Objective To construct the eukaryotic expression plasmid of pEGFPC1uPAR gene and explore the effect on the proliferation and invasion ability of Pam 212 cells. Methods The human uPAR cDNA was cloned by PCR, and inserted into the eukaryotic expression plasmid pEGFPC1. After identification of sequencing, the reconstructive plasmid was transformed transiently into Pam 212 cells, then the cell growth and the invasion ability were evaluated. Results The reconstructive plasmid of pEGFPC1uPAR was validated by sequencing. The reconstructive plasmid can promote the growth of Pam 212 cells and enhance the invasion ability. Conclusion The pEGFPC1uPAR plasmid was constructed successfully and uPAR was confirmed to promote the growth and the invasion ability of Pam 212 cells, which lay the foundation for further studies of uPAR in vivo.

Objective: To clone the MADS-box gene denoted AGL15 from total RNA of Lonicera macranthoides based on previous transcriptome sequencing data and analyze the bioinformatics and expression of the gene. Methods: The gene containing intact open reading frame (ORF) was cloned by reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE). The similarity comparison and homology analysis on the sequence were carried out using bioinformatic method, the coding protein was predicted and the physicochemical properties were analyzed. The expression of the gene in different locations of L. macranthoides was determined by semiquantitative PCR using gene-specific primers. Results: The Lm-AGL15 gene, containing a 795 bp ORF that encoded 264 amino acids, was cloned. The deduced protein sequence had the most similarity to the AGL15 in Vitis vinifera and exhibited two conserved motifs (MADS and K-BOX). Without transmembrane domain, Lm-AGL15 was located in cytoplasm and expressed only in each part. Conclusion: For the first time from L. macranthoides cloning could prolong the period of bud of gene, analysis of the gene expression in different parts of L. macranthoides which would provide a reference for the study of prolonging L. macranthoides bud stage.

Objective To evaluate the nutritional status of vitamin A among urban residents. Methods Using stratified random sampling method,a total of 400 residents from two cities in Zhejiang were selected. They were interviewed by questionnaire,also, the dietary survey, physical measurement and blood sample testing were used. According to Recommended Nutrient Intake( RNI)of vitamin A,to evaluate the intake of Vitamin A and analysis the nutritional status and influenc factors. Results The median of vitamin A concentration was 0. 463( inter-quartile range is 0. 213)μg/mL, and the percentage of severe vitamin A deficiency( VAD ),VAD,insufficiency and adequate serum vitamin A were 0. 25%,0. 50%,6. 75% and 92. 50% respectively. Among 299 individuals who participated in the dietary survey,the median of daily vitamin A intake was 350. 5( inter-quartile range was 351. 5)μgRE. Only 25. 42% of participants' dietary intake of vitamin A met the level of 80% RNI,while 61. 87% of peoples' daily intake of vitamin A under 60%RNI. Male, from the bigger city,older age,high level of triglyceride and more intake of cholesterol may contribute to a higher level of serum vitamin A. Conclusion The majority of urban residents had adequate vitamin A . But the intake of dietary vitamin A still should be paid more attention. Serum vitamin A was associated with age,gender and district.

Background A fatal complication after liver transplantation is anastomotic embolization of the hepatic artery. In order to solve this problem, the portal venous arterialization (PVA) is used to reconstruct the hepatic arterial blood flow. The purpose of this study was to investigate the influence of PVA on rats with acute occlusion of hepatic artery.Methods Rat PVA models were established and then randomly divided into Group 1 (control group), Group 2 (jaundice group), Group 3 (bile duct recanalization group), and Group 4 (portal vein arterilization group). Recanalization of the common bile duct and PVA were performed 5 days after bile duct ligation in the rats. The influence of the PVA on general conditions, hepatic changes of structure and function, portal vein pressure and hepatic micrangium were observed for one month.Results Five days after common bile duct ligation the serum bilirubin, transaminase and alkaline phosphatase levels were significantly increased. Compared with group 1, there was a statistically significant difference (P<0.01). These rats then underwent bile duct recanalization and PVA. After a month, the liver functions and microscopic structures completely returned to normal and, compared with group 1, there was no statistically significant difference in portal vein pressure (P >0.05). Vascular casting samples showed that hepatic sinusoids were slightly thicker and more filled than normal ones and although they had some deformations, the hepatic sinusoids were still distributed around the central vein in radial form.Conclusion Within a month after operation, bile duct recanaiization and PVA do not show obvious adverse effects on liver hemodynamics and hepatic micrangium, and the liver function and microscopic structure can return to normal.

OBJECTIVETo analyze the treatment of 42 patients with acute methanol poisoning because of drinking alcohol containing methanol.

METHODSClinical data of 42 cases of methanol poisoning were collected and analyzed. Methanol concentration in drinking alcohol and blood was determined by gas chromatography (GC). National standard for occupational medicine (GBZ53-2002) was used to diagnose the cases.

RESULTSThe methanol concentration in the alcohol was 16% approximately 46%. 42 Patients (40 males, 2 females), at age of 46.1 (22 approximately 80), took 588.1 ml (50 approximately 2,000 ml) of the alcohol. The average methanol concentration in blood was 1.61 mmol/L (0.03 approximately 23.60 mmol/L). According to clinical diagnosis, there were 17 observed cases, 9 mild acute toxication, and 16 severe acute toxication. Among them, 35 (83.3%) patients were recovered, 2 (4.8%) blind, 4 (9.5%) with neuropsychic sequela and 1 (2.4%) dead after adopting 8 cure measures.

CONCLUSIONTo start using emergency plan for public health events suddenly happened, designate a special treatment hospital, clear blood methanol as soon as possible, correct acidosis, adequately administer folacin and hormone, protect optic nerve and retina, and take comprehensive symptomatic treatment as well as strict monitoring are the keys of clinical cure.

Acute Disease ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Methanol ; poisoning ; Middle Aged ; Poisoning ; therapy ; Public Health Practice

OBJECTIVETo construct a single-chain human anti-EGFR antibody (scFv) and truncated protamine (tP) fusion protein, ScFv/tP, carrying small interfering (si)RNA directed against the heat shock protein Hsp47, a collagen-binding glycoprotein, in order to evaluate the role Hsp47 in apoptosis of hepatic stellate cells.

METHODSA single chain of the human variable fragment was obtained by phage display and fused with the tP gene and with or without (negative control) the Hsp47 siRNA sequences. Following expression and purification of the scFv/tP fusion protein and the scFv/tPHsp47 siRNA fusion protein, internalization capabilities were tested in isolated human hepatic stellate cells and the QSG-7701 human hepatocyte cells with visualization by immunofluorescent staining. The DNA binding ability of the fusion proteins were verified by gel shift assay.Following ScFv/tP-Hsp47 siRNA fusion protein transfection into the human hepatic stellate cells, the levels of Hsp47 mRNA and protein expression were tested by RT-PCR and Western blotting; in addition, effects of siRNA-mediated silencing of Hsp47 on cell proliferation and apoptosis were analyzed by the cell counting kit (CCK)-8, flow cytometry and Western blot detection of the apoptosis marker poly (ADP-ribose) polymerase (PARP).

RESULTSIndirect immunofluorescence revealed that the ScFv/tP fusion proteins were internalized into human hepatic stellate cells but not into the QSG-7701 cells.The ScFv/tP-Hsp47 siRNA fusion protein caused reduced expression of Hsp47 mRNA and protein expression in the human hepatic stellate cells, as well as increased the cells' apoptosis remarkably.

CONCLUSIONThe ScFv/tP fusion protein can be used as a transfection reagent to deliver Hsp47 siRNA into hepatic stellate cells and to mediate apoptosis via blockade of Hsp47 expression.

Apoptosis ; Cell Proliferation ; HSP47 Heat-Shock Proteins ; genetics ; Hepatic Stellate Cells ; cytology ; Humans ; Protamines ; metabolism ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Epidermal Growth Factor ; immunology ; Single-Chain Antibodies ; Transfection

BACKGROUNDCD44v6 plays an important role in invasion and metastasis of tumor, Livin has anti-apoptotic effects. The present study aimed to explore the expression and clinical significance of CD44v6 and Livin in gastric cancer tissue.

METHODSStreptavidin-peroxidase linked immunohistochemical method was used to determine the expression of CD44v6 and Livin in gastric cancer tissue and adjacent normal gastric tissues from 59 patients with histopathologically confirmed gastric cancer, and in gastric tissue specimens of 15 patients with gastric polyps, and 15 patients with chronic non-atrophic gastritis. The chi-square test was used for comparison of the relevant factors, Spearman's rank correlation test was applied for relationship among positive expression of the proteins.

RESULTSThe expresion of CD44v6 was positive in 64.4% of the gastric cancer patients; 5.1%, 0 and 13.3% in specimens of normal tissues adjacent to the cancer tissues, in gastric tissue specimens of patients with gastric polyps, and patients with chronic non-atrophic gastritis, respectively. The expression of Livin was positive in 52.5% of the gastric cancer tissues, 6.8%, 0 and 6.7% in the adjacent normal gastric tissue, specimens of patients with gastric polyps and chronic non-atrophic gastritis, respectively. The expression of CD44v6 was significantly correlated with the depth of invasion, the degree of differentiation, and lymphnode metastasis of gastric cancer (P < 0.05). The positive expression rate of Livin protein was also significantly correlated with degree of differentiation of gastric cancer cells and metastasis to lymphnodes (P < 0.05), but not correlated with the depth of invasion and pathological types (P > 0.05). The expression of CD44v6 and Livin in the gastric cancer tissue was positively correlated (r(s) = 0.286, P = 0.028).

CONCLUSIONSThe increased expression of CD44v6 and Livin in gastric cancer tissue may be closely related with development and progression of gastric cancer. CD44v6 and Livin may be new biological markers of gastric cancer.

Adaptor Proteins, Signal Transducing ; analysis ; physiology ; Aged ; Female ; Humans ; Hyaluronan Receptors ; analysis ; physiology ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; analysis ; physiology ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; physiology ; Stomach Neoplasms ; chemistry ; metabolism ; pathology

OBJECTIVETo evaluate the efficiency of navigation system for orbital wall reconstruction in unilateral orbital fractures.

METHODSFifteen patients (7 male and 8 female) with unilateral orbital fracture underwent orbital reconstruction with the help of intraoperative navigation system. The average age was 34.3 ± 9.5 years. All patients underwent spiral CT scanning preoperatively, and the CT data was imported to the BrainLab navigation system (Germany, BrainLab company). The orbit of the intact side was mirrored to the opposite side as the reference for pre-operative planning. The titanium mesh was mounted on the resin template made by rapid prototyping machine based on the mirrored CT data. When the injury was limited, the hydroxyapatite sheet was used for the orbital wall reconstruction. During the operation, the real-time navigation helped to ensure precise placement. The re-establishing result was assessed based on the postoperative CT data with the following four variables: the volumetric difference between the bilateral orbit, the volume of the herniated soft tissue, the global projection and the discrepancy between the simulated and the achieved position of the reconstructed orbital wall. The reconstructive discrepancy was measured only in the titanium plate grafting cases.

RESULTSThere were no serious complications such as infection, graft rejection and optic nerve injury in any case. Preoperatively, the average degree of enophthalmos was (3.5 ± 1.6) mm, the average volumetric difference between the injured and the unaffected orbit was (4.5 ± 1.8) ml, and the average volume of the herniated orbital soft tissue was (2.1 ± 0.7) ml. Postoperatively, the three values were respectively reduced to (1.3 ± 0.6) mm, (1.8 ± 0.9) ml and (0.7 ± 0.3) ml. The discrepancy of the medial and inferior wall were (2.5 ± 0.6) mm and (2.1 ± 0.4) mm.

CONCLUSIONSThe intraoperative use of navigation system for the orbital wall reconstruction in unilateral orbital fractures can provide reliable accuracy and achieve satisfactory results.

Adult ; Computer Simulation ; Enophthalmos ; diagnostic imaging ; etiology ; surgery ; Female ; Humans ; Male ; Orbit ; diagnostic imaging ; surgery ; Orbital Fractures ; complications ; diagnostic imaging ; surgery ; Reconstructive Surgical Procedures ; methods ; Surgery, Computer-Assisted ; Surgical Mesh ; Titanium ; Tomography, X-Ray Computed

OBJECTIVETo compare the efficacy of computer-navigated surgery and the 3-D skull models and guide plates for the treatment of unilateral delayed zygomatic fractures.

METHODSEleven patients with unilateral delayed zygomatic fractures were treated by computer-navigated surgery (test group) and another 12 patients were treated by 3-D skull models and guide plates as the control group. Quality of reduction was assessed by examination of postoperative axial CT scans through zygomatic arch, the malar prominence and the width of zygomatic arch were measured in both groups.

RESULTSThe difference between bilateral malar prominence was (0.94 ± 0.73) mm in the test group and (1.88 ± 1.82) mm in the control group, there was no significant difference between the two groups (P > 0.05). The difference between bilateral width of zygomatic arch was (0.77 ± 0.51) mm in the test group, less than (3.00 ± 1.81) mm in the control group (P < 0.05). Excessive malar prominence was observed in 6 cases (6/11) in the test group and in 7 cases (7/12) in the control group, whereas malar depression was observed in 5 cases (5/11) in the test group and in 5 cases (5/12) in the control group. Overcorrection of the width of zygomatic arch was observed in 6 cases (6/11) in the test group and in 12 cases (12/12) in the control group, whereas under correction was observed in 5 cases (5/11) in the test group and in 0 cases (0/12) in the control group.

CONCLUSIONSIn the treatment of unilateral delayed zygomatic fractures, symmetry of malar prominence can be achieved by application of computer-navigated surgery or the 3-D skull models and guide plates. Symmetry of the width of zygomatic arch achieved with the computer-navigated surgery is better than that achieved with 3-D skull models and guide plates.

Adult ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Surgery, Computer-Assisted ; Tomography, X-Ray Computed ; Young Adult ; Zygoma ; diagnostic imaging ; surgery ; Zygomatic Fractures ; diagnostic imaging ; surgery

OBJECTIVETo observe the effect of hyperbaric oxygen on the scar formation at the rabbit ears at an early stage.

METHODS16 New Zealand rabbits were used to establish the hypertrophic scar model on the ears, 4 wounds on each ear. The rabbits were randomly divided into hyperbaric group(n = 8) and control group (n = 8). The rabbits in the hyperbaric group received hyperbaric oxygen treatment, inhaling oxygen for 1 hour daily under 2ATA circumstance until the wounds were healed. The wound healing and the scar size, thickness, color and hardness in the ears were recorded. After healing, the scar was taken for histologic study with HE staining, Masson staining and trinitrophenol sirius red staining.

RESULTSIn hyperbaric oxygen group, the healing time of 64 wounds was (16.7 +/- 1.8) d, while it was (20.2 +/- 2.3) d in the control group, suggesting a significant difference between two groups (P < 0.05). The incidence of hyperplastic scar was lower (38/64, 59. 4% ) in hyperbaric oxygen group than that (52/ 64, 81.2%) in control group with significant difference between them (P < 0.05). Compared with the control group, the hyperbaric oxygen group had thicker corium layer and less fibroblasts, well-arranged but rare collagen and less collagen nodus and vortex-like structure under microscope. The hyperplastic index of scar was 3.48 +/- 0.94 in hyperbaric oxygen group and 4.65 +/- 0.76 in control group respectively (P < 0.01). The density of fibroblast in two groups were 186.5 +/- 27.3 (hyperbaric oxygen group) and 246 +/- 41.6 (control group) with statistically significant difference (P < 0.05). Furthermore, the area density of collagen fiber were (31.42 +/- 5.36)% in hyperbaric group and (43.62 +/- 7.36)% in control group (P < 0.05). The amount of collagen I and III was (71.42 +/- 5.36)% and (28.58 +/- 5.36)% in hyperbaric oxygen group, (62.46 +/- 7.32)% and (37.54 +/- 7.32)% in control group (P < 0.05). The ratio of collagen I to III was 2.499 in hyperbaric oxygen group, which was similar to the ratio (4:1) in normal skin, compared with the control group (1.664).

CONCLUSIONSThe hyperbaric oxygen can promote wound healing and effectively inhibit the early hyperplastic scar in rabbits ears.

Animals ; Cicatrix ; Disease Models, Animal ; Ear ; pathology ; Female ; Hyperbaric Oxygenation ; Male ; Rabbits ; Wound Healing