BACKGROUND: This study was undertaken to determine the effect of mesenchymal stem cells (MSCs) engraftment on vascular endothelial cell growth factor (VEGF) in lung tissue, plasma and extravascular lung water at early stage of smoke inhalation injury. METHODS: A rabbit smoke inhalation injury model was established using a home-made smoke inhalation injury generator, and rabbits were divided into two groups randomly: a control group (S group, n=32) and a MSCs treatment group (M group,n=32). 10 ml PBS was injected via the ear marginal vein immediately at injury into the S group. Third generation MSCs with a concentration of 1×107/10 ml PBS were injected via the ear marginal vein immediately at injury into the M group. VEGF in peripheral blood and lung tissue were measured at 0 (baseline), 2, 4 and 6 hours after injection respectively and analyzed. The right lungs of rabbits were taken to measure lung water mass fraction. RESULTS: In the lung tissue, VEGF decreased gradually in the S group (P<0.05) and signifi cantly decreased in the M group (P<0.05), but it increased more signifi cantly than the values at the corresponding time points (P<0.05). In peripheral blood, VEGF increased gradually in the S group (P<0.05) and markedly increased in the M group (P<0.05), but it decreased more signifi cantly than the values at corresponding time points (P<0.05). CONCLUSION: MSCs engraftment to smoke inhalation injury could increase VEGF in lung tissue, decrease VEGF in plasma and reduce extravascular lung water, indicating its protective effect on smoke inhalation injury.

China ; Heart Transplantation ; methods ; Humans

OBJECTIVETo study the effect of bone marrow-derived mesenchymal stem cells (MSC) transplantation on the major inflammatory cytokines content in peripheral blood, lung water mass fraction, and lung tissue injury in rabbits with smoke inhalation injury.

METHODSSixteen adult New Zealand big ear rabbits were subjected to smoke inhalation injury and then were divided into pure injury group (PI, n = 8) and MSC transplantation group (MT, n = 8) according to the random number table. Via ear marginal vein, rabbits in PI group were injected with 10 mL phosphate buffered saline (PBS); rabbits in MT group were injected with 10 mL PBS containing the third passage MSC (1 x 10⁷ cell) isolated from marrow of healthy young New Zealand big ear rabbit. Another 8 rabbits were enrolled as normal control group (NC). Rabbits in NC group were injected with 10 mL PBS via ear marginal vein without smoke inhalation injury. Blood was harvested from rabbits in PI and MT groups at post injury hour (PIH) 2, 4, 6. Contents of TNF-α, IL-1β, IL-6, and IL-10 in serum were determined with ELISA. At PIH 24, left lung was harvested for morphology and histopathology observation; the right lung tissue was obtained to measure and calculate lung water mass fraction. Blood and lung tissue of rabbits in NC group were harvested and determined in the same way. Data were processed with t test.

RESULTS(1) Serum contents of TNF-α in PI and MT groups at each time point were obviously higher than that in NC group (t = 2.43 - 9.57, P < 0.05 or P < 0.01). Serum contents of IL-1β and IL-6 in PI group at each time point were obviously higher than those in NC group (t = 8.49 - 19.80, P values all below 0.01). Serum content of IL-1β in MT group at each time point was close to that in NC group (t = 0.11 - 0.92, P values all above 0.05). Serum content of IL-6 in MT group at PIH 2 was close to that in NC group (t = 2.12, P > 0.05), but that of MT group increased significantly at PIH 4 and 6 (t = 2.83, P values all below 0.05). Serum contents of TNF-α, IL-1β, and IL-6 in MT group at each time point were obviously lower than those in PI group (t = 2.35 - 12.45, P < 0.05 or P < 0.01). (2) Serum content of IL-10 in MT group at PIH 2, 4, and 6 was respectively (13.0 ± 3.6), (11.6 ± 8.5), (15.2 ± 4.4) pg/mL, and they were higher than those in PI group [(5.5 ± 3.4), (5.0 ± 1.7), (7.9 ± 3.5) pg/mL, with t value respectively 4.28, 2.15, 3.67, P values all below 0.01]. Serum contents of IL-10 in PI and MT groups were obviously higher than that in NC group (t = 2.46-8.14, P < 0.05 or P < 0.01). (3) Lung tissue injury in MT group was alleviated markedly as compared with that in PI group. (4) At PIH 24, lung water mass fraction in MT group was (69 ± 7)%, which was obviously lower than that in PI group [(87 ± 6)%, t = 5.49, P < 0.01]. Compared with that in NC group [(48 ± 3)%], lung water mass fraction in PI and MT groups were increased obviously (with t value respectively 16.93 and 7.22, P values all below 0.01).

CONCLUSIONSMSC transplantation can decrease pro-inflammatory cytokines, increase anti-inflammatory cytokines, decrease lung water mass fraction, ameliorate systemic inflammatory response, and protect lung tissue in rabbits with smoke inhalation injury.

Animals ; Bone Marrow Cells ; cytology ; Inflammation ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Lung Injury ; blood ; surgery ; Mesenchymal Stem Cell Transplantation ; Rabbits ; Smoke Inhalation Injury ; blood ; surgery ; Tumor Necrosis Factor-alpha ; blood

Objective To investigate the characteristics of infecting pathogens,their changes and antimicrobial susceptibilities on CAPD related peritonitis in our peritoneal dialysis(PD) center in the past 15 years.Methods Two hundred and six CAPD related peritonitis episodes in 145 patients from 2000 to 2005 were analyzed and compared with 109 episodes from 1991 to 2000.The causative pathogens,their antimicrobial susceptibilities and outcomes on CAPD related peritonitis from the two periods were retrospectively reviewed and compared.Results Culture negative rate decreased from 60.6% in 1990 s to 47.6% in the last five years (P=0.031 ).Among culture positive peritonitis episodes,the incidence of gram positive bacteria (GPB) peritonitis increased from 25.6% to 39.8% (P=0.059).This was mainly due to a significant increase in coagulase-neagative staphylococcus peritonitis,which significantly increased from 4.7% to 26.9% (P=0.01).Gram negative bacteria (GNB) peritonitis decreased slightly (44.2% vs 34.3%,P=0.322).The incidence of Klebsiella pneumoniae peritonitis significantly decreased (14.0% vs 3.7%,P=0.023),while Pseudomonas aeruginosa and Escherichis coli peritonitis rates slightly increased (4.7% vs 9.3%,P = 0.338;7% vs 18.7%,P=0.072).The decrease of fungal peritonitis rate was not significant (30.2% vs 17.6%,P= 0.123).The comparison of clinical outcomes showed an improvement of total recovery rate from 68.8% in 1990 s to 73.9% for 2000-2005 (P=0.09).The catheter removal rate decreased from 19.2% to 14.3% (P=0.238),and the mortality from 10.1% to 5.4% (P=0.118).In both periods,fungal peritonitis had the poorest results,which all the patients either withdrew from PD or died.Conclusions Compared with that in 1990 s,the culture positive rate for CAPD related peritonitis in 2000-2005 has been greatly improved.Coagulase-negative staphylococcus is the most common causative pathogen.The mortality and catheter removal rate have been markedly reduced in the last five years.Fungal peritonitis is the most important reason for patients' dropout.

Objective To investigate the efficacy and safety of Urinary kallidinogenase(UK)combined with butylphthalide in patients with acute cardioembolic stroke(ACS).Methods A retrospective collection was performed for ACS patients diagnosed and treated in Hangzhou First People's Hospital from May 2022 to April 2023.According to the treatment protocol,ACS patients were divided into UK group and Butylphthalide+UK group.The clinical efficacy,neurological function,serum indexes(Hcy,NT-proBNP and VEGF)and prognosis of the two groups were compared after 2 weeks of treatment.Results A total of 86 ACS patients were included in the study,including 43 in the UK group and 43 in the Butylphthalide+UK group.After treatment,the total effective rate of treatment in the Butylphthalide+UK group was significantly higher than that in the UK group(P<0.05),and there was no significant difference in the incidence of adverse reactions between the two groups(P>0.05).In addition,the expression levels of Hcy and NT-proBNP in ACS patients in the Butylphthalide+UK group were significantly lower than those in the UK group(P<0.05),while the expression levels of VEGF were significantly higher than those in the UK group(P<0.05).The NIHSS score and mRS score of ACS patients in the Butylphthalide+UK group were significantly lower than those in the UK group(P<0.05).The rate of collateral circulation establishment in the Butylphthalide+UK group was significantly higher than that in the UK group(P<0.05).Conclusion Butaphthalide combined with UK has significant efficacy and high safety in ACS patients,which may be achieved by regulating the expression levels of serum Hcy,NT-proBNP and VEGF,thereby improving neurological function and promoting the establishment of collateral circulation.

Objective To assess the diagnostic accuracy of 64-slice computed tomography coronary angiography (64-SCTCA) in individuals with suspected coronary artery disease (CAD). Methods The study enrolled 285 individuals undergoing 64-SCTCA with calcium scoring and thereafter invasive coronary angiography (CAG) within 4 weeks for suspected CAD. Pretest probability of having obstructive CAD was determined using the Duke clinical score, which was estimated by type of chest discomfort, age, gender,and traditional risk factors and stratified into 3 levels of probability: low ( ≤ 30%, n = 80), intermediate (31% to 70%, n =92), and high ( ≥71%, n = 113). CAD was defined as the presence of at least one vessel of ≥50% coronary stenosis on CAG. Results The patient-based diagnostic accuracy of 64-SCTCA for detecting CAD according to CAG revealed a sensitivity of 81.2%, a specificity of 93.3%, a positive predictive value of 68.0% and negative predictive value of 96.6%. The CAD prevalence in the low,intermediate and high risk groups according to Duke probability was 46.3%, 72.8% and 82.3%,respectively. The sensitivity and positive predictive value were lower in the low probability group than those in the intermediate and high probability groups. For those with coronary artery Agatston calcium score ＞400, the diagnostic accuracy was linked with a higher sensitivity but lower specificity. The diagnostic value of 64-SCTCA for proximal and mid-segment of coronary artery was superior to that for distal segment. Conclusions 64-SCTCA is mainly indicated in individuals with an intermediate probability of having CAD. The diagnostic value of 64-SCTCA could be affected by coronary artery calcium, lesion location and vessel diameter.

AIM To investigate the flavor technology for Jiannaoning Mixture based on electronic tongue combined with preference index.METHODS Artificial sensory evaluation method was employed to screen the type of flavor agents,electronic tongue was applied to screening the consumption of flavor agents,and double-bottle preference test was adopted in the analysis of effects of rat gender and body weight on taste sensitivity,after which acceleration test was conducted.RESULTS Sucrose,stevioside and xylitol were taken as flavor agents,whose optimal consumptions were 15%,0.08%and 15%,respectively.Male rats demonstrated stable taste perception with the body weights of 220-280 g,and 0.08%stevioside displayed the best flavoring effect.After placed under accelerated condition for 3 months,the preparation exhibited stable appearance,pH value and contents of lobetyolin,salvianolic acid B,schizandrin,ruscogenin.CONCLUSION This stable and reliable method can provide reference for improving the terrible tastes of Jiannaoning Mixture and other oral liquid preparations of traditional Chinese medicines.

OBJECTIVETo observe the homing and differentiation of marrow-derived mesenchymal stem cells (MSC) transplanted intravenously in smoke inhalation injured rabbits.

METHODSThirty-two New Zealand big ear rabbits were divided into normal control group (NC), inhalation injury group (II), normal control + MSC treatment group (NM), and MSC treatment group (MT) according to the random number table, with 8 rabbits in each group. Rabbits in NC group were injected with 10 mL phosphate buffered saline (PBS) via ear marginal vein. Rabbits in NM group were injected with 10 mL PBS containing the third generation MSC labeled by BrdU (1 × 10(7) per 10 mL PBS) via ear marginal vein. Severe smoke inhalation injury model was reproduced in the other two groups, among them rabbits in II group were treated as rabbits in NC group, rabbits in MT group treated as rabbits in NM group. On the 7th and 28th day post treatment (PTD), lung tissue and trachea tissue were harvested from four groups for observation on injury with HE staining. Homing of MSC in injured tissue was observed with immunohistochemistry staining. The differentiation of MSC into functional cells was observed with immunohistochemical double staining of combining nuclear marker BrdU with lung (trachea) membrane-specific marker aquaporin-5 (AQP-5), alkaline phosphatase (AKP), CD34, and cytokeratin respectively.

RESULTS(1) MSC homing in lung and trachea tissue was observed in MT group on PTD 7, which was not observed in NM group. (2) AQP-5, AKP, and CD34 positive MSC were observed in lung tissue in MT group on PTD 28, while cytokeratin positive MSC was not observed in trachea tissue. No positively marked MSC was observed in NM group. (3) Injury in lung and trachea was less severe in MT group than in II group; and the proliferation of fibroblasts was less in MT group.

CONCLUSIONSIntravenous injection of MSC to rabbits with smoke inhalation injury can migrate to lung and trachea tissue at obviously inflammatory site, and differentiate into alveolar epithelial cells typeI and II, and pulmonary vascular endothelial cells, which may participate in the process of tissue repair in smoke inhalation injury.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Endothelial Cells ; cytology ; Epithelial Cells ; cytology ; Lung ; cytology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Pulmonary Alveoli ; cytology ; Rabbits ; Smoke Inhalation Injury ; pathology ; Trachea ; cytology

OBJECTIVETo increase the success rate of prenatal diagnosis for classical phenylketonuria(PKU).

METHODSThree new short tandem repeat (STR) markers (PAH26, PAH32 and PAH9) within and surrounding phenylalanine hydroxylase(PAH) gene were selected for amplified fragment length polymorphism. The allele frequencies and polymorphism information contests (PIC) were determined in Chinese population.

RESULTSThe PIC of these three new STR markers was 0.518 (PAH26), 0.413 (PAH32) and 0.362 (PAH9) respectively. There was linkage disequilibrium between PAH9 marker and PAH-STR marker (TCTA)n in the intron 3 of PAH gene. The linkage phase of the mutant genes and the markers was established using the combination of PAH-STR, PAH26 and PAH32 in 95% families. Prenatal diagnosis was performed successfully with these markers in four cases.

CONCLUSIONBy selecting or combining the three STR markers, the mutant genes could be distinguished from the normal allele in up to 95% of families with classical PKU.

Alleles ; Female ; Gene Frequency ; Genetic Linkage ; genetics ; Humans ; Linkage Disequilibrium ; Male ; Microsatellite Repeats ; genetics ; Mutation ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; diagnosis ; genetics ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis ; methods

OBJECTIVETo study the effects of high frequency oscillatory ventilation (HFOV) combined with incremental positive end-expiratory pressure (IP) on respiratory and circulatory functions, and lung histopathology of dogs with smoke inhalation injury.

METHODSAfter being treated with conventional mechanical ventilation, 12 dogs were inflicted with severe smoke inhalation injury and divided into group HFOV and group HFOV+IP according to the random number table, with 6 dogs in each group. Then they received corresponding ventilation for 8 hours respectively. Blood gas analysis results (pH value, PaO2 and PaCO2 levels) and hemodynamic parameters [heart rate, mean arterial pressure (MAP), pulmonary arterial pressure (PAP), central venous pressure (CVP), cardiac output (CO)] were recorded before injury, immediately after injury, and at post ventilation hour (PVH) 2, 4, 6, and 8. The dogs of two groups were sacrificed at PVH 8. A healthy dog without any treatment and a dog with smoke inhalation injury but no subsequent treatment were sacrificed in addition. Lung tissues of all dogs were obtained for histopathological observation. Lung injury score examination was conducted in both groups. Data were processed with rank sum test, analysis of variance of repeated measurement, and LSD- t test.

RESULTS(1) The PaO2 levels in both groups were significantly decreased immediately after injury, compared with those before injury (with t values respectively 4.960, 5.310, P values all below 0.01). The PaO2 levels in both groups from PVH 2 to PVH 8 were significantly increased, compared with those observed immediately after injury (with t values from 4.930 to 6.050, P values all below 0.01). At PVH 2, 4, and 8, PaO2 levels in group HFOV+IP were significantly higher than those in group HFOV (with t values from 3.775 to 5.774, P values all below 0.01); no statistically significant differences were observed in pH value and PaCO2 level at each time point between two groups (with t values from 0.002 to 0.997, P values all above 0.05). (2) There were no statistically significant differences in MAP, PAP, and CVP within two groups at each time point (with F values from 1.316 to 4.959, P values all above 0.05). In group HFOV, heart rate from PVH 2 to PVH 8 was significantly lower than that observed immediately after injury (with t values from 3.780 to 8.970, P values all below 0.01). In group HFOV+IP, CO at PVH 4, 6, and 8 was significantly lower than that observed immediately after injury (with t values from 3.990 to 11.200, P values all below 0.01). There were no statistically significant differences in MAP, PAP, and CVP between two groups at the same time point (with t values from 0.089 to 2.123, P values all above 0.05). At PVH 4, 6, and 8, heart rate in group HFOV+IP was higher than that in group HFOV (with t values from 2.931 to 7.229, P < 0.05 or P < 0.01), while CO was lower (with t values from 4.297 to 11.206, P values all below 0.01). (3) Compared with those of the healthy dog, inflammatory cell infiltration and bleeding in the lung were observed in alveolar space in both group HFOV and group HFOV+IP, while the degree was less serious than that of the dog with smoke inhalation injury only. Compared with those of group HFOV, inflammatory cell infiltration in group HFOV+IP was less significant, the alveolar structure was relatively intact, and no thickening of alveolar walls was observed. The lung injury score in group HFOV [(3.27 ± 0.24) points] was higher than that of group HFOV+IP [(2.79 ± 0.31) points, t = 27, P < 0.05].

CONCLUSIONSHFOV combined with IP can improve gas exchange and alleviate pulmonary injury without any adverse effect on blood gas analysis or hemodynamic parameters. Therefore, it may be considered as an appropriate mode of ventilation for the treatment of smoke inhalation injury.

Animals ; Blood Gas Analysis ; Burns, Inhalation ; physiopathology ; therapy ; Disease Models, Animal ; Dogs ; Hemodynamics ; High-Frequency Ventilation ; Male ; Positive-Pressure Respiration ; Smoke ; adverse effects