the severe group (50.0% vs.72.7%, P=0.019) and the pediatric critical illness score(PCIS)[72 (68, 78) vs.76 (70, 80), P=0.019] were significantly lower than those in the moderate group, while there was no significant difference regarding age, body weight, etiology of PARDS and underlying diseases.The utilization rate of high-frequency ventilator in the severe group was significantly higher than that in the moderate group (34.8% vs.10.9%, P=0.004), but there was no significant difference in PS use, fluid load and pulmonary complications.The 24 h OI improvement (0.26±0.33 vs.0.04±0.34, P=0.001) and the 72 h OI improvement[0.34 (-0.04, 0.62) vs.0.15 (-0.14, 0.42), P=0.029)]in the severe group were significantly better than those in the moderate group, but there was no significant difference regarding mortality, length of hospital stay and intubation duration after diagnosis of PARDS between the two groups. Conclusion:In moderate and severe(divided by baseline OI) PARDS infants with invasive mechanical ventilation, children in severe group have better oxygenation improvement in the early stage after PARDS identified and are more likely to receive high frequency ventilation compared to those in moderate group.Baseline OI can not sensitively distinguish the outcomes and is not an ideal index for PARDS grading of this kind of patient.

Objective:To investigate the effectiveness and safety of the allogeneic corneal stromal flap in reducing the exposure of the drainage catheter in the implantation of Ahmed glaucoma valve.Methods:Fifty-three patients(61 eyes) with refractory glaucoma treated by Ahmed glaucoma valve implantation from January 2017 to April 2019 were retrospectively analysed. The patients were divided into sclera tunnel group(routine group) and sclera tunnel with allogeneic corneal stroma flap covering(improved group). The cumulative success rate of surgery, preoperative intraocular pressure(IOP), IOP at the last outpatient follow-up, the best-corrected visual acuity(BCVA), BCVA at the last outpatient follow-up, drainage catheter exposure and other surgical complications were collected from both groups at 6, 12 and 24 months after surgery. The data between the 2 groups were statistically analyzed, P<0.05 was statistically significant. Results:The cumulative success rates of 6, 12 and 24 months after operation were 89.7%, 86.2% and 69.0% in the routine group, and 90.6%, 90.6% and 71.8% in the improved group, respectively. There was no significant differences between the 2 groups( P>0.05). The IOP and BCVA at the final follow-up were significantly improved in both groups, with a statistical significant difference( P<0.05). In the conventional group, the exposure of drainage catheter occurred in 4 eyes(13.8%). Further surgeries were carried out to cover the exposed drainage catheters with allogeneic corneal stroma flaps and amniotic membrane and all had good recovery. There was no drainage catheter exposure in the improved group. The difference between the 2 groups was statistically significant( χ2=4.724, P=0.030). There was no significant difference in other surgical complications between the 2 groups( χ2=0.160, P=0.689). No intraoperative or postoperative complications regarding the corneal stromal flap were observed. Conclusion:Implantation of Ahmed glaucoma valve with allogeneic corneal stromal flap sealing can effectively reduce the exposure of the drainage catheter. It is a safe method.

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 explore the role of lung dendritic cells (DCs) and Th17/regulatory T cells (Treg) pathway in the pathogenesis of chronic obstructive pulmonary disease(COPD).Methods COPD patients who received lobectomy from Sep.2015 to Mar.2016 in our hospital were enrolled and classified into non-smoking non-COPD group,smoking without COPD group and COPD group.The expression of CD80,chemokine recepter-6 (CCR6),interleukin-17A (IL-17A) and fork-head transcription factor P3 (FoxP3) were detected by immunohistochemistry (IHC) in lung tissue.Mature DCs (mDCs),immature DCs (imDCs),Th17 cells and Treg cells in lung tissue were detected by flow cytometry (FCM) and the correlation between Th17/ Treg cells with lung function was analyzed.Results (1) The expression of CD80 and FoxP3 in COPD group was decreased,while the expression of CCR6 and IL-17A was increased (P＜0.05).(2) The percentage of mDCs and Treg in lung tissue of COPD group was significantly decreased.In contrast,the proportion of imDCs and Th17 cells in COPD group was significantly increased (P＜0.05).(3) The imbalance of Th17/Treg ratio in lung tissue was seen in patients with COPD,suggesting the potential mechanism of Th17 cell-mediated proinflammatory response.(4) The percentage of Th17 cells and Th17/Treg ratio in COPD patients was negatively correlated with forced expiratory volume in the first second (FEV1) as a percentage of predicted value (FEV1％ pred),forced vital capacity(FVC) as a percentage of predicted value (FVC％ pred),FEV1/FVC.On the other hand,the percentage of Treg cells was positively correlated with FEV1％ pred,FVC％ pred,FEV1/FVC.Conclusions The data in this study demonstrate the maturation disorder of dendritic cells in lung tissue of COPD patients.The imbalance of Th17/Treg ratio suggests that Th17 cell-mediated proinflammatory response may be involved in the pathogenesis of COPD.

Objective To observe the efficacy and toxicity of thermotherapy plus chemotherapy in treatment of retroperitoneal lymph node metastasis after radical gastrectomy and sequential radiochemotherapy. Methods Sixty patients with retroperitoneal lymph node metastasis after radical gastrectomy and sequential radiochemotherapy were randomly divided into of microwave hyperthermia combined with chemotherapy group (experimental group, 30 cases) and chemotherapy group (control group, 30 cases) by using random number table method. The control group: oxaliplatin 130 mg/m2, intravenous drip (2 h), d1; S-1: 80 mg·m-2·d-1, P.O 2 times/d (after breakfast and after dinner), d1-14. 3 weeks was 1 cycle, a total of 4 cycles. The experimental group: chemotherapy on the basis of control group combined with microwave hyperthermia, d1, 8. the efficacy and toxicity of two groups were evaluated. Results The efficacy rate of the experimental group was higher than that of the control group, and the difference was statistically significant [66.7 % (20/30) vs. 33.3 %(10/30), P< 0.05]. The improvement of Karnofsky score in the experimental group was better than that in the control group, and the difference was statistically significant [73.3%(22/30) vs. 23.3%(7/30), P<0.05]. The improvement of pain score in the experimental group was better than that in the control group, and the difference was statistically significant [75.0 % (15/20) vs. 17.6 % (3/17), P< 0.05]. There was no significant difference in gastrointestinal reactions of two groups [30.0%(9/30) vs. 26.7%(8/30), P>0.05]. There was no significant difference in bone marrow suppression of two groups [33.3%(10/30) vs. 30.0 % (9/30), P> 0.05]. Conclusion Microwave thermotherapy plus chemotherapy has a good efficacy for retroperitoneal lymph node metastasis after radical gastrectomy and sequential radiochemotherapy, and the patients can well tolerated, it is worthy of clinical promotion.

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 construct a recombinant lentivirus containing human beta defensins -3 ( hBD3 ) , connective tissue growth factor gene (CTGF) and enhanced green fluorescent protein (EGFP), and to detect its translation in rabbit bone marrow mesenchymal stem cells (BMSC).Methods The lentivirus containing hBD3, CTGF and EGFP genes was constructed in vitro.The titer of lentivirus was tested with end-paint dilution assay .Rabbit BMSCs were transfected with recombinant virus.The best value of multiplicity of infection (MOI) was tested.The expression condition, transfection efficacy and genetic stability of the target genes were evaluated by using fluorescence microscopy and flow cytometry . Western blotting was used to detect the expression of the target protein .Results Recombinant lentivirus vectors: Lenti-CTGF-hBD3-EGFP, Lenti-hBD3-EGFP, and Lenti-EGFP, were successfully obtained . The titer of the recombinant lentiviruses was 3.21 ×108, 5.80 ×108, and 1.16 ×109, respectively.The best MOI value to transfect BMSCs was 150. The transfection efficacy of these lentivirus vectors was high , reaching 79.72%as assessed by flow cytometry , and it could be stably inherited .Western blotting displayed that target protein expression was successful .Conclusion The construction of recombinant lentiviruses carrying hBD3 and CTGF genes is successful and can be effectively transfected into BMSCs .