Lactose was shown to no less competent than Isopropyl-?-D-thiogalactoside (IPTG) in inducing the expression of the ENHANCIN coding gene from Helicoverpa armigera granulosis virus in Eswcherichia coli BL21 (DE3) regulated by a T7 promoter, since the lactose induction could lead to an ENHANCIN band no smaller than the one in IPTG induction on the SDS-PAGE gel. This would decrease the cost of the large-scale ENHANCIN production. The lactose concentration was optimized at 2.2% - 2.5% (w/v) . Different treatments on the lactose sterilization showed that lactose steam- sterilized in 116. 5℃ for 15min could lead to the ENHANCIN production. The convenience and the relatively low cost in its" operation could further decrease the cost of the ENHANCIN production.

Objective To investigate the role of Glial cells missing 2 (Gcm2) in pathogenesis of hypoparathyroidism by knocking out Gcm2 gene in adult mice.Methods Tamoxifen was used to induce conditional knock-out of Gcm2 gene in Gcm2E2fl/flCre-ER mice.Genotypes of knock-out mice were identified by PCR.The protein expression level of Gcm2 was measured by Western blotting.The serum calcium and phosphorus were detected by the calcium and phosphorus assay kits, and the serum parathyroid hormone (PTH) level was detected by ELISA.Parathyroid cell proliferation was tested by Ki-67 immunohistochemical assay.The mRNA expression levels of PTH and calcium sensing receptor (CaSR) were detected by Real-time PCR.Bone mineral density was detected by micro CT.Results Gcm2 gene of parathyroid was confirmed to be knocked out by PCR.Compared with wild type and solvent control groups, Gcm2 knock-out group showed markedly lower protein expression of Gcm2, notably higher serum phosphorus and lower serum calcium and PTH concentrations (all P<0.01).The proliferation of parathyroid cells in Gcm2 knock-out mice were significantly higher(both P<0.01).The mRNA levels of PTH and CaSR in parathyroid gland of the knock-out group were significantly reduced (all P<0.01).Bone mineral density was significantly higher in Gcm2 knock-out group (all P<0.01).Conclusion Knockout of Gcm2 can lead to hypoparathyroidism in adult mice, indicating that Gcm2 is probably a therapeutic target for hypoparathyroidism.

Objective To explore the characteristics of ischemia-reperfusion induced infant lung damage and the potential mechanisms of the injuried.Methods Both infant (15-21 days old) and adult (5-6 months old) rabbits were subjected to either ischemia-reperfusion or sham operation.Ischemia-reperfusion was induced by clamping the right pulmonary hilum for 1 hour and then removal of the clamp for 4 hours under anesthesia.The lung tissue were sampled for histological examination by light and electron microcopies and for biological evaluation of mitochondrial alterations.Production and expression of free radical species-hydroxyl radical (ROS-HR),malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxidase (GSH-PX),myeloid differentiation factor-88 (MyD-88),and nuclear factor-κB (NF-κB) in the lung tissue were also examined.In addition,circulating levels of interleukin-β and tumor necrosis factor-α were measured during the ischemia-reperfusion process.Results In comparison to adult lungs,the infant lungs had more increased neutrophil infiltration,edema,swelled alveolar epithelial and endothelial cells,and severer mitochondrial impairment reflected by damage of the inner membrane as well as decrease in the membrane potential after ischemia-reperfusion.The lungs in infant animals subjected to sham operation displayed higher levels of ROS-HR and MDA and lower levels of SOD and GSH-PX than those in adult controls.The lungs in infants with ischemia-reperfusion were found to further produce more ROS-HR,and MDA,and less SOD and GSH-PX than the ischemia-reperfused adult lungs.Moreover,the circulating levels of interleukin-1β and tumor necrosis factor-α were elevated during the period of ischemia-reperfusion,particularly in the infant animals,which appeared to be associated with the expression of MyD-88 and NF-κB in the lungs.Conclusion Lung ischemia-reperfusion causes more severe lung damage in infants than in adults,probably due to combination of low antioxidant capacity and overproduction of ROS in infants.

At present, the objective of cutting and pruning Cistanche deserticola is to harvest in successive years and enhance the harvesting yield and quality of C. deserticola in the process of the artificial cultivating C. deserticola. An experiment was conducted focusing on cutting and pruning C. deserticola in artificial forests of Haloxylon ammodendron drip-irrigated with saline water at the hinter-land of the Taklimakan desert, according to different growth stages and lengths. The results were following: (1) The effect of cutting on C. deserticola was similar to that of pruning, which resulted in three kinds of morphological types, not related to the bloom and size of C. deserticola. (2) The growth forms were diversified after pruning. Among them, there had sprouting new body, died or maintaining life with no sprouting, mildewed on its surface layer, etc. However, some of new bodies were sprouting from the lower part of the old body. The death rate of bloomed C. deserticola was higher than that of the underground, and the death rate of the 40 cm in stubble height for C. deserticola was higher than those with the stubble height of 20 cm and 5 cm. (3) Most of the diameter of living C. deserticola after pruning was increasing, but some of them changed little. (4) The mildew and rot of C. deserticola and the broken of the roots of the H. ammodendron and the fallen of the point of the inoculated when it was dug, which would cause the death of the C. deserticola. On the other, the yield-increasing effect and the economic benefit of the techniques of the pruning of Cistanche would need further research and evaluate. Therefore, the application of this technique needs to be cautious.
Amaranthaceae ; growth & development ; Cistanche ; growth & development ; Forests ; Fruit ; growth & development ; Plant Roots ; growth & development

OBJECTIVETo investigate the effect of chitosan on the capsule inside the expanded flap.

METHODSThe expanders were implanted in animals with the treatment of chitosan(experimental group, n = 15) or without (control group, n = 15). After taking out the expanders, the flap contraction rate was calculated. The samples were observed through HE, Masson dyeing and CD34 immunohistochemical study. The thickness of capsule inside the expanded flap was measured under microscope. The samples were also studied under electron microscope.

RESULTSThe thickness of capsule was 516.000 +/- 128.491 microm in the experimental group, and 833.000 +/- 227.379 microm in the control group (P < 0.05). The number of microvessels was 8.200 +/- 2.150 per visual in experimental group, and 7.900 +/- 1.729 per visual in control group (P > 0.05). Under the electron microscope, the rough endoplasmic reticulum (RER) in the capsule in experimental group decreased and enlarged with degranulation. The mitochondria emerged or disappeared. The number of ribosome was reduced. In the control group, the RER enlarged without degranulation, the mitochondria was intact. The number of ribosome was not reduced.

CONCLUSIONSThe chitosan can effectively reduce the contraction of expanded flap through collagen secretion of fibroblast, delaying the differentiation from fibroblast to fiber cell, inhibiting thansform from fibroblast to myofibroblast. It has no effect on the microvascular generation and expansion, so the flap blood supply will not be affected with thicker capsule.

Animals ; Chitosan ; administration & dosage ; pharmacology ; Female ; Graft Survival ; Male ; Rabbits ; Skin Transplantation ; methods ; Surgical Flaps ; Tissue Expansion

OBJECTIVETo study the pathogenesis, the diagnostic criterion and the surgical methods of congenital nasal ethmoidal sinus malformation with false triple nostrils appearance.

METHODSFrom Feb 1993 to Mar 2006, a total of 13 cases of rare congenital nasal deformity had been investigated in pathogenesis, clinical manifestations, differential diagnosis and the methods of operation. The concept of congenital nasal sinus was presented. In this series, one-stage rehabilitation was achieved by using compositive operation techniques, including excision of the sinus, reconstruction of the hatch of ethmoidal sinus, transplantation of the dorsal nasal musculoaponeurotic flap as well as the nasolabial fold flap and the reconstruction of the cartilage-muscle ring in the wing of the nose.

RESULTSThe symptom disappeared in all of the 13 cases with no morbidity. The symmetrical double sides were observed and the nasal figure was satisfied.

CONCLUSIONSBy using such compositive operation techniques, including excision of the sinus, reconstruction of the hatch of ethmoidal sinus in middle nasolabial, transplantation of the dorsal nasal musculoaponeurotic flap as well as the nasolabial fold flap and the reconstruction of the cartilage-muscle ring in the wing of the nose, one-stage rehabilitation could be reached. It was an ideal, safe and reliable method to cure this kind of rare congenital nasal deformity.

Adolescent ; Child ; Congenital Abnormalities ; diagnosis ; surgery ; Female ; Humans ; Male ; Paranasal Sinuses ; abnormalities

OBJECTIVETo study the main risk factors of low back pain of workers ina foundry factory of the automobile company using cross sectional epidemiological investigation, and to provide scientific base for preventing the disorder.

METHODSThe low back pain and work loads of 1340 workers in a foundry factory of the automobile company were investigated using questionnaire, and logistic regression analysis was used to analyze the risk factors.

RESULTSThe one-year morbidity of low back pain in workers was 58.9% the morbidities of low back pain in workers engaged in foundry, transportation and modeling were 64.6%, 64.6% and 62.5%, respectively. The lifting with squat postures, bending trunk heavily, bending trunk with twisting and moving the heavy objects were found to be the most dominant risk factors for low-back pain, the OR values were 2.085, 1.961, 1.967 and 1.956, respectively. The distributions of risk factors were different among the different jobs. The logistic regression analysis showed that moving the heavy objects, lifting with squat postures, bending trunk heavily, bending trunk with twisting existed simultaneously, also the work years and gender were the risk factors.

CONCLUSIONThe manual moving heavy objects, awkward working posture or both were the most important risk factors for low-back pain. The intervene ergonomic study should be performed in future to reduce the morbidity of low-back pain.

Adult ; Automobiles ; Cross-Sectional Studies ; Female ; Humans ; Industry ; Logistic Models ; Low Back Pain ; epidemiology ; Male ; Middle Aged ; Occupational Diseases ; epidemiology ; Risk Factors ; Surveys and Questionnaires ; Workplace ; Young Adult

OBJECTIVETo explore the molecular cytogenetic characteristics in patients with chronic lymphocytic leukemia (CLL).

METHODSInterphase fluorescence in situ hybridization (FISH) was used to detect trisomy 12, deletion of 13q14 and 17p13 in 60 patients with CLL.

RESULTSOut of the 60 patients, 41 (68.3%) had at least one kind of molecular cytogenetic aberrations. Two (3.3%) had two kinds of abnormalities. Trisomy 12 was found in 12 (20.0%) cases, 13q14 deletion in 24 (40.0%) cases and 17p13 deletion in 5 (11.7%) cases. The number of trisomy 12 cells ranged from 4.0% to 34.0%, 13q14 deletion ranged from 22.0% to 93.0% and 17p13 deletion ranged from 6.0% to 68.0%. There was no significant difference among each Binet stages.

CONCLUSIONFISH is a more rapid, accurate and sensitive technique in analysis of chromosome aberrations in CLL. FISH may provide accurate information of molecular cytogenetics for CLL.

Aged ; Aged, 80 and over ; Chromosome Deletion ; Chromosomes, Human, Pair 12 ; Chromosomes, Human, Pair 13 ; Chromosomes, Human, Pair 17 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Male ; Middle Aged ; Trisomy

AIMThe mechanism of vascular endothelial growth factor165 (VEGF165) on intracellular free magnesium ([Mg2+]i) in human umbilical vein endothelial cells (HUVECs) was investigated.

METHODS[Mg2+]i in HUVECs loaded with fluorescent magnesium indicator mag-fura-2 were quantitatively detected the use of intracellular cation measurement system.

RESULTSVEGF165 significantly increased [Mg2+]i in the extracellular Mg2+ and this effect could be blocked by pretreatment with tyrosine kinase inhibitors (tyrphostin A23 and genistein), phosphatidylinositol 3-kinase (PI3K) inhibitors (wortmannin and LY294002) and phospholipase Cgamma (PLCgamma) inhibitor (U73122). In contrast, phospholipase Cgamma (PLCgamma) inhibitor analog (U73343), mitogen-activated protein kinase inhibitors (SB202190 and PD98059) had no effect on the VEGF165-induced [Mg2+]i increase.

CONCLUSIONThe increase of [Mg2+]i by VEGF165 originates from intracellular Mg2+ pool through tyrosine kinase/ PI3K/PLCgamma-dependent signaling pathways.

Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; physiology ; Humans ; Magnesium ; metabolism ; Neovascularization, Physiologic ; drug effects ; Phosphatidylinositol 3-Kinases ; metabolism ; Phospholipase C gamma ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Signal Transduction ; Vascular Endothelial Growth Factor A ; physiology

OBJECTIVETo analyze the global gene expression profile of primary cultured nasopharyngeal carcinoma (NPC) cells using cDNA microarray techniques to screen new candidate genes related to the occurrence and progression of NPC.

METHODSA NPC cell line C666 and primary cultured NPC cells from biopsy specimens in 5 cases were analyzed with microarray techniques in comparison with 3 normal nasopharyngeal epithelial (NPE) biopsy specimens. Several differentially expressed genes identified from the microarray results were verified by fluorescence real-time PCR (FQ-PCR) and immunohistochemistry (IHC).

RESULTSPrimary cultured cells of both NPC and NPE were verified by cytokeratin IHC, EBER1 in situ hybridization and EBV-DNA real-time PCR. Compared with NPE cells, a total of 493 genes in at least 4/6 of the samples were identified to be differentially expressed in the primary cultured NPC cells, including 264 up-regulated and 229 down-regulated ones. Several differentially expressed genes according to the microarray results were confirmed by real-time PCR and IHC.

CONCLUSIONcDNA microarray technique provides an effective and accurate means for global gene expression profiling of primary cultured NPC cells to screen the differentially expressed genes, which may serve as an important basis for studying the mechanism, classification and diagnosis of NPC at the molecular level.

Animals ; Cells, Cultured ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; RNA ; isolation & purification