OBJECTIVETo prepare monoclonal antibodies (McAbs) against human mesenchymal stem cells (hMSCs) and to study their biological characteristics.

METHODSBALB/C mice were immunized with pooled hMSCs. McAbs were prepared by hybridoma technique and their biological characteristics were analyzed by indirect immunofluorescence, immunohistochemistry and flow cytometry.

RESULTFive hybridoma cell lines were successfully established, which secret McAbs specifically against hMSCs. Investigations showed that all these McAb reacted only to hMSCs and had no cross-reaction to other human cells, the relative affinities of 5 McAbs were 1x10(6) (ZUB1), 1x10(5) (ZUB4), 1x10(6) (ZUC3), ZUE12 (1x10(5)) and 1x10(5) (ZUF10), respectively. Isotype analysis showed that ZUB1, ZUE12, ZUF10 against the same isotype, while ZUC3, ZUB4 against other two different isotypes alone. Flow cytometric analysis showed that the positive expression rate of cultured hMSCs was 87.39% (ZUB1), 88.07% (ZUB4), 88.12% (ZUC3), 69.89% (ZUE12) and 83.67% (ZUF10).

CONCLUSIONThe prepared five McAbs can specifically react against hMSCs, which can be used for selection and study of hMCSs.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibody Specificity ; Bone Marrow Cells ; cytology ; immunology ; Fluorescent Antibody Technique ; methods ; HL-60 Cells ; Humans ; Hybridomas ; secretion ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Mesenchymal Stromal Cells ; cytology ; immunology ; Mice ; Mice, Inbred BALB C ; Rats

OBJECTIVETo construct the eukaryotic expression plasmid containing mouse vasoactive intestinal peptide(VIP) gene with biological activities.

METHODSVIP cDNA including the sequences of signal peptide was cloned from mouse thymus by RT-PCR, and then inserted into the mammalian expression vector pcDNA3.1 between Hind III and EcoR I restriction sites. COS-7 cells were transfected with pcDNA3. 1-VIP using liposome, the expression of VIP was identified by Western blot and ELISA. Supernatant of transfected cell culture was added to LPS-stimulated macrophages and the TNF-alpha production in cell medium was observed by ELISA.

RESULTSThe cloned VIP cDNA was confirmed by enzyme digestion and DNA sequencing. The expression of VIP was detected in the pcDNA3. 1-VIP transfected COS-7 cells by Western blot and ELISA. The VIP in culture supernatant potently inhibited TNF-alpha production by LPS-induced Macrophages in vitro.

CONCLUSIONThe eukaryotic expression plasmid that expresses biological active murine VIP has been constructed successfully.

Animals ; Base Sequence ; COS Cells ; Cercopithecus aethiops ; Cloning, Molecular ; DNA, Complementary ; genetics ; Eukaryotic Cells ; metabolism ; Mice ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Vasoactive Intestinal Peptide ; biosynthesis ; genetics

OBJECTIVETo investigate the efficacy of the procedure for prolapse and hemorrhoids (PPH) combined with external hemorrhoids excision in the treatment of III or IV mixed hemorrhoids.

METHODSOne hundred and twelve patients with III or IV mixed hemorrhoids admitted for surgical treatment were randomly divided into three groups: PPH 1 group (34 cases), PPH2 group (36 cases), and Milligan-Morgan group (42 cases). PPH1 group received the standard PPH operation, PPH2 received PPH and external hemorrhoids excision, and Milligan-Morgan group received Milligan-Morgan hemorrhoidectomy. Postoperative 24 h-pain index, pain index when defecating, bleeding, anal discomfort feeling , wound edema, the ability of controlling feces, operating time, hospitalization time and charges were recorded. The change of anal dynamics was detected by anorectal manometry. All the patients were followed-up for 0.5-1 year.

RESULTSThere were no significant differences among the three groups in bleeding, anal discomfort feeling, the ability of controlling feces (P>0.05). The postoperative 24 h-pain index of PPH1 group was lower than those of the other two groups (P<0.05). PPH1 group and PPH2 group were better than Milligan-Morgan group in pain index when defecating, wound edema, operating time, and hospitalization time (P<0.05). Milligan-Morgan group was better than the other two groups in postoperative urinary retention and hospital charges (P<0.05). The change of anal duct pressure of Milligan-Morgan group was less than those of the other two groups (P<0.05). Within 0.5-1.0 year follow-up, 3 patients got thrombosed external hemorrhoid in PPH1 group, 2 patients recurred and 1 patient got thrombosed external hemorrhoid in Milligan-Morgan group, no recurred patients in PPH2 group.

CONCLUSIONPPH combined with external hemorrhoid excision is a safe and effective treatment for mixed hemorrhoids, which is suitable for mixed hemorrhoids with severe external hemorrhoids.

Adult ; Aged ; Anal Canal ; surgery ; Female ; Follow-Up Studies ; Hemorrhoids ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Prolapse ; Surgical Stapling

OBJECTIVETo evaluate the factors for the conversion of transurethral resection of the prostate (TURP) to open prostatectomy and to provide clinical evidence for surgical options.

METHODSFrom January 1997 to March 2005, we performed 1 086 TURP and made retrospective analyses of 11 risk factors concerning the demographics, clinical history, laboratory data, ultrasound results, and intraoperative complications of the patients. In addition, multivariate logistic regression was used to determine those variables predicting the conversion of TURP.

RESULTSThirty-nine (3.59%) of the TURP cases required conversion, mostly because of uncontrollable hemorrhage (71.79%). Multivariate analyses showed that a prostate volume > 85.2 ml (OR = 2.568, P < 0.01), intraoperative slit of capsula prostatic (OR = 1.916, P < 0.01) and a second midstream bladder specimen (VB2) white blood cell count of the urine > 13.5/HP (OR = 1.486, P < 0.01) predicted the conversion to open prostatectomy.

CONCLUSIONBenign prostatic hyperplasia (BPH) patients with a huge prostate and those with intraoperative slit of capsula prostatic undergoing TURP are more likely to be converted. And uncontrollable hemorrhage, huge prostate and poor endoscopic vision are the major reasons for the conversion.

Aged ; Aged, 80 and over ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Postoperative Hemorrhage ; etiology ; Prostatectomy ; methods ; Prostatic Hyperplasia ; surgery ; Retrospective Studies ; Risk Factors ; Transurethral Resection of Prostate ; adverse effects ; methods ; Treatment Outcome

OBJECTIVETo compare the efficacy, tolerance and safety between oral sodium phosphate(NaP) and polyethylene glycol(PEG) on bowel preparation.

METHODSOne hundred and fifteen inpatients were randomly divided into NaP group and PEG group. The questionnaire was designed for scoring by patients and doctors regarding to tolerance, taste, side effects and cleaning degree etc.

RESULTSCompared with PEG group, NaP presented better tolerance, lower side effects and higher rate of adequate cleaning quality(P<0.05). NaP could cause electrolytic alterations, such as hyperphosphatemia, hypernatremia, hypocalcemia and hypopotassemia, but these changes were transient and without clinical significance.

CONCLUSIONSodium phosphate is safe and effective for bowel preparation, and is better than polyethylene glycol in tolerance.

Administration, Oral ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Phosphates ; therapeutic use ; Polyethylene Glycols ; therapeutic use ; Preoperative Care ; methods ; Prospective Studies ; Young Adult

Lysinuric protein intolerance (LPI) is an autosomal recessive disorder caused by SLC7A7 gene mutation and often involves severe lesions in multiple systems. Lung involvement is frequently seen in children with LPI and such children tend to have a poor prognosis. This article summarizes the clinical manifestations and gene mutation characteristics of three children diagnosed with LPI by SLC7A7 gene analysis. All three children had the manifestations of aversion to protein-rich food after weaning, delayed development, anemia, hepatosplenomegaly, and osteoporosis, as well as an increase in orotic acid in urine. In addition, interstitial pneumonia and diffuse pulmonary interstitial lesions were observed in two children. SLC7A7 gene detection showed three pathogenic mutations in these children, namely c.1387delG(p.V463CfsX56), c.1215G>A(p.W405X) and homozygous c.625+1G>A. After a definite diagnosis was made, all three children were given a low-protein diet and oral administration of citrulline [100 mg/(kg.d)], iron protein succinylate [4 mg/(kg.d)], calcium and zinc gluconates oral solution (10 mL/day) and vitamin D (400 IU/day). In addition, patient 3 was given prednisone acetate (5 mg/day). The children had varying degrees of improvement in symptoms and signs. It is hard to distinguish LPI from urea cycle disorder due to the features of amino acid and organic acid metabolism in LPI, and SLC7A7 gene analysis is the basis for a definite diagnosis of LPI.
Amino Acid Metabolism, Inborn Errors ; genetics ; Child ; Citrulline ; Fusion Regulatory Protein 1, Light Chains ; genetics ; Humans ; Lysine ; Mutation

BACKGROUNDRandomized, controlled trials have demonstrated the superiority of sirolimus-eluting stent (SES) implantation during primary percutaneous coronary intervention (PCI), as opposed to bare-metal stents, in patients with ST-elevation myocardial infarction (STEMI). This study aimed to test the hypothesis that clinical benefits of SES treatment were independent of gender in this setting.

METHODSA total of 2042 patients with STEMI undergoing SES-based primary PCI were prospectively enrolled into Shanghai Acute Coronary Event (SACE) registry (1574 men and 468 women). Baseline demographics, angiographic and PCI features, and in-hospital and 30-day major adverse cardiac events (MACE) were analyzed as a function of gender.

RESULTSCompared with men, women were older and more frequently had hypertension, diabetes, and hypercholesterolemia. Use of platelet glycoprotein IIb/IIIa receptor inhibitor (GPI, 65.5% vs. 62.2%, P = 0.10) and procedural success rate (95.0% vs. 94.2%, P = 0.52) were similar in both genders. In-hospital death and MACE occurred in 3.8% and 7.6%, and 4.5% and 8.1% in the male and female patients, respectively (all P > 0.05). At 30-day follow-up, survival (94.3% vs. 93.8%, P = 0.66) and MACE-free survival (90.2% vs. 89.3%, P = 0.52) did not significantly differ between men and women. After adjustment for differences in patient demographics, angiographic and procedural features, there were no significant difference in either in-hospital (OR = 0.77, 95%CI of 0.48 to 1.22, P = 0.30) or 30-day mortality (OR = 1.28, 95%CI of 0.73 to 2.23, P = 0.38) between women and men.

CONCLUSIONDespite more advanced age and clustering of risk factors in women, female patients with STEMI treated by SES-based primary PCI had similar in-hospital and short-term clinical outcomes as their male counterparts.

Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Anti-Bacterial Agents ; therapeutic use ; China ; Drug-Eluting Stents ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; mortality ; therapy ; Prospective Studies ; Registries ; Sex Factors ; Sirolimus ; therapeutic use

OBJECTIVETo analyze the clinical and molecular features of a child with carnitine palmitoyltransferase 1A (CPT1A) deficiency.

METHODSClinical data of the child was collected. Blood acylcarnitine was determined with tandem mass spectrometry. DNA was extracted from the child and his parents. All exons and flanking regions of the CPT1A gene were analyzed by PCR and Sanger sequencing.

RESULTSAnalysis showed that the patient carried compound heterozygous mutations c.1787T>C and c.2201T>C of the CPT1A gene, which derived his father and mother, respectively. Both mutations were verified as novel through the retrieval of dbSNP, HGMD and 1000 genome databases. Bioinformatic analysis suggested that the mutations can affect protein function.

CONCLUSIONAcyl carnitine analysis has been the main method for the diagnosis of CPT1A deficiency. The c.1787T>C and c.2201T>C mutations of the CPT1A gene probably underlie the disease in this patient. Gene testing can provide important clues for genetic counseling and prenatal diagnosis.

Base Sequence ; Carnitine O-Palmitoyltransferase ; deficiency ; genetics ; Exons ; Female ; Humans ; Hypoglycemia ; enzymology ; genetics ; Infant ; Lipid Metabolism, Inborn Errors ; enzymology ; genetics ; Male ; Molecular Sequence Data ; Point Mutation ; Pregnancy

BACKGROUNDCurrent guidelines support primary percutaneous coronary intervention (primary PCI) as the first treatment of choice (as opposed to thrombolytic therapy) for patients with acute ST-segment elevation myocardial infarction (STEMI) especially when delivered within 12 hours of symptom onset. We aimed to evaluate the impact of different clinical pathways on reduction of reperfusion delay and subsequent improvement in outcomes in patients with STEMI.

METHODSFrom November 2005 to November 2007, 546 consecutive patients with definite STEMI, who upon arrival at the emergency room were triaged to undergo primary PCI, were included. Of them, 271 patients were brought directly to catheterization laboratory (rapid group), and 275 patients were admitted to the coronary care unit (CCU) or cardiac ward first, and then transferred to the catheterization laboratory (non-rapid group). Primary endpoint was door-to-balloon (D2B) time, and secondary endpoints included infarct size assessed by peak CK-MB level and rates of major cardiac adverse events (MACE) including death, reinfarction, or target-vessel revascularization during hospitalization and at 30-day clinical follow-up.

RESULTSBaseline clinical characteristics, angiographic features and procedural success rates were comparable between the two groups, except that more patients received glycoprotein IIb/IIIa receptor inhibitors before angiography (84.0% and 77.1, P = 0.042) and had TIMI 3 flow in the culprit vessel at initial angiogram (17.1% and 9.2%, P = 0.007) in the non-rapid group. The D2B time was shortened ((108 +/- 44) minutes and (138 +/- 31) minutes, P < 0.0001), and number of patients with D2B time < 90 minutes was greater (22.6% and 10.9%, P < 0.0001) in the rapid group. The advantages associated with rapid intra-hospital transfer were enhanced if the patients presented to the hospital at regular hours. Peak CK-MB level was significantly reduced in the rapid group. In-hospital mortality (4.1% and 5.8%) and cumulative MACE rate (7.0% and 9.8%) did not significantly differ between rapid and non-rapid groups. At 30 days, cumulative death- and MACE-free survival rates were improved in the rapid group (94.5% and 89.5%, P = 0.035; 90.1% and 84.0%, P = 0.034, respectively).

CONCLUSIONSClinical pathway with bypass of CCU/cardiac ward admission was associated with rapid reperfusion, smaller infarct size, and improved short-term survival for patients with STEMI undergoing primary PCI. In the future, it is essential to reduce the time delay for patients presenting at off-hours.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Critical Pathways ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; drug therapy ; mortality ; pathology ; therapy ; Prognosis ; Survival Analysis ; Time Factors ; Treatment Outcome

Objective: To evaluate the nutritional status by the Controlling Nutritional Status (CONUT) score and its association with the long-term prognosis in patients with acute heart failure (AHF). Methods: This prospective monocentric study consecutively enrolled patients admitted to our hospital for AHF from April 2012 to May 2016. Patients were divided into 3 groups based on the CONUT score at admission: normal (0-1), mild malnutrition (2-4) and moderate-severe malnutrition (5-12) groups. Baseline information was obtained and recorded within 24 hours after admission. All patients were followed up every 3 months by outpatient visit or telephone call until March 2019. The primary endpoint was all-cause mortality. The Kaplan-Meier survival curves and log-rank test were used to compare all-cause mortality between groups. Variables showing statistical significance in the univariate analysis were incorporated into multivariate Cox regression model to analyze the independent risk factors for all-cause mortality after discharge. Results: A total of 396 patients were enrolled in this study, including 114 patients with normal nutritional status, 200 patients with mild malnutrition and 82 patients with moderate-severe malnutrition. One hundred and fifty-eight patients died during a median follow-up of 34 (18, 46) months. The mortality was 32.4% (37/114), 39% (78/200) and 52.4% (43/82) in normal, mild malnutrition and moderate-severe malnutrition groups, respectively. The mortality was significantly higher in the moderate-severe malnutrition group than in normal nutrition group (P<0.05). However, there was no significant difference in mortality between normal and mild malnutrition group as well as between mild and moderate-severe malnutrition group (both P>0.05). Kaplan-Meier curves indicated that patients with high CONUT score group was at higher risk of all-cause mortality compared with those with low CONUT score (P=0.002). Cox proportional hazard analyses showed that the risk of all-cause mortality of moderate-severe malnutrition group was significantly higher than that of normal nutrition group (HR =1.648, 95%CI 1.021-2.660, P=0.041). Conclusions: The CONUT score of patients with AHF at admission is associated with the long-term prognosis. High CONUT score is an independent risk factor for all-cause mortality in AHF patients after discharge.
Heart Failure ; Humans ; Nutrition Assessment ; Nutritional Status ; Prognosis ; Prospective Studies ; Retrospective Studies