r study.

Animals ; Brain Abscess ; etiology ; Encephalitis ; etiology ; Humans ; Infant, Newborn ; Leukomalacia, Periventricular ; etiology ; Multiple Sclerosis ; etiology ; Nervous System Diseases ; etiology ; Signal Transduction ; Toll-Like Receptors ; chemistry ; physiology

BACKGROUNDBoth p16(INK4) and p21(Waf1) are tumor suppressors with similar biological functions in the regulation of cellular senescence. Previous reports showed that p16(INK4) could be activated by p21(Waf1) through transcriptional factor Sp1 in HeLa cells. This study was undertaken to determine the effects of p16(INK4) on the expression and functions of p21(Waf1).

METHODSHuman diploid fibroblast 2BS cells were stably transfected with sense (2BS/p16(INK4)), antisense p16(INK4) (2BS/asp16(INK4)) or empty vector (2BS/neo). Then they were assayed by reverse-transcription polymerase chain reaction (RT-PCR), fluorescence activated cell sorting (FACS) and Western blot.

RESULTS2BS/p16(INK4) cells exhibited cell cycle arrest in both G1 and G2/M phases. Endogenous p21(Waf1) protein levels increased twofold in the 2BS/p16(INK4) cells, but not decreased in the 2BS/asp16(INK4) cells. p21(Waf1) mRNA levels were not affected in neither 2BS/p16(INK4) nor 2BS/asp16(INK4) cells.

CONCLUSIONp16(INK4) may play an important role in the regulation of cellular senescence by modulating the p21(Waf1) protein level via the posttranscriptional mechanism.

Cell Cycle ; Cells, Cultured ; Cellular Senescence ; Cyclin-Dependent Kinase Inhibitor p16 ; physiology ; Cyclin-Dependent Kinase Inhibitor p21 ; physiology ; Fibroblasts ; metabolism ; Humans ; Transcription, Genetic

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

OBJECTIVETo investigate the associations between periodontal diseases, presence of Porphyromonas gingivalis (P. gingivalis), serum levels of pro-inflammatory mediators and preterm low birth weight (PLBW).

METHODS60 women (30 PLBW and 30 healthy women), were recruited after postpartum within 3 days in this case-control study. Periodontal measurements including plaque index (PI), bleeding index (BI), probing pocket depth (PD) and clinical attachment loss (CAL) were assessed. The subgingival plaque was collected before periodontal examination and analysed by polymerase chain reaction (PCR) for the presence of the 16S rRNA gene specific to P. gingivalis, while the venous and umbilical cord blood specimens collected were determined by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe PLBW cases had a poorer oral conditions and the presence of P. gingivalis was found in a higher proportion in the PLBW than the healthy pregnant (56.7% vs. 30.0%, P < 0.05). Both of the presence of periodontitis and P. gingivalis have been found to be associated weakly with a shorter gestational age and a lower birth weight (P < 0.05). The levels of IL-1beta, IL-6 and prostaglandin-E2 (PGE2) in both of the blood samples showed higer levels between the PLBW and normal groups (P < 0.01). The IL-1beta and PGE2 levels in maternal serum were higher with a severe periodontal disease in the PLBW group (P < 0.01).

CONCLUSIONThere may be a possible link between periodontal diseases and PLBW.

Adult ; Case-Control Studies ; Dental Plaque ; Dental Plaque Index ; Female ; Humans ; Infant, Low Birth Weight ; Infant, Newborn ; Interleukin-6 ; Periodontal Diseases ; Periodontitis ; Porphyromonas gingivalis ; Pregnancy ; Premature Birth ; RNA, Ribosomal, 16S

OBJECTIVETo explore the feasibility and therapeutic effect of thinned posterior tibial artery free perforator flap for the reconstruction of soft tissue defects at dorsum of hands.

METHODSSix fresh adult lower limbs specimens were injected with red latex via arterial cannula and dissected. The number, distribution, branches, and outer diameter of posterior tibia artery perforators were observed. Based on the anatomic study, the perforator flaps were designed to reconstruct soft tissue defects at dorsum of hands and wrists. The redundant fat on the flaps was removed, but preserving the nutrient vascular system. 11 flaps were used with the size ranging from 2 cm x 5 cm to 10 cm x 14 cm.

RESULTS43 skin perforators of posterior tibial artery were observed in six lower limbs, 29 perforators with the outer diameter is greater than 0.5 mm when they threading over the deep fascia plane, on average every 4.8 bundles of sides. The mean outside diameter of perforating artery is (1.8 +/- 0.5) mm, and the length is (44 +/- 15) mm. 6 perforators were founded both in the second and fifth zone which could be used for anastomosis for its better diameters. All flaps survived completely without any complication at donor sites. 7 cases were followed up for 3-12 months. Both satisfactory functional and cosmetic results were achieved with a soft and thinned appearance.

CONCLUSIONSThe thinned posterior tibial artery free perforator flap has a reliable blood supply and good appearance. It is very suitable for the reconstruction of small or medium-sized defects at the dorsum of hands and wrists.

Adolescent ; Adult ; Female ; Hand Injuries ; surgery ; Humans ; Lower Extremity ; anatomy & histology ; blood supply ; Male ; Middle Aged ; Perforator Flap ; blood supply ; Reconstructive Surgical Procedures ; methods ; Tibial Arteries ; anatomy & histology ; transplantation ; Young Adult

Objective To develop appropriate evaluation methods of local basic public health services which are suitable to county level and above.Methods Data on basic public health services of 1 1 cities in Zhejiang province in 201 2 was evaluated by different evaluation methods including weighted synthetic scored method,weighted synthetic index method, Weighted Technique for Order Preference by Similarity to Ideal Solution (Topsis ) and Weighted Rank -Sum Ratio (RSR).The consistency of evaluation results were tested by Kendall's coefficient of concordance W test.Combination evaluation was conducted to evaluate four single synthetic evaluation results through average method,weighted average combination evaluation method and hierarchical clustering analysis.Results Different synthetic evaluation methods had different evaluation results.However,in the order,the top two were all Hangzhou and Ningbo.Kendall's W test showed good consistence of four evaluation results.Rank of 1 1 cities were Hangzhou,Ningbo,Shaoxing,Jiaxing,Huzhou, Taizhou,Jinhua,Zhoushan,Lishui,Wenzhou and Quzhou based on combination evaluation value by average method, which was the same to the rank based on weighted average combination evaluation result.Eleven cities could be classified into four categories by hierarchical clustering analysis with statistical significance (P <0.01 ):Excellent (Hangzhou, Ningbo),Good (Huzhou,Jiaxing,Shaoxing),Middle (Zhoushan,Jinhua,Taizhou)and Poor (Wenzhou,Quzhou, Lishui).Conclusion These four synthetic evaluation methods used in this study are all suitable to county level and above in basic public health services evaluation.Various synthetic evaluation methods could be used in practice with combination evaluation of various evaluation results.Average method which is convenient and accurate is preferred when consistency of various synthetic evaluation results was testified.Hierarchical clustering analysis could be used for combination evaluation when no precise rank is needed.

Objective To investigate the relationship between single nucleotide polymorphisms of tumor suppressor gene PTEN and nasopharyngeal carcinoma in Jiamusi Han population. Methods The blood samples of 132 patients with naso-pharyngeal carcinoma(nasopharyngeal carcinoma group)and 73 healthy people(control group)were selected from September 2008 to January 2018 in the First Affiliated Hospital of Jiamusi University. The whole genome DNA was extracted,and the pol-ymorphisms of rs532678 and rs701848 were detected by restriction fragment length polymorphism analysis. The relationship be-tween the polymorphism of PTEN gene and nasopharyngeal carcinoma was analyzed. Results The genotype and allele frequen-cy distributions of rs532678 and rs701848 loci were in line with the Hardy-Weinberg genetic balance law in the two groups (P > 0. 05). The genotypic frequency of CC,CT and TT at the rs532678 locus of PTEN gene in the control group was 0. 630, 0. 342 and 0. 027 respectively;and the allele frequency of C and T was 0. 801 and 0. 198 respectively. The genotypic frequency of CC,CT and TT at the rs532678 locus of PTEN gene in the nasopharyngeal carcinoma group was 0. 716,0. 265 and 0. 015 re-spectively;and the allele frequency of C and T was 0. 852 and 0. 147 respectively. There was no significant difference in geno-type distribution and allele frequency distribution at the rs532678 locus of PTEN gene between the two groups(P > 0. 05). The genotypic frequency of CC,CT and TT at the rs701848 locus of the PTEN gene in the control group was 0. 657,0. 342 and 0. 000 respectively;and the allele frequency of C and T was 0. 828 and 0. 171 respectively. The genotypic frequency of CC,CT and TT at the rs701848 locus of PTEN gene in the nasopharyngeal carcinoma group was 0. 424,0. 500 and 0. 075 respectively;and the allele frequency of C and T was 0. 674 and 0. 325 respectively. The frequencies of CT,TT genotype and T allele of rs701848 locus in the nasopharyngeal carcinoma group were significantly higher than those in the control group(P < 0. 05). The frequencies of CC genotype and C allele in the nasopharyngeal carcinoma group were significantly lower than those in the control group(P < 0. 05). The individual with CT + TT genotype at the rs701848 locus of PTEN gene had higher risk for naso-pharyngeal carcinoma(P < 0. 05,OR = 2. 606,95％ confidence interval:1. 439 - 4. 720). The risk for nasopharyngeal carcino-ma in the individual with CT + TT genotype was 2. 606 times as much as the individual carrying CC genotype. Conclusion The rs532678 polymorphism of PTEN gene is not associated with the susceptibility to nasopharyngeal carcinoma. The polymor-phism of rs701848 locus is associated with the susceptibility to nasopharyngeal carcinoma. The individual carrying CT + TT genotype has higher risk for nasopharyngeal carcinoma.

OBJECTIVETo investigate the genomic variation of Epstein-Barr virus (EBV) and its significance in nasopharyngeal carcinogenesis.

METHODSForty nasopharyngeal carcinoma (NPC) biopsy tissues were used for detection of EBV BamHI f variant and LMP1 XhoI-loss by polymerase chain reaction (PCR), nested PCR, and RFLP (restriction fragment length polymorphism). Forty-eight samples of peripheral blood mononuclear cells (PBMC) taken from apparently healthy adult individuals were used for detection of LMP1 XhoI-loss. Three samples of amplified LMP1 exon 1 DNA from B95-8 cell line and 2 NPC tissues (one having XhoI-loss and the other having Wt-XhoI/XhoI-loss) were sequenced.

RESULTSThirty out of the 40 NPC cases (30/40, 75%) harbored EBV BamHI f variant and the remaining 10 (10/40, 25%) harbored BamHI F prototype. Thirty out of the 39 NPCs (30/39, 76.9%) showed single EBV LMP1 XhoI-loss, 7 (7/39, 18.0%) showed single LMP1 Wt-XhoI (presence of a XhoI site in exon 1 of LMP1 gene, as in B95-8 cell line), and 2 (2/39, 5.1%) showed both LMP1 Wt-XhoI and XhoI-loss. Thirty-eight of the 39 NPCs (97.4%) showed EBV LMP1 XhoI-loss or/and BamHI F variant. In the NPC tissue (1 case only) showing the prototype of Wt-XhoI/BamHI "f", there were several base substitutions, including 5 missense mutations and 2 silent mutations present in LMP1 exon 3, on DNA sequencing. On the other hand, 10 out of the 48 samples of PBMC taken from apparently healthy individuals could be amplified successfully by nested PCR for detection of LMP1 XhoI site. All of these 10 samples carried the prototype of EBV LMP1 Wt-XhoI.

CONCLUSIONSThe majority of EBV present in neoplastic cells of NPC is of BamHI "f" variant and/or possesses LMP1 XhoI-loss, as compared with that in healthy individuals. This genomic variation of EBV may bear some roles in the development and progression of NPC.

Adult ; Aged ; Binding Sites ; genetics ; DNA, Viral ; genetics ; metabolism ; Deoxyribonuclease BamHI ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Female ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Mutation ; Nasopharyngeal Neoplasms ; virology ; Sequence Deletion ; Viral Matrix Proteins ; genetics

The Pichia pastoris strain GS115-PreS could produce a high expression level of full-length PreS protein that secreted to the supernatant after methanol induction in the fermentation. The Western blot analysis showed a single band with expected molecular mass of 48kD and that the major component of the particles was the full-length PreS protein (PreS1 + PreS2 + S) and small envelope protein (S) of 48 and 28 kD, respectively. Electron microscopy image showed PreS particles with 30 nm in diameter. The supernatants of the fermentation were desalted and concentrated. Purified PreS protein was obtained by DEAE-SFF anion exchange column chromatography and the PreS particles were obtained by ultracentrifugation and sucrose density gradient. The ELISA assay results proved that both full-length PreS protein and particles showed high immunogenicity and specificity. P/N ratio further demonstrated that the immunogenicity of the particles is higher than the full-length PreS protein.
Hepatitis B Surface Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; immunology ; Humans ; Pichia ; genetics ; metabolism ; Protein Precursors ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification ; Viral Envelope Proteins ; biosynthesis ; genetics