Objective: To investigate the expression of phosphoglycerate mutase 1 (PGAM1) in the mouse testis after exposure to single heat stress (SHS). METHODS: We randomly assigned 32 C57 male mice to an SHS (n = 16) and a control group (n = 16), the former bathed in water at 43 ℃ and the latter at 25 ℃ for 15 minutes. At 1 and 7 days after exposure, we harvested the testicular tissue for observation of the morphological changes of testicular cells by HE staining and determination of the location and expression of the PGAM1 protein by immunohistochemistry and Western blot. RESULTS: The testis volume of the mice were reduced significantly, the spermatogenic tubules were disorganized, and the cells were reduced in number after heat stress and basically disappeared after 7 days. Immunohistochemistry showed extensive expression of the PGAM1 protein in the testicular spermatogenic tubules of the SHS-exposed mice, significantly higher than in the control group at 1 day after exposure, which was down-regulated in the testis tissue at 7 days, but still markedly higher than that in the control. Western blot exhibited significantly up-regulated expression of the PGAM1 protein after heat stress compared with that in the control group. CONCLUSIONS The expression of the PGAM1 protein undergoes dynamic changes in the mouse testis after exposed to single heat stress, which is related to heat stress-induced proliferation and division of testicular spermatogenic cells.
Animals ; Heat-Shock Response ; Male ; Mice ; Phosphoglycerate Mutase ; Testis

OBJECTIVE: To identify potential mutation of PMM2 gene in an infant with congenital disorders of glycosylation type 1a (CDG-1a). METHODS: Genomic DNA was extracted from peripheral blood sample of the patient. All coding exons (exons 1-8) and splicing sites of the PMM2 gene were amplified with PCR. Potential variants were detected by direct sequencing of the PCR products and comparing the results against the ESP and SNP human gene databases. A protein BLAST system was employed to analyze cross-species conservation of the variants amino acid. A PubMed BLAST CD-search system was employed to identify functional domains damaged by variants of the PMM2 gene. Impact of potential variants was analyzed using software including PolyPhen-2 SIFT and Mutation Taster. Whole exome sequencing was used to identify additional variants of the PMM2 gene which may explain the condition of the patient. RESULTS: The child was found to carry compound heterozygous variants (c.458_462delTAAGA and c.395T>C) of the PMM2 gene, which were inherited respectively from his father and mother. The c.458_462delTAAGA has not been reported previously and may result in disruption of 10 functional domains within the PMM2 protein. The c.395T>C mutation has been recorded by a SNP database with frequency unknown. Both mutations were predicted as "probably damaging". Whole exome sequencing has identified no additional disease-causing variant which can explain the patient's condition. CONCLUSION The patient's condition may be attributed to the compound heterozygous variants c.458_462delTAAGA and c.395T>C of the PMM2 gene. Above results has facilitated molecular diagnosis for the patient.
Congenital Disorders of Glycosylation ; genetics ; Exons ; Humans ; Infant ; Mutation ; Phosphotransferases (Phosphomutases) ; genetics

Phosphoglycerate mutase 1 (PGAM1) is upregulated in many cancer types and involved in cell proliferation, migration, invasion, and apoptosis. However, the relationship between PGAM1 and prostate cancer is poorly understood. The present study investigated the changes in PGAM1 expression in prostate cancer tissues compared with normal prostate tissues and examined the cellular function of PGAM1 and its relationship with clinicopathological variables. Immunohistochemistry and Western blotting revealed that PGAM1 expression was upregulated in prostate cancer tissues and cell lines. PGAM1 expression was associated with Gleason score (P = 0.01) and T-stage (P = 0.009). Knockdown of PGAM1 by siRNA in PC-3 and 22Rv1 prostate cancer cell lines inhibited cell proliferation, migration, and invasion and enhanced cancer cell apoptosis. In a nude mouse xenograft model, PGAM1 knockdown markedly suppressed tumor growth. Deletion of PGAM1 resulted in decreased expression of Bcl-2, enhanced expression of Bax, caspases-3 and inhibition of MMP-2 and MMP-9 expression. Our results indicate that PGAM1 may play an important role in prostate cancer progression and aggressiveness, and that it might be a valuable marker of poor prognosis and a potential therapeutic target for prostate cancer.
Animals ; Apoptosis/genetics* ; Caspase 3/metabolism* ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Gene Deletion ; Gene Knockdown Techniques ; Humans ; Male ; Matrix Metalloproteinase 2/metabolism* ; Matrix Metalloproteinase 9/metabolism* ; Mice ; Mice, Nude ; Neoplasm Invasiveness/genetics* ; Neoplasm Transplantation ; PC-3 Cells ; Phosphoglycerate Mutase/genetics* ; Prostatic Neoplasms/pathology* ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; RNA, Small Interfering ; Transplantation, Heterologous ; bcl-2-Associated X Protein/metabolism*

OBJECTIVETo analyze the clinical and PMM2 gene mutation features of congenital disturbance of glycosylation caused by PMM2 gene mutation (PMM2-CDG, previously known as CDG 1a).

METHODThe clinical data of two Chinese patients who were clinically diagnosed as PMM2-CDG at neurology department of Beijing Children's Hospital in 2012 were retrospectively collected. The gene mutations were identified by Sanger sequencing.

RESULTBoth patients were female, aged 1 year and 1 month and 8 months respectively. The main clinical features of the two cases were developmental delay after birth, chronic diarrhea and metabolic acidosis, associated with elevated serum transaminases, and decreased antithrombin III activity. Physical examination showed esotropia, inverted nipples, and abnormal subcutaneous fat pads. The cranial MRI showed cerebellar atrophy. Both cases were treated with occupational therapy, physical therapy and speech therapy. The development was gradually improved but also delayed as compared with normal peers during follow-up for more than 3 years. Genetic analysis showed that patient 1 was compound heterozygous for c. 422G>A(p.Arg141His), which was reported for known pathogenic mutation, and c. 669C>A(p.Asp223Glu), was a new mutation. The patient 2 showed compound heterozygous mutation for c. 634A>G (p.Met212Val)and c. 713G>C(p.Arg238Pro), which were both new mutations.

CONCLUSIONPMM2-CDG is a rare metabolic disease, and the diagnosis should be considered in a child with developmental delay, elevated serum transaminases, decreased antithrombin III activity, inverted nipples, abnormal subcutaneous fat pads, esotropia, and cerebellar atrophy on MRI. It can be confirmed by PMM2 gene analysis.

Asian Continental Ancestry Group ; Congenital Disorders of Glycosylation ; genetics ; DNA Mutational Analysis ; Developmental Disabilities ; Female ; Genetic Testing ; Glycosylation ; Heterozygote ; Humans ; Infant ; Magnetic Resonance Imaging ; Mutation ; Phosphotransferases (Phosphomutases) ; genetics ; Retrospective Studies

The harshness of legionellosis differs from mild Pontiac fever to potentially fatal Legionnaire's disease. The increasing development of drug resistance against legionellosis has led to explore new novel drug targets. It has been found that phosphoglucosamine mutase, phosphomannomutase, and phosphoglyceromutase enzymes can be used as the most probable therapeutic drug targets through extensive data mining. Phosphoglucosamine mutase is involved in amino sugar and nucleotide sugar metabolism. The purpose of this study was to predict the potential target of that specific drug. For this, the 3D structure of phosphoglucosamine mutase of Legionella pneumophila (strain Paris) was determined by means of homology modeling through Phyre2 and refined by ModRefiner. Then, the designed model was evaluated with a structure validation program, for instance, PROCHECK, ERRAT, Verify3D, and QMEAN, for further structural analysis. Secondary structural features were determined through self-optimized prediction method with alignment (SOPMA) and interacting networks by STRING. Consequently, we performed molecular docking studies. The analytical result of PROCHECK showed that 95.0% of the residues are in the most favored region, 4.50% are in the additional allowed region and 0.50% are in the generously allowed region of the Ramachandran plot. Verify3D graph value indicates a score of 0.71 and 89.791, 1.11 for ERRAT and QMEAN respectively. Arg419, Thr414, Ser412, and Thr9 were found to dock the substrate for the most favorable binding of S-mercaptocysteine. However, these findings from this current study will pave the way for further extensive investigation of this enzyme in wet lab experiments and in that way assist drug design against legionellosis.
Computer Simulation* ; Data Mining ; Drug Delivery Systems ; Drug Design ; Drug Resistance ; Fever ; Legionella pneumophila* ; Legionellosis* ; Legionnaires' Disease ; Metabolism ; Phosphoglycerate Mutase

OBJECTIVETo study the histogenesis of giant cell tumor (GCT) and factors related to tumor recurrence, invasiveness and malignant transformation.

METHODSThe clinical features, radiologic classification, surgical approach, pathologic findings, immunophenotypes and follow-up data of 123 cases of GCT were analyzed.

RESULTSThere was a significant correlation between tumor recurrence and radiographic classification (P = 0.032), over-expression of CD147 (P = 0.034) and p53 (P = 0.005), and surgical approach (P = 0.0048) in GCT. The biologic behavior showed no correlation with intramedullary infiltration, cortical bone involvement, parosteal soft tissue extension, tumor thrombi, fusiform changes of mononuclear tumor cells, mitotic count, Ki-67 index, coagulative tumor necrosis, secondary aneurysmal bone cyst formation, and adjoining bony reaction. The positive rate of p63 in stromal cells of GCT (79.7%, 94/118) was significantly higher than that in chondroblastoma (44.7%, 21/47), osteosarcoma (22.2%, 10/45) and other giant cell-rich tumors.

CONCLUSIONSGCT is a bone tumor of low malignant potential. It is sometimes characterized by locally invasive growth, active proliferation, coagulative necrosis, secondary aneurysmal bone cyst and surrounding bony reaction. It is difficult to predict the biologic behavior of GCT. Over-expression of p53 in the tumor cells and CD147 in all components of GCT correlate with tumor invasiveness, recurrence and malignant transformation. Selection of suitable surgical approach with reference to radiologic classification is considered as an important factor in reducing the recurrence rate.

Adolescent ; Adult ; Aged ; Basigin ; metabolism ; Bone Neoplasms ; diagnostic imaging ; drug therapy ; metabolism ; pathology ; surgery ; Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Giant Cell Tumor of Bone ; diagnostic imaging ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Osteosarcoma ; pathology ; Phosphoglucomutase ; metabolism ; Radiography ; Tumor Suppressor Protein p53 ; metabolism ; Young Adult

OBJECTIVETo search for novel tumor associated antigens (TAA) in esophageal squamous cell carcinoma (ESCC).

METHODSThe proteins extracted from tissues of ESCC were separated by two dimensional polyacrylamide gel electrophoresis and transferred to PVDF membrane. Sera from ESCC patients and healthy individuals were used for primary antibodies for Western blot analysis. The differential spots were excised for trypsin hydrolysis and the tryptic peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The identified TAA of ESCC was validated by immunohistochemical staining (IHC).

RESULTSSera from ESCC patients yielded multiple positive spots, and one 28 800 Da protein that exhibited positive reactivity with 60% (12/20) sera of ESCC patients and only 5% (1/20) sera of healthy controls (P<0.01). The 28 800 Da protein was identified as phosphoglycerate mutase 1 (PGAM1) by MALDI-TOF-MS. Immunohistochemical analysis showed that PGAM1 was located in both cytoplasm and nucleus, and had a higher expression in cancer tissues.

CONCLUSIONPGAM1 maybe a candidate of ESCC.

Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Esophageal Neoplasms ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Phosphoglycerate Mutase ; metabolism

Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Gastrectomy ; methods ; Histiocytic Sarcoma ; metabolism ; pathology ; surgery ; Humans ; Lymphatic Metastasis ; Male ; Phosphoglucomutase ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery

Phosphoglycerate mutase (PGAM) is a key enzyme in glycolytic pathways. With PCR technique based on an EST identified in our lab, a novel gene named SjPGAM (GenBank Accession No. EU374631) was cloned. Sequence analysis revealed that the ORF of SjPGAM gene contained 753 nucleotides, encoding 250 amino acids, and the molecular weight was about 28.26 kD. Real-time PCR analysis showed that the mRNA level of SjPGAM was much higher in the 14 days and 19 days schistosomula than other stages, suggesting that the gene was a schistosomula stage differential expression gene. The SjPGAM cDNA fragment was subcloned into an expression vector pET-28a (+) and transformed into Escherichia coli BL21 cells. In the presence of IPTG, the 31 kD fusion protein was expressed in included bodies. Western blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation. The study provides important basis for investigating the mechanism of the PGAM in the glycolytic pathways of Schistosoma japonnicum.
Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Male ; Phosphoglycerate Mutase ; genetics ; immunology ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Schistosoma japonicum ; enzymology ; genetics ; Schistosomiasis japonica ; immunology ; parasitology

PURPOSE: To evaluate whether factors related to lipid and glucose metabolism have a potential role in the progression of prostate cancer, we measured the mRNA levels of the peroxisome proliferator-activated receptor (PPAR), fatty acid elongase (ELOVL), and two glycolytic enzymes in prostate cancer (CaP) tissues. MATERIALS AND METHODS: Prostate tissues, obtained from radical prostatectomy (n=10) and transurethral resection of prostate (n=18), were quickly frozen in liquid nitrogen for RNA measurements. Transcript signals of PPAR alpha, PPAR gamma, ELOVL2, ELOVL5, phosphoglycerate kinase 1 (PgK1) and phosphoglycerate mutase 2 (PgM2) were measured using a reverse-transcription polymerase chain reaction. RESULTS: The transcript signals of PPAR alpha and PPAR gamma were down-regulated in CaP tissues. In addition, the mRNA level of PgM2 in CaP tissues was lower than that in benign prostatic hyperplasia (BPH) tissues. However, the messages for ELOVL2, ELOVL5, and PgK1 were not significantly changed. CONCLUSIONS: These results suggest that lowering of the PPARalpha, PPARgamma and PgM2 messages may be involved in aberrant and uncontrolled prostate cell growth and differentiation.
Down-Regulation* ; Glucose ; Metabolism ; Nitrogen ; Peroxisome Proliferator-Activated Receptors ; Peroxisomes* ; Phosphoglycerate Kinase ; Phosphoglycerate Mutase* ; Polymerase Chain Reaction ; PPAR alpha ; PPAR gamma* ; Prostate* ; Prostatectomy ; Prostatic Hyperplasia ; Prostatic Neoplasms* ; RNA ; RNA, Messenger ; Transurethral Resection of Prostate