The long-term outcomes of vaginoplasty using acellular porcine small intestinal submucosa grafts in patients with Mayer-Rokitansky-Küster-Hauser syndrome: A case series.

OBJECTIVE: To investigate the long-term outcomes for Mayer-Rokitansky-Küster-Hauser syndrome (MRKH) patients undergoing vaginoplasty using acellular porcine small intestinal submucosa grafts (SIS). DESIGN: A case series. POPULATION: Seventy-eight MRKH syndrome patients and a post-SIS patient who delivered a baby following the world's first robot-assisted uterus transplantation. METHODS: Mayer-Rokitansky-Küster-Hauser syndrome patients were grouped based on the postoperative time and the diagnosis-surgery interval. Outcomes of sexual function and psychological status were assessed using the female sexual function index (FSFI), self-rating scale of body image (SSBI) and self-acceptance questionnaire (SAQ). Anatomical outcomes were measured by clinicians. MAIN OUTCOME MEASURES: The primary outcome was restoration of sexual function, defined by an FSFI score in the 'good' range. Anatomical and psychological outcomes were also analysed. RESULTS: Sexual function was restored in 42.3% (33/78) of patients and the total FSFI score was 23.44 ± 4.43. Three factors (body defect, recognition of physical appearance and willingness to change physical appearance scores) in the SSBI and two in the SAQ decreased as the postoperative time increased. Based on the interval between diagnosis and surgery, the total SSBI score was lower in the short-interval group than in the long-interval group (7.25 ± 5.55 versus 12.04 ± 10.21, p = 0.038). CONCLUSIONS: Nearly half of MRKH patients in our study had good long-term sexual function after SIS vaginoplasty. Sexual function and psychological status improved as postoperative time increased. In addition, reducing the diagnosis to surgery interval was associated with improved psychological function.

Fetal arrhythmias: prenatal evaluation and intrauterine therapeutics.

INTRODUCTION: Fetal arrhythmias are a common phenomenon with rather complicated etiologies. Debates remain regarding prenatal diagnosis and treatment of fetal arrhythmias. METHODS: The literature reporting on prenatal diagnosis and treatment of fetal arrhythmias published in the recent two decades were retrieved, collected and analyzed. RESULTS: Both fetal magnetocardiogram and electrocardiogram provide information of cardiac time intervals, including the QRS and QT durations. M-mode ultrasound detects the AV and VA intervals, fetal heart rate, and AV conduction. By using Doppler ultrasound, simultaneous recording of the atrial and ventricular waves can be obtained. Benign fetal arrhythmias, including premature contractions and sinus tachycardia, do not need any treatment before and after birth. Sustained fetal arrhythmias that predispose to the occurrence of hydrops fetalis, cardiac dysfunction or eventual fetal demise require active treatments. Intrauterine therapy of fetal tachyarrhythmias has been carried out by the transplacental route. If maternal transplacental treatment fails, intraumbilical, intraperitoneal, or direct fetal intramuscular injection of antiarrhythmic agents can be attempted. CONCLUSIONS: The outcomes of intrauterine therapy of fetal tachyarrhythmias depend on the types or etiology of fetal arrhythmias and fetal conditions. Most are curable to a transplacental treatment by the first-line antiarrhythmic agents. Fetal cardiac pacings are effective methods to restore sinus rhythm in drug-resistant or hemodynamically compromised cases. Immediate postnatal pacemaker implantation is warranted in refractory cases.

[Transformation of Protein in Sludge During High Solids Anaerobic Digestion].

Dewatered waste sludge with a total solid (TS) concentration of 12% was used for mesophilic (37℃) anaerobic digestion (AD). The biotransformation mechanism of protein and the reason for the low conversion efficiency of protein under high solids AD was investigated by analyzing the variation of protein composition in the sludge before and after AD. The results showed that the conversion rate of protein in the sludge was 34.26% after 45 days of AD. The reason for the low efficiency of protein conversion was the poor mass transfer efficiency under the condition of high solids content and the large amount of ammonia nitrogen produced with the hydrolysis. After 45 days of AD, the total ammonia nitrogen (TAN) concentration reached 1201 mg·L-1, which resulted in the inhibition of the AD process, especially the decomposition of protein. Some of the protein converted to humic acid-like and fulvic acid-like substances, which are more difficult to decompose based on the three-dimensional fluorescence spectroscopy (3D-EEM) analysis. Two-dimensional electrophoresis (2-DE)- mass spectrometry (MS) was adopted for identifying the composition of protein in sludge before and after AD. It showed that the relative molecular weight and the isoelectric point (pI) of the protein in the sludge decreased after AD and most of the proteins left in the digested sludge came from the micro-organisms. These proteins cannot be further decomposed by the microbes because of the decreased microbial metabolic capacity at the end of the AD process or lack of specific enzymes for the hydrolysis of these proteins. This ultimately resulted in the low decomposition efficiency of the total protein in the sludge.

[Construction of a High Efficiency Anaerobic Digestion System for Vinegar Residue].

The model of high solid anaerobic digestion was used by improving the degree of homogeneity of the reaction system and biogas slurry reflux to gradually increase the material load. The vinegar residue-efficient anaerobic digestion system was successfully constructed without pretreatment.The optimum anaerobic digestibility was observed when the material loading of the reaction system reached 6.15 g·(L·d)-1, when the amount of biogas produced per unit of dry material was 396 mL·g-1, and the amount of methane produced per unit of dry material was 211 mL·g-1. The degradation rate of hemicellulose reached 63.66%, which was the main reason for the improvement of anaerobic digestion performance. The degradation rates of cellulose and lignin were 21.46% and 24.43%, respectively. The lower degradation efficiency was mainly due to the complicated degradation of the benzene ring structure in lignin and hindered hydrolysis of cellulose, which had a shielding effect on cellulose degradation.

Discovery of serum proteomic biomarkers for prediction of response to moxibustion treatment in rats with collagen-induced arthritis: an exploratory analysis.

OBJECTIVE: To examine the possible impact of moxibustion on the serum proteome of the collagen-induced arthritis (CIA) rat model. MATERIALS AND METHODS: Thirty-six male Sprague-Dawley rats were included in this experiment. The CIA animal model was prepared by injection of type II bovine collagen in Freund's adjuvant on the first and seventh day. The 36 rats were randomly divided into two groups: the untreated CIA group (control), and the CIA plus treatment with moxibustion (CIA+moxi) group. Moxibustion was administered daily at ST36 and BL23 for 7, 14 or 21 days (n=12 rats each). Arthritis score was used to assess the severity of arthritis. At the end of each 7 day treatment, blood samples from the control group and the CIA+moxi group were collected. After removal of high abundance proteins from serum samples, two-dimensional gel combined with matrix-assisted laser desorption ionisation time-of-flight MS/MS (MALDI-TOF-MS/MS) techniques were performed to examine serum protein expression patterns of the CIA rat model with and without moxibustion treatment. In addition, the relevant proteins were further analysed with the use of bioinformatics analysis. RESULTS: Moxibustion significantly decreased arthritis severity in the rats in the CIA+moxi group, when compared with the rats in the CIA group 35 days after the first immunisation (p=0.001). Seventeen protein spots which changed >1.33 or <0.77 at p<0.05 using Bonferonni correction for multiple testing were found to be common to all three comparisons, and these proteins were used for classification of functions using the Gene Ontology method. Consequently, with the use of the Ingenuity Pathway Analysis, the top canonical pathways and a predicted proteomic network related to the moxibustion effect of CIA were established. CONCLUSIONS: Using the proteomics technique, we have identified novel candidate proteins that may be involved in the mechanisms of action underlying the beneficial effects of moxibustion in rats with CIA. Our findings suggest that immune responses and metabolic processes may be involved in mediating the effects of moxibustion. Moreover, periodxiredoxin I (PRDX1) and inositol 1,4,5-triphosphate receptor (IP3R) may be potential targets.

[Differential serum proteomic analysis of rheumatoid arthritis patients of cold-dampness arthralgia spasm syndrome].

OBJECTIVE: To preliminarily study the essence of rheumatoid arthritis (RA) patients of cold-dampness arthralgia spasm syndrome (CDASS) at the protein expression level. METHODS: Totally 24 RA patients were recruited from Department of Rheumatology, Affiliated Hospital of Nanjing University of Chinese Medicine from July 2009 to September 2010. They were assigned to the CDASS group and the dampness-heat arthralgia spasm syndrome (DHASS) group according to Chinese medicine syndrome typing, 12 in each group. The normal control group consisted of 12 healthy volunteers from the Health Examination Center, Affiliated Hospital of Nanjing University of Chinese Medicine. The serum proteins were compared between the CDASS group and the normal control group/the DHASS group respectively using two-dimensional gel electrophoresis. The common differential protein spots of CDASS were analyzed by mass spectrometry. The SwissProt database was inquired using Mascot Software to identify differential proteins. RESULTS: There were 81 differential protein spots between the CDASS group and the normal control group. There were 45 differential protein spots between the CDASS group and the DHASS group. Thirteen protein spots were found to be higher or lower in protein expression quantity of the CDASS group when compared with those of the other two groups. Nine differential protein spots were identified by mass spectrometry and database retrieval. It's suggested that these proteins were most likely to be related with inhibition of cellular events, such as cell proliferation, cell differentiation, and so on. CONCLUSION: 4.1 protein and DLC-1 protein were of potential significance in the diagnosis, prognostic markers, or treatment targets of RA patients of CDASS, which also provided evidence for further studies on the essence of CDASS.

Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin inhibits the proliferation of ARPE-19 cells.

BACKGROUND: The antiproliferative effect of the Hsp90 inhibitor 17-AAG (17-allylamino-17-demethoxygeldanamycin) on human retinal pigment epithelial cells is investigated. METHODS: MTT and flow cytometry were used to study the antiproliferative effects of the 17-AAG treatment of ARPE-19 cells. 2D gel electrophoresis (2-DE) and mass spectrometry were applied to detect the altered expression of proteins, which was verified by real-time PCR. Gene Ontology analysis and Ingenuity Pathway Analysis (IPA) were utilized to analyze the signaling pathways, cellular location, function, and network connections of the identified proteins. And SOD assay was employed to confirm the analysis. RESULTS: 17-AAG suppressed the proliferation of ARPE-19 cells by inducing cell cycle arrest and apoptosis. Proteomic analysis revealed that the expression of 94 proteins was altered by a factor of more than 1.5 following exposure to 17-AAG. Of these 94, 87 proteins were identified. Real-time PCR results indicated that Hsp90 and Hsp70, which were not identified by proteomic analysis, were both upregulated upon 17-AAG treatment. IPA revealed that most of the proteins have functions that are related to oxidative stress, as verified by SOD assay, while canonical pathway analysis revealed glycolysis/gluconeogenesis. CONCLUSIONS: 17-AAG suppressed the proliferation of ARPE-19 cells by inducing cell cycle arrest and apoptosis, and possibly by oxidative stress.

Identification and characterization of cul-3b, a novel hominine CUL-3 transcript variant.

AIM: To identify genes related to the human testis development by substrate hybridization technique. METHODS: A human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription reactions. The differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank database. Multi-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to determine the tissue expression profile of cul-3b. RESULTS: Cul-3b, a novel CUL-3 transcript variant, was identified. The expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ones. Cul-3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites IRESes in the 5'-UTR. These led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict circumstances. Additionally, cul-3b expressed ubiquitously in human tissues according to multi-tissue RT-PCR. CONCLUSION: Cul-3b is a novel transcript variant of CUL-3, which may be important not only for the development of human testis but also for that of other organs.

A spermatogenesis-related gene expression profile in human spermatozoa and its potential clinical applications.

Spermatogenesis is an essential stage in the human reproductive process. In a previously study aiming to determine which genes might be involved in spermatogenesis, we compared the gene expression profiles of adult and fetal testes by hybridizing cDNA probes prepared from adult and fetal testes to membranes dotted with gene clones derived from a commercial human testis library. We identified 266 differentially expressed genes that showed higher expression levels in adult testes, indicating their potential roles in spermatogenesis. In the present study, we applied the same cDNA microarray technique to the analysis of gene expression in the spermatozoa of normal fertile men and found 149 genes that were expressed at higher levels in adult testis. A further study of five sperm motility-related genes selected from this profile by real-time PCR revealed that there was significant difference in the expression levels of two genes ( TPX-1, testis-specific protein 1 and LDHC, lactate dehydrogenase C, transcript variant 1) between normal ( n=29) and motility impaired ( n=24) semen samples, indicating that these genes are involved in sperm function. Our results demonstrated that spermatogenesis-related gene profiling could help to assess sperm quality in humans, and further study of these genes will help us to elucidate the mechanisms involved in spermatogenesis and diseases relating to human infertility.