Cofactor engineering, a vital part of metabolism engineering, changes the redox cofactor regeneration approach. Its main goal is to rebuild the components of metabolic products. The bioconversion of xylose for the production of ethanol is being studied intensively because ethanol is an alternative energy source and a potential liquid fuel. Saccharomyces cerevisiae has been traditionally used in producing ethanol from fermentable sugars but it cannot utilize xylose, only its isomer xylulose. Introduction of the xylose fermentation pathway from Pichia stipitis into S. cerevisiae enables xylose utilization in recombinant S. cerevisiae, but the ethanol yields of xylose fermentation with recombinant S. cerevisiae has been low and large amounts of the byproduct xylitol are produced. The major reason is that the catabolism of xylose with the fungal pathway leads an imbalance of redox cofactor. The process of the catabolism of xylose requires NADPH and NAD~+, both of which have to be regenerated in separated processes. More and more attention has therefore focused on the redox cofactor balance in S. cerevisia. The research progress of cofactor engineering to solve the imbalance of redox cofactor in xylose metabolism recombinant S. cerevisiae was introduced. This included expression of transhydrogenase, increasing the utilization of NADPH, and achieving the anaerobic reoxidation of NADH. Reversing the cofactor specificity of enzymes is another effective way.

Objective Concluding the clinical feature and prognosis of primary testicular lymphoma to improve the understanding of this disease.Methods During 1995 and 2010,17 cases of primary testicular lymphoma treated in Beijing Friendship Hospital of Capital Medical University were retrospectively analyzed of its clinical features,diagnosis,treatment and prognosis.Results Seventeen patients with a mean age of 68 years complained the testicular sohd mass as their first symptoms.The mean tumor diameter was 4.7 cm,and all patients underwent orchidectomy,and testicular non- Hodgkin's lymphoma was confirmed by pathologic examination.Fourteen cases were diffuse large B cell type and 3 cases were anaplastic large cell type.The clinical stage of all the patients was IE.Fourteen cases were followed up (3 cases were lost)with mean follow-up time of 37.8 months by outpatient interview and telephone,all patients were treated with CHOP chemotherapy,and some of them were combined with rituximab and preventive lowdose pelvic radiotherapy.Five cases died of other chronic medical complications,1 case with contralateral testicular metastasis received surgery again.There were 9 tumor free survival cases in total.Conclusion Primary testicular lymphoma is rare and more common in older men.Postoperative pathologic diagnosis is gold standard.The systemic treatment and individual therapy is the first choice for primary testicular lymphoma.

Objective To study the relationship of soluble LAIR (sCD305 and CD3060) expression in recipient serum with cytomegalovirus (CMV) pneumonitis after renal transplantation. Methods Nineteen serum specimens from recipients were divided into CMV pneumonitis group (n=10) and control group (n=9). Then the concentrations of sCD305 and CD3060 were quantitated with sandwich ELISA. The data were analyzed by using student t test. Results sCD305 was skewness distributed in both 2 groups, was 0.000-3.039 μg/L in CMV pneumonitis group and 0.000-8.375 μg/L in con-trol group. CD3060 was skewness distributed in CMV pneumonitis group and the concentration was 0.000-0.017μg/L. CD3060 was mormally distributed in control group and the concentration was 0.046±0.035 μg/L. There was significant difference of CD3060 (P=0.000) concentrations and no sig-nificant difference of sCD305(P=0.316) concentrations in 2 groups, respectively. Conclusions The concentration of CD3060 is low in CMV pneumonitis patients. The combination of CMV PP65 antigen detection and CD3060 detection is helpful for the early and precise diagnosis of CMV pneumonitis in renal transplantation patients.

ObjectiveTo observe protective effects of agmatine (AGM) on inflammatory response and spleen immune function in mice with trauma.Methods Forty-eight adult male C57BL/6 mice were randomly divided into three groups (n= 16 each), including control group, model group (bilateral femoral fracture and removal of 35% of the total blood volume), and AGM group (trauma/hemorrhage & AGM 200 mg/kg). Eight mice in each group were sacrificed at 3 hours and 24 hours, respectively, after modeling, and blood samples and tissue homogenate of spleen and liver were collected. The contents of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-1β) in serum and liver tissue were determined with enzyme linked immunosorbent assay (ELISA). Serum aspartate transaminase (AST), alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer. Spleen proliferation response stimulated with concanavalin A (ConA) was evaluated with methyl thiazolyl tetrazolium colourimetry (MTT).γ-interferon (IFN-γ) and IL-2 releases were determined with ELISA.Results Compared with control group, 3 hours after trauma/hemorrhage, the levels of serum TNF-α, IL-6, and IL-1β in model group were significantly elevated [TNF-α (ng/L): 145.38±31.50 vs. 23.06±11.14, IL-6 (ng/L): 496.94±50.76 vs. 47.13±17.47, IL-1β (ng/L): 321.31±43.02 vs. 29.25±16.24,allP< 0.01]. It was found that AGM treatment could alleviate the increase in serum pro-inflammatory mediators induced by trauma/hemorrhage, such as TNF-α (ng/L:111.56±25.47 vs. 145.38±31.50), IL-6 (ng/L: 412.56±44.33 vs. 496.94±50.76), IL-1β (ng/L: 273.38±45.25 vs. 321.31±43.02,P< 0.05 orP< 0.01). Twenty-four hours after trauma/hemorrhage, serum pro-inflammatory mediators were recovered to the levels in control group. There was no significant difference in TNF-α and IL-6 levels at 3 hours after trauma/hemorrhage among groups. Compared with control group, the expressions of liver TNF-α and IL-6 in model group were increased at 24 hours following trauma [TNF-α (ng/mg): 32.93±4.90 vs. 26.58±2.33, IL-6 (ng/mg): 11.20±1.66 vs. 8.38±0.89,bothP< 0.01]. However, AGM inhibited the level of TNF-α (ng/mg:28.92±3.16 vs. 32.93±4.90) and IL-6 (ng/mg: 9.03±1.28 vs. 11.20±1.66) in the liver as induced by trauma/hemorrhage (P< 0.05 andP< 0.01). At 24 hours after modeling, model group and AGM group had distinctly higher serum AST, ALT, LDH levels than those of control group [AST (U/L): 405.9±31.2, 245.7±22.1 vs. 128.2±15.9; ALT (U/L): 92.1±6.3, 51.6±5.0 vs. 30.1±3.2; LDH (U/L): 606.7±36.3, 478.7±25.3 vs. 384.0±16.6, allP< 0.01]. Nevertheless,the increase in serum AST, ALT and LDH was alleviated in AGM group (allP< 0.01). Meantime, trauma/hemorrhage produced a noticeable depression of proliferation of splenic cells and IFN-γ and IL-2 release stimulated with ConA compared with control group [proliferation rate: (40.97±4.13)% vs. (89.99±7.76)%, IFN-γ(ng/L): 91.6±12.3 vs. 353.2±21.5,IL-2 (ng/L): 53.4±6.4 vs. 91.0±12.2,allP< 0.01]. In contrast, AGM notably restored the capacity of proliferation response of splenic cells [proliferation rate: (74.86±5.75)% vs. (40.97±4.13)%, P< 0.01],enhanced the release of IFN-γ and IL-2 stimulated with ConA [IFN-γ (ng/L): 327.8±23.6 vs. 91.6±12.3, IL-2 (ng/L): 74.8±10.4 vs. 53.4±6.4, bothP< 0.01].Conclusion AGM can dramatically alleviate spleen immunosuppression, excessive inflammation and organ damage induced by trauma/hemorrhage.

OBJECTIVETo observe the effect of parecoxib on morphine dosage in patient-controlled analgesia (PCA) following thoracoscope-assisted thoracotomy.

METHODSA consecutive series of 100 patients undergoing thoracoscope-assisted thoracotomy were randomized into 5 groups and received PCA with morphine doses at 0, 5, 10, 15, and 20 mg given in 200 ml saline (groups P(1), P(2), P(3), P(4), and P(5), respectively). Parecoxib (40 mg) was given in all the patients immediately before the operation, and the mixture (4-5 ml) of lidocaine and ropivacaine was administered into the 3 intercostal spaces upper and lower to the incision before chest closure. PCA was administered for each patient. The visual analogue scale (VAS) at rest and coughing and the respiratory functional parameters were recorded at 1, 2, 4, 8, 12, 24, 36, and 48 h after the start of PCA, and the actual and effective button-pressing times (D(1)/D(2)) in PCA were also recorded.

RESULTSNo patients showed signs of respiratory inhibition within 24 h after the operation, and the resting VAS was comparable between the groups within the initial 6 postoperative hours. At 8 to 24 h postoperatively, the VAS scores at rest and coughing were significantly higher in P(1) group than in the other groups (P<0.05), and no significant differences were found between the groups at 36 to 48 h. D(1)/D(2) in groups P(1) and P(2) were significantly different from those in the other 3 groups at 4-24 h, but no such difference was found between groups P(3), P(4), and P(5).

CONCLUSIONThe application of parecoxib may reduce the dosage of morphine in PCA following thoracoscope-assisted thoracotomy and results in good analgesic effect without affecting the patients respiratory function and sputum elimination.

Adult ; Aged ; Analgesia, Patient-Controlled ; methods ; Combined Modality Therapy ; Double-Blind Method ; Female ; Humans ; Isoxazoles ; administration & dosage ; Male ; Middle Aged ; Morphine ; administration & dosage ; Pain, Postoperative ; drug therapy ; Thoracoscopy ; Thoracotomy ; methods ; Young Adult

OBJECTIVETo determine the expression and possible function of CIC-5 during the rat tooth germ development.

METHODSIsolated total mRNA and protein from the rat tooth germs which were at different stages of development. RT-PCR and Western blot were used to investigate the mRNA and protein expression of the CIC-5 in the rat tooth germ.

RESULTSThe mRNA and protein of CIC-5 expressed in the late bell stage, but undetected inThe mRNA and protein of CIC-5 expressed in the late bell stage, but undetected in the early or middle bell stage.

CONCLUSIONThese results showed the spatial temporal distribution of expression of CIC-5 during the different stages of the rat tooth germ development and suggested that it might contribute to the rat tooth germ development.

Ameloblasts ; Animals ; Odontogenesis ; RNA, Messenger ; Rats ; Tooth Germ

AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.

Objective To explore the clinical applicating and efficacy of free fibula osteomyocutane- ous flap in mandible defect reconstruction in osteoradionecrosis patients.Methods The mandible defects were reconstructed by free fibula flaps with or without muscle cuff.The soft tissue defects were repaired by skin paddles.Status of osteotomy in fibula and flap survival was recorded.The complication in recipient site and donor site,as well as mouth opening and occlusion were reviewed.Facial contour and chewing function after reconstruction were evaluated.Results Patients were followed up 3-16 months.4 free fibula flaps with muscle cuff and 5 without muscle cuff survived well.The size of mandible defects covered from 6cm to 17cm. And the harvested fibula flaps with length of 8.6-17cm were cut into 3 segments in 2 cases,and 2 segments in 5 cases.Fibula flap was divided into 2 segments and overlapped in 2 cases.No serious complication was oh- served in recipient site and donor site.Satisfying esthetic result and normal occlusiong of heath mandible were obtained in all cases.The degree of mouth opening was 2.5-3.3cm.Fair chewing function was revealed in re- constructive region after prosthesia repaired.Conclusion Free fibula osteomyocutaneous flap is relatively ideal reconstruction material of mandible defect in osteoradionecrosis patients for its high survival rate and well esthetic results.

Neuroinflammation is closely associated with the development of neurodegeneration diseases, mental disorders and brain injuries. Innate immunity receptors are the first defense line against infections and injuries, which play a critical role in the neuroinflammation and nervous system diseases.This review,focuses on the type classification,function,regulatory mechanism for the innate immunity receptors,and illustrates their roles and molecular mechanism in the development of neuroin-flammation and nervous system diseases.In addition,we will review the current drugs for treating related nervous system diseases, and the possibility of developing new drugs that target neuroinflammation and using anti-inflammatory drugs clinically.

OBJECTIVETo study the effect of RNA interference (RNAi) targeting E6AP on the proliferation and apoptosis of HeLa cells.

METHODSHeLa cells were cultured and divided into 3 groups: blank control group, cells transfected with nonsense siRNA (small interference RNA), and cells transfected with specific E6AP siRNA. The expressions of E6AP mRNA and protein were detected by RT-PCR and Western blot before and after the transfection respectively. Cell proliferation was determined by methylthiazolyl tetrazolium (MTT). The cell apoptosis index was assessed by flow cytometry.

RESULTSUpon treatment with E6AP siRNA for 24, 48 and 72 h, the expression level of E6AP mRNA decreased 33%, 72% and 70% than siRNA treated group. The protein expression levels in 48 h and 72 h E6AP siRNA groups decreased 38%, 59% comparing with those of the nonsense siRNA treated group (P < 0.05). The proliferative capacity of cells transfectd with E6AP siRNA was significantly lower than blank control group (F = 101.38, P < 0.05) and siRNA treated group (F = 38.64, P < 0.05). The apoptosis index of HeLa cells treated with E6AP siRNA was significantly higher than that of the nonsense siRNA (F = 41.48, P < 0.05) and the blank control group (F = 86.36, P < 0.05).

CONCLUSIONSiRNA targeting can effectively suppress the expression levels of E6AP mRNA, corresponding with a proliferation inhibition and an enhanced apoptosis of HeLa cells.

Cell Line, Tumor ; Cell Proliferation ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; Gene Silencing ; HeLa Cells ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Ubiquitin-Protein Ligases ; antagonists & inhibitors ; metabolism ; Uterine Cervical Neoplasms ; genetics ; metabolism