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 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 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

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

Objective To investigate the differences in expression of Mohawk (MKX) transcription factors and collagen of types Ⅰ and Ⅲ in anterior cruciate ligament (ACL) gratis between 2 remodeling outcomes under arthroscopy.Methods Enrolled for this study were 17 patients who had undergone arthroscopic single-bundle ACL reconstruction with autogenous hamstring tendons and secondary arthroscopic exploration 48 to 131 months (average,83.1 months) after removal of tibial internal fixator at Department of Sports Medicine,The First Affiliated Hospital to Shenzhen University from March 2017 to December 2017.They were divided into a good remodeling group (11 cases) and a fair remodeling group (6 cases) according to the graft quality under arthroscopy (synovial and vascular coverage,and apparent tension,thickness and retear of the grafts).During the secondary arthroscopic procedures,biopsy of the central ACL grafts was performed.Moreover,normal ACL tissues were harvested from 8 contemporary controls of ＜ 60 years old who underwent total knee replacement.Immunohistochemical assay and quantificational real-time polymerase chain reaction were conducted to detect the expression of transcription factors and collagen of types Ⅰ and Ⅲ in all the samples.Results In the samples from good remodeling and control groups,there were abundant well-arranged collagen fibers of types Ⅰ and Ⅲ and MKX-positive cells;in the fair remolding group,the collagen fibers of types Ⅰ and Ⅲ and MKX-positive cells were much decreased in number and the fibers were not well arranged.The former 2 groups scored in immunohistochemical assay significantly higher than the latter one (P ＜ 0.05).qRT-PCR showed that the expression levels of MKX gene (0.44 ± 0.30),COL1A1 gene (0.52 ± 0.27) and COL3A1 gene (0.60 ± 0.22) in the fair remolding group were significantly lower than in the control group (1.00 ± 0.00,1.00 ± 0.00 and 1.00 ± 0.00) and than in the good remolding group (0.97 ± 0.67,0.99 ±0.38 and 1.00 ± 0.35) (P ＜ 0.05).Conclusion Good remodeling ACL grafts with histological maturation under arthroscopy are more similar to normal ACL than fair remodeling ACL grafts in expression of MKX transcription factors and collagen of types Ⅰ and Ⅲ.

Objective:To investigate the clinical value of virtual monoenergetic images (VMI) and electron density map (EDM) derived from the dual-layer spectral detector CT (DLCT) in the differential diagnosis of benign and malignant pulmonary ground glass nodules (GGN).Methods:From July 2019 to August 2020, a total of 65 patients with lung GGN (27 benign GGNs and 38 malignant GGNs) confirmed by pathology were retrospectively enrolled in Gulou Clinical Medical College of Nanjing Medical University. All the patients underwent DLCT plain scanning within two weeks before the surgery. The conventional 120 kVp polyenergetic image (PI), EDM and 40-80 keV VMI were reconstructed. The differences of CT and electron density (ED) values between benign and malignant lesions on different images were compared by Mann-Whitney U test. Independent t-test was used to compare the lesion size and χ 2 test was used to analyze the CT features (including lesion location, shape, edge, internal structure, adjacent structure, nodule type) between benign and malignant lesions. Receiver operating characteristic (ROC) curve was used to analyze the efficacy of different energy spectrum quantitative parameters in the differential diagnosis of benign and malignant GGN. The statistically significant CT signs and energy spectrum quantitative parameters were analyzed by logistic regression analysis to find out the independent risk factors of malignant GGN, and then ROC curve analysis was performed for each independent risk factor alone or in combination. Results:There were significant differences in lesion shape, spiculation, lobulation, location and size between benign and malignant groups ( P<0.05). The CT value of pulmonary GGN in PI, 40-80 keV VMI and the ED value in EDM were statistically different between benign and malignant lesions ( P<0.05). The area under ROC curve (AUC) were 0.680, 0.682, 0.683, 0.686, 0.694, 0.676 and 0.722, respectively, among which the ED value had the highest AUC. Binary logistic regression analysis was carried out with GGN shape, spiculation, lobulation, location, size, ED value and CT value in PI, 40-80 keV VMI as independent variables, and malignant GGN as dependent variables. The results showed that ED value (OR=1.045, 95%CI 1.001-1.090, P=0.044), lesion size (OR=1.582, 95%CI 1.159-2.158, P=0.004), spiculation sign (OR=11.352, 95%CI 2.379-54.172, P=0.002) were independent risk factors for malignant GGN. ROC curve analysis showed the AUC of ED value, lesion size, spiculation sign and combination of the three for differential diagnosis of benign and malignant GGN were 0.722, 0.772, 0.698 and 0.885. The AUC for the combined parameters was the largest, with sensitivity of 92.1% and specificity of 74.1%. Conclusion:The diagnostic efficacy of EDM is higher than that of other VMI in the differential diagnosis of pulmonary GGN by DLCT images; The efficacy is further improved when EDM is combined with lesion size and spiculation sign for comprehensive diagnosis.

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 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