DNA microarrays have been acknowledged to represent a promising approach for the detection of viral pathogens. However, the probes designed for current arrays could cover only part of the given viral variants, that could result in false-negative or ambiguous data. If all the variants are to be covered, the requirement for more probes would render much higher spot density and thus higher cost of the arrays. Here we have developed a new strategy for oligonucleotide probe design. Using type I human immunodeficiency virus (HIV-1) tat gene as an example, we designed the array probes and validated the optimized parameters in silico. Results show that the oligo number is significantly reduced comparing with the existing methods, while specificity and hybridization efficiency remain intact. The adoption of this method in reducing the oligo numbers could increase the detection capacity for DNA microarrays, and would significantly lower the manufacturing cost for making array chips.

OBJECTIVETo investigate the effects of the hepatitis B virus (HBV) P22e protein on the apoptosis of human hepatocellular carcinoma HepG2 cells.

METHODSHepG2 cells were transfected with recombinant plasmid pEGFP-HBVP22e and exposed to Act-D and tumor necrosis factor alpha (TNFalpha) treatment to induce cell apoptosis. Flow cytometry was performed to determine the proportion of cells containing sub-G1 DNA to represent the number of apoptotic cells. Laser scanning confocal microscopy was used to observe the nuclear alterations in the apoptotic cells.

RESULTSHepG2EGFP-C2HBVP22e cell strain showed a much delayed apoptosis as well as obviously lowered apoptotic rate in comparison with the HepG2 strain (P<0.01).

CONCLUSIONThe introduction and expression of extraneous gene HBVP22e significantly inhibits the apoptosis of HepG2 cells.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Humans ; Transfection ; Viral Core Proteins ; metabolism

OBJECTIVETo observe the clinical effects of using the Chinese Shang Ring in circumcision children with either phimosis or redundant prepuce, and to investigate its superiority over the similar devices available.

METHODSA total of 824 children with phimosis or redundant prepuce underwent circumcision with the Shang Ring. The clinical data were assessed concerning the duration of the procedure, incidence of post-operative complications, time of recovery and appearance of the penis.

RESULTSThe procedure duration was (2.6 +/-1.2) min, and the complications included infection in 4 (0.6%), edema in 21 (3.2%), delayed removal of the ring in 10 (1.5%), redundant and asymmetric mucosa attributable to performance in 6 (0.9%) of the cases. The wounds healed and the rings were removed at 13.4 +/- 5.8 days after circumcision, with well-smoothed incision and good cosmetic results.

CONCLUSIONChild circumcision with the Chinese Shang Ring is easy and simple in performance, with less operative time, fewer complications and better cosmetic results.

Adolescent ; Child ; Child, Preschool ; Circumcision, Male ; instrumentation ; methods ; Humans ; Male ; Penis ; surgery ; Phimosis ; surgery ; Treatment Outcome

To establish methods and requirements for quality control of rhLFA3-IgG1, biological potency of rhLFA3-IgG1 was determined by CD2 molecule competitive binding assay on Jurkat cell surface. Purity of rhLFA3-IgG1 was analyzed by SEC-HPLC and IEC-HPLC. Peptide mapping was preformed by tryptic digestion and RP-HPLC after sample reduced and carboxymethylation by DTT and indoacetic acid, respectively. CHO host cell protein and Protein A residual were detected by ELISA separately. The quality control methods and requirements, such as biological potency, the physical-chemical characteristic of rhLFA3-IgG1 had been established. The methods and requirements for quality control of rhLFA3-IgG1 showed advantages of assuring the products safety and efficacy, which can be used for routine quality control of rhLFA3-IgG1.
Binding, Competitive ; Biotechnology ; methods ; CD2 Antigens ; metabolism ; CD58 Antigens ; biosynthesis ; chemistry ; Chromatography, High Pressure Liquid ; Humans ; Immunoglobulin G ; biosynthesis ; chemistry ; Jurkat Cells ; Molecular Weight ; Peptide Mapping ; Quality Control ; Recombinant Fusion Proteins ; biosynthesis ; chemistry

OBJECTIVETo test whether nuclear factor kappa B plays an important role in the apoptosis-inhibitory effect of hepatitis B virus (HBV) P22(e) protein.

METHODSHepG2 cells were transfected with recombination plasmid pEGFP-HBVP22(e). The Act-D and TNF alpha were used to induce apoptosis. NF-kappa B inhibitor ALLN were used to inhibit the signaling pathway. The activation of NF-kappa B was EMSA, and the nulear translocation of NF-kappa B was determined by immuno-staining.

RESULTSLaser scanning confocal microscopy and EMSA indicated that HBV P22(e) protein enhanced the nuclear translocation of NF-kappa B after apoptosis induction. ALLN treatment inhibited the nuclear translocation of NF-kappa B, and blocked the apoptosis-inhibiting effect of HBV P22(e) protein.

CONCLUSIONThis study indicates that HBV P22(e) protein inhibits apoptosis of hepatocyte via the NF-kappa B signaling pathway.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B virus ; genetics ; Humans ; Leupeptins ; pharmacology ; Liver Neoplasms ; metabolism ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Plasmids ; Signal Transduction ; drug effects ; Transfection ; Viral Core Proteins ; metabolism

Objective To observe the effects of ultra-wideband (UWB) electromagnetic irradiation on the ultrastructure of pituitary or testis and the serum sex hormones level of in rats. Methods SD male rats were divided randomly into control group and irradiation groups exposed to 3×105 pulses UWB irradiation for 6,12, 24, 48 h. After exposure, the ultrastructure changes of pituitary and testis were observed by electron microscope. Serum testosterone (T) , luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Results The rat pituitary and testis were injured at the different time after exposure to UWB electromagnetic irradiation, but the damage was serious at the 24 h after exposure. In the basophilic cell of the pituitary, there were the vacuoles and lipid droplets, dilated endoplasmic reticulum (ER),exuded lymphocytes and gathered chromatin in the cell border. In the testis tissue, there were the dilated ER and gathered chromatin in the cell border of the spermatogonium, interstitial cell and sustentacular cell, some mitochondria became vacuolar and swollen in the capillary endotheliocytes and exuded lymphocytes. Serum Tlevels of the irradiation groups at 24 and 48 h were ( 1209.7±115.7 ), ( 1340.5±331.1 ) μg/L, which were significantly lower than those[(2721.8± 178.9) and ( 2820.9±321.4 ) μg/L]of control groups at 24 and 48 h (P＜0.05). There was no significant difference for LH and FSH between exposure groups and control groups. Conclusions UWB electromagnetic radiation could induce some changes in the ultrastructure of rat pituitary and testis and serum T level, which may induce the change of reproductive system.

Objective To observe the effects of ultra-wideband (UWB) electromagnetic irradiation on the ultrastructure of pituitary or testis and the serum sex hormones level of in rats. Methods SD male rats were divided randomly into control group and irradiation groups exposed to 3×105 pulses UWB irradiation for 6,12, 24, 48 h. After exposure, the ultrastructure changes of pituitary and testis were observed by electron microscope. Serum testosterone (T) , luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Results The rat pituitary and testis were injured at the different time after exposure to UWB electromagnetic irradiation, but the damage was serious at the 24 h after exposure. In the basophilic cell of the pituitary, there were the vacuoles and lipid droplets, dilated endoplasmic reticulum (ER),exuded lymphocytes and gathered chromatin in the cell border. In the testis tissue, there were the dilated ER and gathered chromatin in the cell border of the spermatogonium, interstitial cell and sustentacular cell, some mitochondria became vacuolar and swollen in the capillary endotheliocytes and exuded lymphocytes. Serum Tlevels of the irradiation groups at 24 and 48 h were ( 1209.7±115.7 ), ( 1340.5±331.1 ) μg/L, which were significantly lower than those[(2721.8± 178.9) and ( 2820.9±321.4 ) μg/L]of control groups at 24 and 48 h (P＜0.05). There was no significant difference for LH and FSH between exposure groups and control groups. Conclusions UWB electromagnetic radiation could induce some changes in the ultrastructure of rat pituitary and testis and serum T level, which may induce the change of reproductive system.

Objective To observe the outcome of percutaneous balloon mitral valvuloplasty (PBMV) in patients with rheumatic mitral valve stenosis. Methods From April 1992 to November 2008, 1768 patients underwent PBMV in our hospital. Clinical and echocardiographic follow up data were analyzed in 426 patients from April 1992 to August 1998. Left atrial pressure and the mitral valve gradient (MVG) were measured before and immediately after PBMV in all patients. Results PBMV was successful in 1748 out of 1768 patients (98.86%). Left atrial pressure decreased from (38±7) mm Hg(1 mm Hg=0.133 kPa)to (12±4) mm Hg (P<0.001), MVG decreased from (28±6) mm Hg to (8±3) mm Hg (P< 0.001) and the area of the mitral valve increased from (0.98±0.26) cm~2 to (1.97±0.39) cm~2(P< 0.001) post PBMV. The main complications included death (n=2), acute pericardial effusion (n=1), severe mitral regurgitation (n=12), cerebral embolism (n=2) and pulmonary edema (n=1). Ten years follow up was finished in 426 patients and 288 patients (67.6%) were still in NYHA class Ⅰ or Ⅱ without mitral valve replace operation or repeated PBMV, restenosis was evidenced in 140 patients (33.3%) and 31 patients dead (7.5%). Conclusion PBMV was an effective therapy option for patients with rheumatic mitral valve stenosis.

Background: and Purpose This study aimed to construct an optimal dynamic nomogram for predicting malignant brain edema (MBE) in acute ischemic stroke (AIS) patients after endovascular thrombectomy (ET). Methods: We enrolled AIS patients after ET from May 2017 to April 2021. MBE was defined as a midline shift of >5 mm at the septum pellucidum or pineal gland based on follow-up computed tomography within 5 days after ET. Multivariate logistic regression and LASSO (least absolute shrinkage and selection operator) regression were used to construct the nomogram. The area under the receiver operating characteristic curve (AUC) and decisioncurve analysis were used to compare our nomogram with two previous risk models for predicting brain edema after ET. Results: MBE developed in 72 (21.9%) of the 329 eligible patients. Our dynamic web-based nomogram (https://successful.shinyapps.io/DynNomapp/) consisted of five parameters: basal cistern effacement, postoperative National Institutes of Health Stroke Scale (NIHSS) score, brain atrophy, hypoattenuation area, and stroke etiology. The nomogram showed good discrimination ability, with a C-index (Harrell’s concordance index) of 0.925 (95% confidence interval=0.890–0.961), and good calibration (Hosmer-Lemeshow test, p=0.386). All variables had variance inflation factors of <1.5 and tolerances of >0.7, suggesting no significant collinearity among them. The AUC of our nomogram (0.925) was superior to those of Xiang-liang Chen and colleagues (0.843) and Ming-yang Du and colleagues (0.728). Conclusions Our web-based dynamic nomogram reliably predicted the risk of MBE in AIS patients after ET, and hence is worthy of further evaluation.

This study was aimed to investigate the regulative effect of ERK and p38 signal transduction pathway on cell cycle of CML. The mRNA and protein expression of ERK, p38, cyclin D(2), cyclin E and p27 (ERK and p38 were Phosph-ERK and Phosph-P38) in CML cells and K562 cell lines were detected by RT-PCR and Western blot, respectively; cell cycle was determined by FCM, and their relationship was analyzed. The results showed that the mRNA and protein expressions of ERK, p38, cyclin D(2) and cyclin E in CML cells and K562 cells increased (P<0.01) and the expression of p27 decreased (P<0.01). There was positive correlation between the protein expressions of cyclin D(2) and the protein expression of ERK, p38 and cyclin E, but there was negative correlation between the protein expressions of cyclin D(2) and the protein expression of p27. The percentage of cells in G(0)/G(1) phase was decreased and the percentage of cells in S phase was increased, there was significant difference as compared with control (P<0.05). It is concluded that increase of the mRNA expression and protein activity of ERK and p38 activate the cell cycle-regulating proteins such as cyclin D(2), cyclin E, p27 which results in shortening of G(0)/G(1) phase, switching cell to S phase through G(1)/S check point quickly and accelerating cell cycle progression and cell proliferation, and eventually leads to occurrence of CML.
Adult ; Cell Cycle ; physiology ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Male ; Middle Aged ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism