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.

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

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 explore the effects of different injection techniques of radiotracer on the visualization rate of internal mammary sentinel lymph nodes (IMSLN) in breast cancer patients.

METHODSA series of 137 consecutive breast cancer patients was included in this prospective study. Fifty-eight patients (group A) received the radiotracer (99)Tc(m)-sulphur colloid injected only into 1-2 points in the breast parenchyma in one quadrant, and seventy-nine patients (group B) received the radiotracer injection into the breast parenchyma in two quadrants of the breast. The differences of IMSLN visualization rates of the two groups were compared and the relevant affecting factors were analyzed.

RESULTSThe IMSLN visualization rate of the group B (70.9%, 56/79) was significantly higher than that of the group A (13.8%, 8/58) (P < 0.001). Both techniques seemed to be reliable to identify sentinel lymph node in the axilla (98.7% vs. 98.3%, P = 0.825). In addition, the visualization rate of internal mammary hotspots (82.2%) was more commonly seen in patients receiving injection of a larger volume of radiotracer ( ≥ 0.5 ml/point) than those receiving a smaller volume of radiotracer (<0.5 ml/point, 55.9%, P = 0.011).

CONCLUSIONSThe modified injection technique (two quadrants, large volume radiotraver, and ultrasound guidance) can significantly improve the visualization rate of IMSLN. Our findings should make the biopsy of IMSLN widely implemented and provide an effective and minimally invasive technique to evaluate the internal mammary lymph node status.

Adult ; Aged ; Axilla ; diagnostic imaging ; pathology ; Breast Neoplasms ; diagnostic imaging ; pathology ; surgery ; Female ; Humans ; Injections ; Lymph Nodes ; diagnostic imaging ; pathology ; Middle Aged ; Prospective Studies ; Radionuclide Imaging ; Radiopharmaceuticals ; administration & dosage ; Sentinel Lymph Node Biopsy ; methods ; Technetium Tc 99m Sulfur Colloid ; administration & dosage

OBJECTIVETo evaluate the short-term clinical results of a new approach of lumbar-pelvic fixation for lumbosacral reconstruction after resection of sacral tumors.

METHODSFifteen patients with sacral tumors underwent lumbar-pelvic fixation using TSRH-3D, CDH-M8 or ISOLA with iliac screws. The lumbosacral stability was evaluated according to the X-ray result to assess the feasibility and therapeutic effect of this approach.

RESULTSX-ray showed that high lumbosacral stability was achieved in all the 15 cases after the operation, and satisfactory therapeutic effect was obtained.

CONCLUSIONLumbar-pelvic fixation with iliac screw is feasible for lumbosacral reconstruction after resection of the sacral tumors, which provides strong internal fixation and produce good clinical outcomes.

Adolescent ; Adult ; Aged ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pelvis ; surgery ; Sacrum ; Spinal Neoplasms ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the expression of beta-endorphin and micro-opioid receptor (MOR) during wound healing process in rat with deep partial-thickness scald.

METHODSThirty-six Wistar rats were randomly divided into control( n = 6, without treatment) , and scald ( n = 30, with 5% TBSA deep-partial thickness scald) groups. Skin specimens from wound were harvested immediately after scald and on 3, 7, 14, 21 post-scald days( PSD) for the determination of 1-endorphin and MOR expression with immunofluorescent staining.

RESULTSbeta-endorphin and MOR were mainly distributed in nerve terminal at the border of dermis and epidermis , keratinocyte in some epidermis , in the fibroblast in dermis , with a weak expression The expression of beta-endorphin peaked in whole layer of skin on 3 PSD( 196 +/-16, P <0. 01) ,while that of MOR was concentrated in keratinocytes in the basal layer and the basement membrane. The expression of MOR was strengthened on 7 PSD with disarrangement of collagen , and it peaked on 14 PSD (306 +/- 23, P < 0.01) with epithelization in some wounds. There was still strong expression of beta-endorphin on 7 and 14 PSD. Complete epithelization was observed in scald group on 21 PSD, with nerve terminal approaching the boundary between the dermis and epidermis, and collagen began to arrange in good order. The expression of P-endorphin in scald group (31 +/-24)was similar to that in control group(30 +/- 18) on 21 PSD, but the expression of MOR (56 +/- 16) was still higher than that in control group (28 +/- 15 ).

CONCLUSIONThe expression of MOR and P-endorphin exhibits chronobiological nature during the process of wound healing,

Animals ; Burns ; metabolism ; Disease Models, Animal ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Opioid, mu ; metabolism ; Wound Healing ; beta-Endorphin ; metabolism

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