Human Cytomegalovirus (HCMV) infection was not correlated with overall survival in glioblastomas.

There were many arguments about the presence of HCMV (Human Cytomegalovirus) in malignant gliomas. This study was to investigate the presence and prognostic value of HCMV in glioblastomas. 68 patients including 64 primary glioblastomas and 4 secondary glioblastomas were involved in this study. Immunofluorescence was adopted for detecting glycoprotein B (gB) and glycoprotein H (gH) of HCMV's in glioblastoma tissues. Kaplan-Meier Analysis and Chi Square were used to evaluate patients' survival and the association between HCMV infection and patients' characteristics respectively. We found that the presence rate of gB and gH were 48.5% (33/68) and 42.6% (29/68) in glioblastomas respectively. The co-occurrence of gB and gH was 30.8%, and the presence rates of either gB or gH in glioblastomas was 60.3%. While IDH R132H mutations were significantly correlated with a better clinical outcome (p=0.006), the presence of neither gB (p=0.551) nor gH (p=0.871) had prognostic values. Furthermore, there was no significant association between the presence of HCMV and gliomas' characteristics, neither with patients' age, gender, KPS, IDH mutations nor PTEN loss. In conclusion, our results supported the fact that HCMV was detected in glioblastomas. However, no predictive value of HCMV was observed, the treatment of glioblastomas targeting HCMV was needed to be revalued by studied again.

Gene cloning of porcine adiponectin gene from adipose tissue and construction of its eukaryotic expression vector.

To clone adiponectin (ADPN) gene from Shaziling porcine adipocyte and construct its eukaryotic expression vector, total RNA was extracted from subcutaneous fatty tissue. One pair of specific primers was designed by Primer 5.0 software according to the sequence of ADPN gene of porcine available in GenBank. The ADPN gene was amplified by PCR from cDNA and cloned into pMD18-T vector to construct recombinant clonal vector pMD-ADPN, sequenced and analysed. A recombinant expression plasmid pPICZaA-ADPN was constructed by subcloning the cloned ADPN gene into the linearized pPICZaA vector. Then, the plasmid pPICZaA-ADPN was expressed in Pichia pastoris (GS115) by electrotransformation. Western blot and Bradford analysis were used to determine the target protein induced by methanol. Results showed that the genome size of ADPN was 732 bp and encoded 244 amino acid, the nucleotide sequence of ADPN shared 100% identity with that of porcine available in GenBank. Western blot and Bradford analysis showed that the recombinant ADPN was expressed in GS115 correctly and has certain immune activity. The expression level of ADPN was 28.5 µg/ml. In conclusion, the recombinant ADPN could express in eukaryotic expression vector pPICZaA-ADPN constructed in this study effectively.

The RHOX homeobox gene cluster is selectively expressed in human oocytes and male germ cells.

STUDY QUESTION: What human tissues and cell types express the X-linked reproductive homeobox (RHOX) gene cluster? SUMMARY ANSWER: The RHOX homeobox genes and proteins are selectively expressed in germ cells in both the ovary and testis. WHAT IS KNOWN ALREADY: The RHOX homeobox transcription factors are encoded by an X-linked gene cluster whose members are selectively expressed in the male and female reproductive tract of mice and rats. The Rhox genes have undergone strong selection pressure to rapidly evolve, making it uncertain whether they maintain their reproductive tissue-centric expression pattern in humans, an issue we address in this report. STUDY DESIGN, SIZE, DURATION: We examined the expression of all members of the human RHOX gene cluster in 11 fetal and 8 adult tissues. The focus of our analysis was on fetal testes, where we evaluated 16 different samples from 8 to 20 weeks gestation. We also analyzed fixed sections from fetal testes, adult testes and adult ovaries to determine the cell type-specific expression pattern of the proteins encoded by RHOX genes. PARTICIPANTS/MATERIALS, SETTING, METHODS: We used quantitative reverse transcription-polymerase chain reaction analysis to assay human RHOX gene expression. We generated antisera against RHOX proteins and used them for western blotting, immunohistochemical and immunofluorescence analyses of RHOXF1 and RHOXF2/2B protein expression. MAIN RESULTS AND THE ROLE OF CHANCE: We found that the RHOXF1 and RHOXF2/2B genes are highly expressed in the testis and exhibit low or undetectable expression in most other organs. Using RHOXF1- and RHOXF2/2B-specific antiserum, we found that both RHOXF1 and RHOXF2/2B are primarily expressed in germ cells in the adult testis. Early stage germ cells (spermatogonia and early spermatocytes) express RHOXF2/2B, while later stage germ cells (pachytene spermatocytes and round spermatids) express RHOXF1. Both RHOXF1 and RHOXF2/2B are expressed in prespermatogonia in human fetal testes. Consistent with this, RHOXF1 and RHOXF2/2B mRNA expression increases in the second trimester during fetal testes development when gonocytes differentiate into prespermatogonia. In the human adult ovary, we found that RHOXF1 and RHOXF2/2B are primarily expressed in oocytes. LIMITATIONS, REASONS FOR CAUTION: While the average level of expression of RHOX genes was low or undetectable in all 19 human tissues other than testes, it is still possible that RHOX genes are highly expressed in a small subset of cells in some of these non-testicular tissues. As a case in point, we found that RHOX proteins are highly expressed in oocytes within the human ovary, despite low levels of RHOX mRNA in the whole ovary. WIDER IMPLICATIONS OF THE FINDINGS: The cell type-specific and developmentally regulated expression pattern of the RHOX transcription factors suggests that they perform regulatory functions during human fetal germ cell development, spermatogenesis and oogenesis. Our results also raise the possibility that modulation of RHOX gene levels could correct some cases of human infertility and that their encoded proteins are candidate targets for contraceptive drug design.

Distal trisomy of 10q with distal monosomy of 15q due to a paternal translocation.

Distal trisomy of 10q is a rare chromosomal abnormality. Distal deletions of the terminal long arm of chromosome 15 have rarely been described. We report on a male infant with low birth weight and microcephaly, a flat face with a spacious forehead, low-set ears, blepharophimosis, microphthalmia, a small nose, and a depressed nasal bridge. Microarray comparative genomic hybridization identified that he had the karyotype 46, XY, der (15) t (10;15) (q25.2;q26.2) pat, with chromosomal breakpoints at 10q25.2 and 15q26.2. This male neonatal case had an unbalanced translocation inherited from his father who was a balanced carrier with the karyotype 46, XY, t (10;15) (q25;q26). The neonate had a partial trisomy of the long arm of chromosome 10 with a partial monosomy of distal 15q. The clinical features were in agreement with previous descriptions and allowed us to propose a growth retardation phenotype for this neonate case.

Effects of high pressure on the luminescent properties of nanocrystalline and bulk Y2O3:Eu3+.

High pressure-induced spectral changes in a 20-nm cubic nanocrystalline yttria doped with europium and its corresponding bulk were studied in the range of 550-750 nm, corresponding to the 5D0 --> 7Fd (J = 0-4) transitions. The results demonstrate that the bulk Y2O3 underwent phase transition from the cubic phase to the monoclinic phase as the pressure increased to 15 GPa, while the 20-nm nanocrystals did not. This can be concluded from the fact that the 5D0 --> 7F0 line and the three 5D0 --> 7F1 sublines originating from the cubic phase disappeared, while another group of 5D0 --> 7F0 and 5D0 --> 7F1 lines appeared. In addition, the relative intensity of the peak around 630 nm to that around 611 nm varied obviously as the pressure surpassed 15 GPa. The variations in the nanocrystals were more sluggish in comparison to those in the bulk, indicating that the nanocrystalline yttria had improved compressibility, which is attributed to an increased surface energy in nanocrystals. The local environment surrounding luminescent Eu3+ in the nanocrystals and the bulk both became more disordered with the increase of the pressure. The phase transition from the cubic to the monoclinic is irreversible.

Recombinant enterokinase light chain with affinity tag: expression from Saccharomyces cerevisiae and its utilities in fusion protein technology.

Enterokinase and recombinant enterokinase light chain (rEK(L)) have been used widely to cleave fusion proteins with the target sequence of (Asp)(4)-Lys. In this work, we show that their utility as a site-specific cleavage agent is compromised by sporadic cleavage at other sites, albeit at low levels. Further degradation of the fusion protein in cleavage reaction is due to an intrinsic broad specificity of the enzyme rather than to the presence of contaminating proteases. To offer facilitated purification from fermentation broth and efficient removal of rEK(L) after cleavage reaction, thus minimizing unwanted cleavage of target protein, histidine affinity tag was introduced into rEK(L). Utilizing the secretion enhancer peptide derived from the human interleukin 1 beta, the recombinant EK(L) was expressed in Saccharomyces cerevisiae and efficiently secreted into culture medium. The C-terminal His-tagged EK(L) was purified in a single-step procedure on nickel affinity chromatography. It retained full enzymatic activity similar to that of EK(L), whereas the N-terminal His-tagged EK(L) was neither efficiently purified nor had any enzymatic activity. After cleavage reaction of fusion protein, the C-terminal His-tagged EK(L) was efficiently removed from the reaction mixture by a single passage through nickel-NTA spin column. The simple affinity tag renders rEK(L) extremely useful for purification, post-cleavage removal, recovery, and recycling and will broaden the utility and the versatility of the enterokinase for the production of recombinant proteins.

A case of persistent endometriosis after total hysterectomy with both salpingo-oophorectomy managed by radiation therapy.

Persistent endometriosis after total hysterectomy and both salpingo-oophorectomy (TH with BSO) is a rare condition and the etiology is uncertain. The exact incidence of persistent endometriosis after definitive surgery is not known. In addition, the treatment of persistent endometriosis after complete surgical excision is controversial. We report a case of persistent endometriosis with vaginal and sigmoid-colonic invasion after TH with BSO. The lesions were not responsive to hormonal therapy. The patient was managed successfully by therapeutic pelvic radiation.

Age estimation using a modified HPLC determination of ratio of aspartic acid in dentin.

Diastereomeric dipeptides were derived from the amino acid enantiomers in dentin by O-phthalaldehyde-N-acetyl-L-cysteine. The products were separated using high performance liquid chromatography (HPLC) and detected by fluorescence detector. A short analysis time (total analysis time was 15 min, including retention time, sample derivatization time and column regeneration time) was used. The sensitivity of detection was 1 pmol and high resolution (Rs = 1.5) was reached. We determined the D/L ratio of aspartic acid in dentin of 28 first premolars. The correlation value between the D/L ratio of aspartic acid and actual age was 0.9887; errors of +/- 1 year accounted for 46.4%, and no error exceeded 5 years.

Studies on the crystal structure of [L-Met]B0 bovine insulin at 3.0A resolution.

Based on the crystal symmetry of [L-Met]B0 bovine insulin (LMBBI) and the fundamental theory of the molecular packing method, the scheme for the determination of the position and orientation of the molecules by only using one-dimensional rotation and one-dimensional translation was chosen to be used, and therefore the calculation of the rotation function of the molecular replacement method and the refinement of the rotational and translational parameters by using the R-factor search method were simplified greatly. After the preliminary refinement by using the macromolecular rigid body refinement technique, the molecular model was further refined and adjusted by using the energy-minimizing stereochemical-restrained least squares refinement technique assisted by the manual revision on the difference Fourier maps. The L-Met residues on the N-terminus of the B-chain appeared clearly on the final electron density map.

Sex diagnosis of Chinese skulls using multiple stepwise discriminant function analysis.

Sixty Chinese skulls (30 males and 30 females) from Liaoning Province of the People's Republic of China were used in this study. Forty-one variables on each skull were measured and one group of 14 and a second group of 5 variables were selected from all the variables by applying multiple stepwise regression on a computer. Discriminant equations for the 14 and 5 variables for sex diagnosis have been obtained and these variables are highly significant. The discriminant rate for the group of 5 variables resulted in accurate sex determination in 96.7% of cases. For the group of 14 variables there was 100% success rate.

Age determination of the molars.

It has generally been acknowledged that the estimation of age may have a particular medico-legal significance, and that dental methods can be exceptionally useful (Cameron and Sims, 1973; Song and Jia, 1986a). The authors have studied the method for age estimation by using the theory of quantification (Type 1) and multiple stepwise regression analysis (Song and Jia, 1987, 1989). It is not uncommon in the practice of forensic medicine to find instances in which some teeth are missing, thereby adding to the difficulties in age estimation. Thus alternative techniques have to be used. This paper discusses a method of age estimation using only the molars in 495 females living in the Liaoning Province of the People's Republic of China.

The estimation of tooth age from attrition of the occlusal surface.

Age estimation in unidentified bodies is inaccurate. Usually only a broad range of ages, such as 20-30 years or 30-50 years, can be given, especially when postmortem change has occurred. Thus there is a real need in routine forensic practice for greater accuracy. Takei (1970, 1981), looked at the relationship between teeth and age by using the 'Theory of Quantification'. Song and Jia (1987), extended the use of this technique, adding multiple stepwise regression analysis, to the statistical examination of data derived from the study of attrition of the occlusal surfaces of teeth. The computer model thus derived provides a basis for an accurate assessment of age from qualitative material. This paper discusses the technique and is based upon the findings made during a study of the degree of occlusal attrition in 880 people, living in rural and urban areas of the Liaoning province of the People's Republic of China.