[Determination of 14 volatile organic compounds in urine by headspace gas chromatography-mass spectrometry].

Objective: To establish a automatic headspace gas chromatography-mass spectrometry (HS/GC-MS) method for the determination of 14 volatile organic compounds in urine. Methods: In September 2022, 10 ml urine sample was taken into a 20 ml headspace bottle, balanced for 30 min at 65 ℃, and then detected by HS/GC-MS and quantified by external standard method. Results: The 14 volatile organic compounds showed good linearity at 0.2-8.0 µg/L and 0.1-4.0 µg/L, with correlation coefficients ranging from 0.9956-0.9999. The recoveries were 79.8%-113.1% with relative standard deviations 0.05%-0.27% when three different concentration levels were added. Detection limit was 0.03-0.05 µg/L. Conclusion: The method is simple and convenient, and the recovery and precision meet the requirements. It can be used for the determination of common volatile organic compounds in urine.

[Sinusitis, otitis media and diffuse bronchiectasis in both lungs].

Primary ciliary dyskinesia (PCD) is a rare autosomal recessive or X-linked biallelic mutations inherited disease, characterized by motile cilia dysfunction. Typical manifestations include bronchiectasis, secretory otitis media, sinusitis, situs inversus, and infertility. PCD often needs to be differentiated from cystic fibrosis (CF) because of similar clinical manifestations. In this paper, a juvenile female who presented with recurrent cough and expectoration with fever since early childhood, had a history of secretory otitis media and sinusitis, and had been considered for the diagnosis of CF. After the discovery of compound heterozygous mutations in PCD related pathogenic genes by gene sequencing, combined with the clinical manifestations and imaging characteristics, PCD was finally diagnosed.

Nanoscale extracellular vesicle-derived DNA is superior to circulating cell-free DNA for mutation detection in early-stage non-small-cell lung cancer.

Background: The comparison between relatively intact nanoscale extracellular vesicle-derived DNA (nEV-DNA) and fragmented circulating cell-free DNA (cfDNA) in mutation detection among patients with non-small-cell lung cancer (NSCLC) has not been carried out yet, and thus deserves investigation. Patients and methods: Both nEV-DNA and cfDNA was obtained from 377 NSCLC patients with known EGFR mutation status and 69 controls. The respective EGFRE19del/T790M/L858R mutation status was interrogated with amplification-refractory-mutation-system-based PCR assays (ARMS-PCR). Results: Neither nEV-DNA nor cfDNA levels show a strong correlation with tumor volumes. There is no correlation between cfDNA and nEV-DNA levels either. The detection sensitivity of nEV-DNA and cfDNA using ARMS-PCR in early-stage NSCLC was 25.7% and 14.2%, respectively, with 96.6% and 91.7% specificity, respectively. In late-stage NSCLC, both nEV-DNA and cfDNA show ∼80% sensitivity and over 95% specificity. Conclusions: nEV-DNA is superior to cfDNA for mutation detection in early-stage NSCLC using ARMS-PCR. However, the advantages vanish in late-stage NSCLC.

[The clinical characteristics of 17 cases of primary ciliary dyskinesia].

Objective: To review the clinical data of cases of primary ciliary dyskinesia (PCD), and to explore the clinical characteristics for the understanding of PCD. Methods: We retrospectively summarized 17 patients with PCD diagnosed in Peking Union Medical College Hospital from Jan 2009 to Dec 2014. There were 7 male and 10 female patients, with the age from 6 to 57 years at the time of diagnosis. The mean onset age of the disease was 11.7±2.1 years, and the mean age at diagnosis was 29.5±3.5 years. We analyzed their clinical symtoms, radiologic images, pulmonary function test and the electron microscopic findings for the clinical characteristics of PCD. Results: The most common onset symptoms were cough (15/17) or sputum (13/17) among our 17 patients with PCD. Only 5 patients had situs inversus in our group. Sixteen patients had bronchiectasis on chest CT scan. Nasal sinusitis was confirmed by nasal CT scan in 15 patients. The most common pathogens from sputum cultures included Pseudomonas aeruginosa, Haemophilus influenzea, Aspergillus fumigates and Candida (3/14 respectively). None of the patients had evidence of mycobacterial infection. Twelve patients underwent spirometry and obstructive pattern was the most common disorder (8/12). Diffusion impairment (5/10) and restrictive pattern dysfunction (3/10) were also present in our patients. Two patients had normal pulmonary function test results. Thirteen patients underwent bronchial mucosal ciliary electron microscopy and the most abnormalities were outer or inner dynein arms deficiency (8/13), and other abnormalities included microtubule arrangement disorder (3/13), reduction of the number of microtubules (3/13) and reduction of the number of dynein arms (2/13). Four of our patients had multiple dysfunctions on their ciliaries. Conclusions: PCD should be considered in bronchiectasis patients with disease onset at childhood even without situs inverse, especially those accompanied with nasal sinusitis or otitis media. Chest or paranasal sinus CT scan, and male sperm examination were helpful for the diagnosis. Mucosal ciliary electron microscopy was an efficient diagnostic method for PCD.

Influence of sirolimus-induced TGF-ß secretion on mouse Treg cell proliferation.

We examined the effects of co-culturing CD4+ CD25+ Treg cells with sirolimus or cyclosporin A on Treg cell proliferation and differentiation and on transforming growth factor-ß (TGF-ß) and Foxp3 expression. CD4+ CD25+ Treg cells were harvested from mononuclear cells of spleens of C57BL/6 mice using immunomagnetic beads and divided into control, sirolimus, and cyclosporine groups. Following a 96-h co-culture, Treg cells were assayed by flow cytometry. FoxP3 and TGF-ß mRNA levels and secretion were assayed by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Smad protein of the TGF-ß signaling pathway was assayed by western blot and its effect on CD4+ CD25+ FoxP3+ Treg cell proliferation was determined. Sirolimus-promoted differentiation and proliferation was examined using a TGF-ß neutralizing antibody. Sirolimus-treated CD4+ T cell TGF-ß secretion increased 2.5X over control levels (P < 0.01), but that of the cyclosporine group decreased marginally (P > 0.05). The CD4+ cell proportion decreased significantly (41.25 vs 69.22%, P < 0.01) and slightly (65.21 vs 69.22, P > 0.05) in the cyclosporine and sirolimus groups, respectively. T cell Foxp3 mRNA expression was significantly higher in the sirolimus-treated than in the cyclosporine (53.7 vs 40.2%, P < 0.05) and control groups (P < 0.01), but was significantly lower in the cyclosporine group than in controls (23.6 vs 40.2%, P < 0.01). Overall, sirolimus promoted CD4+ CD25+ Treg cell proliferation and growth in vitro, whereas cyclosporin A inhibited proliferation. Sirolimus might promote CD4+ CD25+ FoxP3+ regulatory T cell proliferation by inducing TGF-ß secretion in vivo.

Differential diagnostic value of 3.0T MR 3D-ASL technique for recurrence and pseudo-progression of high-grade glioma.

OBJECTIVE: This study aimed to explore the value of 3.0T magnetic resonance three-dimensional arterial spin labeling imaging (3D-ASL) technology in the differential diagnosis of recurrence and pseudo-progression of high-grade gliomas. PATIENTS AND METHODS: Fifty patients with high-grade glioma were selected as research objects. All 50 patients were examined by magnetic resonance imaging (MRI), and the lesions were found to be enlarged or abnormally enhanced. All the patients were examined using the 3.0T MR 3D-ASL technique. With targeted biopsy pathology as the gold standard, the diagnostic results of the 3.0T MR 3D-ASL technique were analyzed, and the cerebral blood flow (rCBFmax) ratio was compared between patients with recurrent glioma and patients with pseudo-progression [maximum blood flow value/contralateral mirror area (CBFmax/contralateral mirror area), CBFmax/contralateral white matter, CBFmax/contralateral gray matter]. RESULTS: Among 50 glioma patients, 31 (62.00%) were diagnosed with recurrence through pathological examination, and 19 (38.00%) were diagnosed with pseudo-progression. 30 patients with recurrence (60.00%) and 20 patients with pseudo-progression (40.00%) were diagnosed using 3.0T magnetic resonance 3D-ASL technology. The diagnostic accuracy of 3.0T magnetic resonance 3D-ASL technology was 96.77% (30/31) (p > 0.05). Using pathological results as the "gold standard", the relevant parameters of 3.0T magnetic resonance 3D-ASL technology under different pathological results were analyzed. The results showed that the CBFmax/contralateral mirror area, CBFmax/contralateral white matter, and CBFmax/contralateral gray matter ratios of advanced glioma recurrence patients were significantly higher than those of pseudo-progression (p < 0.05). CONCLUSIONS: The application of 3.0T MR 3D-ASL in high-grade glioma can effectively distinguish recurrence and pseudo-progression, with significant diagnostic value.

A comparative study assessing a new tool for occluding parenchymal blood flow during liver resection for hepatocellular carcinoma.

BACKGROUND: The aim of this study was to compare the efficacy of a new tool (the hepatic section vascular blocker, HSVB) with hepatic pedicle clamping and hemihepatic vascular exclusion to control bleeding during liver resection for cancer. METHODS: Clinical data on 117 patients who underwent liver resection from 2004 to 2009 were analysed retrospectively. Forty-two patients had liver resection using the HSVB (group A), in 35 patients hemihepatic vascular exclusion was used (group B), and in 40 patients hepatic pedicle clamping with a Pringle manoeuvre was used (group C). Blood loss, operative time, postoperative hepatic function and complications were compared. RESULTS: Mean blood loss and operative time in group A were significantly less than in groups B (p=0.026 and p<0.001, respectively) and C (p<0.001 and p<0.001). There were significant differences between groups A and C in total bilirubin (TB) and alanine transaminase (ALT) levels on postoperative days 3 and 7, and group A had better hepatic function (TB p=0.014 and p=0.009; ALT p<0.001 and p<0.001). The rate of postoperative ascites was significantly higher in group C compared with group A (p<0.001). In group C, 2 patients had liver failure, 1 had a gastro-intestinal haemorrhage and 1 died. CONCLUSIONS: Using the HSVB during liver resection effectively controlled bleeding, saved operative time and preserved hepatic function. It proved to be a safe and feasible technique.

[Influence of work engagement and self-efficacy of nurses on clinical practice ability in burn intensive care unit].

Objective: To analyze the influence of work engagement and self-efficacy of nurses on clinical practice ability in burn intensive care unit (BICU), and to explore its potential pathways of action. Methods: A cross-sectional survey was conducted. From May to October 2020, a total of 30 hospitals with BICU in China were selected by stratified sampling method. Among BICU nurses who met the inclusion criteria, their clinical practice ability, work engagement, and self-efficacy were evaluated by self-evaluation scale of oriented problem-solving behavior in nursing practice (OPSN), Utrecht work engagement scale (UWES), and general self-efficacy scale (GSES), respectively. The total scale scores of each index and the average item scores were recorded. The self-designed general data questionnaire was used to investigate the nurses' gender, age, marital status, education background, working years, professional title, and the economic region of the hospital that they belonged to. The total scale scores of the above-mentioned three evaluation indexes were compared after the classification of nurses according to general data, and the data were statistically analyzed with independent sample t test or one-way analysis of variance. Pearson correlation analysis was used to analyze the correlation between the total scale scores of the three evaluation indexes. Based on the total scale scores of the above-mentioned three evaluation indexes, a structural equation model was established, the mediation analysis of the relationship among the three evaluation indexes and the pathway analysis of the structural model were conducted, and the Bootstrap method was used to verify the pathways of action. Results: A total of 401 questionnaires were distributed, and 337 valid questionnaires were returned, with a valid return rate of 84.04%. The total scale scores of clinical practice ability, work engagement, and self-efficacy of 337 nurses were 98.2±11.7, 67.7±18.6, and 26.6±5.6, respectively, and the average item scores were 3.9±0.5, 4.5±1.2, and 2.7±0.6, respectively. Among the 337 nurses, the majority were female, aged 40 or below, married, and had a bachelor's degree with work experience of ≤10 years; both nurses with professional nurse title and nurses from the Southeast region accounted for about 50%. There were statistically significant differences in the total scale score of clinical practice ability among nurses with different ages, education backgrounds, working years, and professional titles (with F values of 3.26, 4.36, 3.12, and 2.80, respectively, P<0.05). There was statistically significant difference in the total scale score of work engagement among nurses with different working years (F=4.50, P<0.05). There were statistically significant differences in the total scale score of self-efficacy among nurses with different ages, working years, and professional titles (with F values of 4.91, 4.50, and 2.91, respectively, P<0.05). The total scale score of nurses' work engagement was significantly positively correlated with the total scale score of clinical practice ability and the total scale score of self-efficacy (with r values of 0.30 and 0.51, respectively, P<0.05). The total scale score of nurses' self-efficacy was significantly positively correlated with the total scale score of clinical practice ability (r=0.37, P<0.05). The model had good adaptability, and the intermediary model was established. Nurses' work engagement had a significantly positive effect on both self-efficacy and clinical practice ability (with ß values of 0.54 and 0.16, respectively, P<0.05), and nurses' self-efficacy had a significantly positive effect on clinical practice ability (ß=0.29, P<0.05). Work engagement had a direct effect on self-efficacy and clinical practice ability, and self-efficacy had a direct effect on clinical practice ability and played a mediating role between work engagement and clinical practice ability. Bootstrap validation showed that self-efficacy played a significantly mediating role in the influence of work engagement on clinical practice ability (with effect size of 0.16, with 95% confidence interval of 0.08-0.24, P<0.05), accounting for half of the total effect of work engagement on clinical practice ability (with effect size of 0.32). Conclusions: BICU nurses have an above-average level of clinical practice ability, a medium level of self-efficacy, and a high level of work engagement. Work engagement and self-efficacy are positively correlated with clinical practice ability. Work engagement can directly affect clinical practice ability or indirectly affect clinical practice ability through the mediating role of self-efficacy.

[Feasibility, safety and long-term efficacy of laparoscopic total gastrectomy combined with distal pancreaticosplenectomy for T4b gastric cancer].

Objective: To explore the feasibility, safety and long-term efficacy of laparoscopic total gastrectomy combined with distal pancreaticosplenectomy for the treatment of T4b gastric cancer. Methods: A retrospective cohort study was performed. Clinical data of consecutive patients with T4b gastric cancer invading pancreatic tail undergoing laparoscopic or open total gastrectomy combined with distal pancreaticosplenectomy from January 2010 to December 2014 were analyzed retrospectively. Enrollment criteria: (1) primary gastric cancer confirmed by pathology as T4b adenocarcinoma; (2) chest+abdominal+pelvic enhanced CT indicated cancer invading pancreatic tail without distant metastasis, and R0 resection was evaluated as feasible before operation; (3) physical status was ECOG score 0 to 2, and was tolerant to operation. Patients with peritoneal implant metastasis and tumor invasion of other organs during operation, or changes in surgical methods for other reasons were excluded. All the operations were performed by the same surgical team, which had the experiences of more than 100 cases of laparoscopic and 100 cases of open radical gastrectomy with D2 lymph node dissection. The choice of surgical procedure was discussed by the surgeon and the patient, and decided according to the patient's intension. Patients were divided into the laparoscopic group and open group according to the surgical method. Intraoperative and perioperative findings were compared between the two groups. The 3-year disease-free survival rate were analyzed with Kaplan-Meier survival curve and compared by using log-rank test. Results: A total of 37 consecutive patients were enrolled, including 21 in the laparoscopic group and 16 in the open group, and no one receiving laparoscopic procedure was converted to open surgery. The baseline data of two groups were comparable (all P>0.05). Compared with the open group, the laparoscopic group had significantly longer operation time [(264.0±35.1) minutes vs. (226.6±49.9) minutes, t=2.685, P=0.011], significantly less intraoperative blood loss [(65.7±37.4) ml vs. (182.2±94.6) ml, t=-4.658, P<0.001], significantly shorter time to postoperative flatus [(2.8±0.7) days vs. (4.1±0.7) days, t=-5.776, P<0.001] and significantly shorter postoperative hospital stay [(13.3±2.8) days vs. (16.6±4.3) days, t=-2.822, P=0.008]. Morbidity of postoperative complications, including anastomotic leakage, pancreatic fistula, abdominal abscess, intraperitoneal hemorrhage and duodenal stump leakage, in two groups was similar [19.0% (4/21) vs. 4/16, P=0.705]. There were no cases of anastomotic bleeding or stenosis. The 30-day postoperative mortality was 0 in the laparoscopic group and 1/16 in the open group, respectively (P=0.432). The 3-year disease-free survival rates were 38.1% and 37.5% in the laparoscopic and open group, respectively (P=0.751). Conclusion: Laparoscopic total gastrectomy combined with distal pancreaticosplenectomy performed by experienced surgeons for T4b gastric cancer is safe and effective.

The efficacy and safety of immune checkpoint inhibitors in non-small cell lung cancer patients of different age groups: a meta-analysis.

BACKGROUND: Age is closely related to the efficacy of treatment for non-small cell lung cancer (NSCLC) patients. Latest clinical trials have proved the better overall survival (OS) for the use of immune checkpoint inhibitors verse chemotherapy in NSCLC patients. However, we had no clear idea of the efficacy of them in elderly patients. So we conducted a meta-analysis to compare the efficacy of immune checkpoint inhibitors for NSCLC patients of different age groups and summarized overall treatment-related adverse events. MATERIALS AND METHODS: PubMed, EMBASE, Web of Science and the Cochrane Library were searched for all clinical trials in NSCLC until 30th of April 2019. Eligible studies included randomized controlled trials (RCTs) comparing immune checkpoint inhibitors with chemotherapy in NSCLC patients. The hazard ratio (HRs) and 95% confidence intervals (CIs) of OS, progression-free survival or adverse events (AEs) were used. RESULTS: A total of 4994 patients from 8 RCTs were included. Immune checkpoint inhibitors significantly prolonged the OS (HR, 0.73; 95% CI, 0.61-0.89) versus chemotherapy in NSCLC patients who were less than 65 years old. Also, they prolonged the OS (HR, 0.74; 95% CI, 0.59-0.93) in NSCLC patients who were more than 65 years old. However, there was no statistical significance of OS (HR, 0.87; 95% CI, 0.57-1.30) among NSCLC patients who were more than 75 years old. It also showed that the single use of immune checkpoint inhibitors had fewer all-grade AEs. CONCLUSION: Regardless of the NSCLC patients who were less or more than 65 years, immune checkpoint inhibitors could achieve better OS than chemotherapy. But there was no significant difference when NSCLC patients who were more than 75 years old. Older patient should be offered immune therapies if it is possible and the mechanism in old age treatment should be further studied.

OTX1 is a novel regulator of proliferation, migration, invasion and apoptosis in lung adenocarcinoma.

OBJECTIVE: Orthodenticle Homeobox 1 (OTX1) has been found to be closely related to the development of several human tumours. However, the function and underlying molecular mechanisms of OTX1 in non-small cell lung cancer (NSCLC) are unclear. This research was performed to investigate the effects of downregulating OTX1 gene expression on the proliferation, migration, invasion, cell cycle and apoptosis of human NSCLC cell lines. PATIENTS AND METHODS: Cultured NCI-H292 and XWLC cells were transfected with control small interfering RNA (siNC) or experimental siRNA (siOTX1). The mRNA levels were detected using a quantitative real-time PCR (RT-qPCR) assay. A Cell Counting Kit-8 (CCK-8) and a Real Time Cell Analyzer (RTCA) were used to determine cell activity. The RTCA and transwell chambers were used to assess cell migration and invasion. In addition, cell cycle progression and apoptosis were measured using flow cytometry, and the expression levels of key signalling pathway proteins were examined by Western blotting. RESULTS: The results revealed that compared with the control group, the experimental group exhibited significantly decreased cell activity (***p<0.001), significantly decreased migration and invasion abilities (***p<0.001), and cell cycle arrest in G2/M phase (*p<0.05). However, the number of apoptotic cells was higher in the experimental group than in the control group (*p<0.05). The Western blotting results were consistent with the functional experiment results. CONCLUSIONS: Silencing the OTX1 gene suppressed the proliferation, migration and invasion of NCI-H292 and XWLC cells, impeded the cell cycle transition from G2 to M phase, and accelerated apoptosis, revealing OTX1, a regulator of NSCLC, as a potential new therapeutic target.

MiR-155-5p inhibits the proliferation and migration of VSMCs and HUVECs in atherosclerosis by targeting AKT1.

OBJECTIVE: MiR-155-5p has various biological cellular functions in diverse pathology, including cardiovascular disease. Nevertheless, the role of miR-155-5p in atherosclerosis is still not well known. PATIENTS AND METHODS: The levels of miR-155-5p and AKT Serine/Threonine Kinase 1 (AKT1) in plasma samples from patients with atherosclerotic CAD were detected using quantitative Real-time PCR (qRT-PCR). Cell counting kit-8 (CCK-8) assay was used to analyze the proliferation of vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs) in vitro. The migration of VSMCs and HUVECs was detected using wound healing assay. The invasion of VSMCs and HUVECs using was determined using the transwell invasion assay. The expression of AKT1 was measured using immunofluorescence staining analysis. RESULTS: MiR-155-5p was down-regulated in patients with atherosclerotic CAD. Up-regulation of miR-155-5p inhibited the proliferation, migration and invasion of VSMCs and HUVECs. Bioinformatics analysis and luciferase reporter assay indicated that AKT1 was the direct target of miR-155-5p and miR-155-5p bound to the 3'-untranslated region (3'-UTR) of AKT1. The expression of AKT1 was reduced in cell that was transfected with miR-155-5p. Up-regulation of AKT1 rescued the suppressive effect of miR-155-5p on the growth, migration and invasion of VSMCs and HUVECs. Down-expression of AKT1 partially neutralized the impacts of miR-155-5p on the growth, invasion and migration of VSMCs and HUVECs. Finally, we found that AKT1 was over-regulated in plasma samples of patients with atherosclerotic CAD and its level was negative with the level of miR-155-5p. CONCLUSIONS: Our study demonstrates that miR-155-5p suppresses the proliferation, migration and invasion of VSMCs and HUVECs through regulating AKT1, which provides the new insights into the precise role of miR-155-5p in atherosclerosis.

The lncRNA HOXA11-AS promotes glioma cell growth and metastasis by targeting miR-130a-5p/HMGB2.

OBJECTIVE: Long noncoding RNAs (lncRNAs) serve as important regulators of diverse types of cancer, including glioma. Nevertheless, their precise roles in cancers remain sufficiently unexplored. PATIENTS AND METHODS: Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was used to determine the levels of HOMEOBOX A11 antisense RNA (HOXA11-AS) and miR-130a-5p in glioma tissues and cell lines. Short hairpin RNAs (shRNAs) targeting HOXA11-AS or pcDNA3.1 were transfected into cells via a vector encoding HOXA11-AS to decrease or increase the level of HOXA11-AS. Cell Counting Kit-8 (CCK-8), colony formation, wound healing, flow cytometry and transwell assays were applied to assess the role of HOXA11-AS in glioblastoma cell growth, apoptosis and aggressiveness. The expression of N-cadherin and E-cadherin was determined using immunofluorescence staining. The expression of high-mobility group protein B2 (HMGB2) was determined using Western blot analysis in vitro and immunohistochemistry (IHC) staining in vivo. The direct target of HOXA11-AS and miR-130a-5p was confirmed using the Luciferase reporter assay. Glioblastoma cells were subcutaneously implanted into nude mice to determine the role of HOXA11-AS in tumor growth in vivo. RESULTS: In the current study, we demonstrated that the lncRNA HOXA11-AS was overexpressed in glioma. The overexpression of HOXA11-AS was correlated with advanced stages of glioma and poor prognosis. Downregulating HOXA11-AS expression significantly suppressed the proliferation, migration and invasion of glioma cells and increased their apoptosis. The growth of glioma cells in vitro was also suppressed by the downregulation of HOXA11-AS. Finally, we revealed that HOXA11-AS exerted its oncogenic effects by binding to miR-130a-5p, thereby neutralizing the suppressive effect of miR-130a-5p on HMGB2. CONCLUSIONS: Our results demonstrate that HOXA11-AS regulates the growth and metastasis of glioma by targeting the miR-130a-5p-HMGB2 signaling axis.

[Effects of allogeneic bone marrow mesenchymal stem cells on polarization of peritoneal macrophages in rats with sepsis].

Objective: To explore the effects of allogeneic bone marrow mesenchymal stem cells (BMSCs) on polarization of peritoneal macrophages isolated from rats with sepsis induced by endotoxin/lipopolysaccharide (LPS). Methods: (1) BMSCs were isolated, cultured and purified from 5 SD rats with whole bone marrow adherent method. The third passage of cells were collected for morphologic observation, detection of expressions of stem cell surface markers CD29, CD44, CD45, and CD90 with flow cytometer, and identification of osteogenic and adipogenic differentiation. (2) Another 45 SD rats were divided into sham injury group (SI, n=5), LPS control group (LC, n=20), and BMSCs-treated group (BT, n=20) according to the random number table. Rats in groups LC and BT were injected with LPS (5 mg/kg) via tail vein to induce sepsis; rats in group SI were injected with the same amount of normal saline to simulate the damage. At post injury hour (PIH) 1, rats in group BT were given 1 mL BMSCs (2×10(6)/mL) via tail vein injection; rats in another two groups were injected with equal volume of phosphate buffer saline. Five rats in group SI at PIH 24 and in groups LC and BT at PIH 6, 12, 24, and 48 were sacrificed to harvest lung tissue for pathological observation with HE staining. In addition, rats in group SI at PIH 24 and in groups LC and BT at PIH 24 and 48 were simultaneously performed with intraperitoneal injection of low-glucose DMEM. Then peritoneal fluid was harvested to culture peritoneal macrophages. Flow cytometer was used to assess the positive expression of cell makers of macrophages including CD68 (making gate), CD11c, and CD206 in group SI at PIH 24 and in groups LC and BT at PIH 24 and 48. Data were processed with one-way analysis of variance and LSD test. Results: (1) The third passage of cells showed uniform fiber-like shape similar to fibroblasts. These cells showed positive expressions of CD29, CD44, CD90 and weak positive expression of CD45. They were able to differentiate into osteoblasts and adipocytes. These cells were identified as BMSCs. (2) At PIH 24, the structure of pulmonary alveoli of rats in group SI was clear and complete with no congestion or inflammatory cell infiltration. At PIH 6, the structure of pulmonary alveoli of rats in groups LC and BT was clear with a small amount of inflammatory cell infiltration, slight congestion and pulmonary interstitial thickening. At PIH 12, the inflammatory responses in lung tissue of rats in group LC were more severe than those in group BT with a large amount of inflammatory cell infiltration, serious congestion, and obvious pulmonary interstitial thickening. The pathological results of rats in group BT at PIH 12 was consistent with the results at PIH 6. At PIH 24, the pathological results of rats in groups LC and BT were similar to the results at PIH 12. At PIH 48, the structure of pulmonary alveoli tissue of rats in group LC was still severely disrupted, with a large number of inflammatory cell infiltration and congestion in lung tissue, but pulmonary interstitial thickening was slightly alleviated than before. The condition of rats in group BT nearly recovered to that in group SI. (3) At PIH 24, the positive expression rate of CD11c in peritoneal macrophages of rats in group LC [(83±10)%] was close to that in group BT [(87±7)%, P>0.05], and they were both significantly higher than the rate in group SI [(55±12)%, with P values below 0.01]. The positive expression rate of CD11c in peritoneal macrophages of rats in group LC [(59±11)%] at PIH 48 was close to that in group SI at PIH 24 (P>0.05), and they were both significantly higher than the rate in group BT [(20±11)%] at PIH 48 (with P values below 0.01). At PIH 24, the positive expression percentages of CD206 in peritoneal macrophages of rats were similar among the three groups (with P values above 0.05). The positive expression percentage of CD206 in peritoneal macrophages of rats in group SI at PIH 24 was close to that in group BT at PIH 48 (P>0.05), and they were both significantly lower than the percentage in group LC at PIH 48 (with P values below 0.01). Conclusions: BMSCs can reduce the pathological inflammatory responses in the lung of rats with sepsis and inhibit peritoneal macrophages from polarizing into M1 phenotype, whereas they can not promote macrophages to polarize into M2 phenotype.

Evidence for the existence of an unfolding intermediate state for aminoacylase during denaturation in guanidine solutions.

The equilibrium unfolding of pig kidney aminoacylase in guanidinium chloride (GdmCl) solutions was studied by following the fluorescence and circular dichroism (CD). At low concentrations of GdmCl, less than 1.0 M, the fluorescence intensity decreased with a slight red shift of the emission maximum (from 335 to 340 nm). An unfolding intermediate was observed in low concentrations of denaturant (between 1.2 and 1.6 M GdmCl). This intermediate was characterized by a decreased fluorescence emission intensity, a red-shifted emission maximum, and increased binding of the fluorescence probe 1-anilino-8-naphthalenesulfonate. No significant changes of the secondary structure were indicated by CD measurement. This conformation state is similar to a molten globule state which may exist in the pathway of protein folding. Further changes in the fluorescence properties occurred at higher concentrations of GdmCl, more than 1.6 M, with a decrease in emission intensity and a significant red shift of the emission maximum from 340 to 354 nm. In this stage, the secondary structure was completely broken. A study of apo-enzyme (Zn2+-free enzyme) produced similar results. However, comparison of the changes of the fluorescence emission spectra of native (Holo-) enzyme with Zn2+-free (Apo-) enzyme at low GdmCl concentrations showed that the structure of the Holo-enzyme was more stable than that of the Apo-enzyme.

Protective Effect of Antiapoptosis Potency of Prolonged Preservation by Desiccation Using High-Pressure Carbon Monoxide on Isolated Rabbit Hearts.

Heart transplantation has been widely accepted as a therapy for end-stage heart failure. Mitigation of ischemia-reperfusion injury by inhibiting the apoptotic process plays an important role in organ transplantation. Desiccation using high-pressure carbon monoxide (CO) is a new method of preserving donor hearts; however, its mechanism of antiapoptosis remains unclear. This study was intended to elucidate the efficacy and mechanism of preservation by desiccation for 18 hours using high-pressure CO on myocardial apoptosis. Rabbit heterotopic abdominal cardiac transplantation models were established. New Zealand rabbits were divided randomly into 3 groups: naive group (n = 16), HTK group (n = 16), and desiccation using high-pressure CO group (n = 16). The donor hearts of the naive group were transplanted immediately after being extracted. In the HTK group, the donor hearts were extracted and steeped in 4°C HTK cardioplegic solution for 18 hours and then transplanted; in the desiccation using high-pressure CO group, the donor hearts were extracted and exposed to a gas mixture (Po2 = 3200 hPa, Pco = 800 hPa) in the chamber before being preserved in a refrigerator at 4°C for 18 hours and then transplanted. Apoptotic cardiomyocytes were detected using TUNEL technique and histopathology was performed by hematoxylin-eosin staining. The expression of the ratio of Bcl-2/Bax and caspase-3 proteins was detected using the Western blot method. These findings suggest that compared with traditional HTK preservation, preservation by desiccation using high-pressure CO could alleviate rabbits' myocardial histopathology and apoptosis induced by ischemia-reperfusion injury through adjusting the ratio of Bcl-2/Bax protein expression, thus resulting in the reduction of expression of caspase-3.

The immunohistochemical localization of pregnancy-specific beta 1-glycoprotein in postimplantation rat trophoblast.

Pregnancy-specific beta 1-glycoprotein (SP1) was identified by peroxidase-antiperoxidase (PAP) immunohistochemistry in the placenta of inbred strains of rat between 10 and 21 days of gestation. SP1 was located predominantly in basal zone trophoblast and in intravascular trophoblast of decidual vessels, but it was absent from labyrinthine trophoblast. No perivascular cells were identified by SP1 staining, but occasional clusters of interstitial trophoblast stained for SP1. The results suggest that basal and labyrinthine trophoblast are functionally different.