Acidosis ; etiology ; Humans ; Kidney Diseases ; etiology ; Toluene ; toxicity

Objective To investigate the interference and associated mechanism of hnman tissue kallikrein (HK) gene on renal interstitial fibrosis in rats with 5/6 nephrectomy. Methods Human kallikrein cDNA was packed in a recombinant adeno-associated virus(rAAV)-based plasmid vector. The rAAV-HK was produced by transfection in 293 cells. Twenty-four male Wistsr rats were divided into sham operation and operation groups. The rats with 5/6 nephrectomy were randomly divided into simple operation, control and experiment groups. The rats in experiment group received single dose rAAV-HK via the tail vein with 1×1011 pfu. Before nephrectomy and every month after surgery until the rats were sacrificed, the caudal arterial pressure was measured using tail cuff blood pressure determinator. Three months after HK gene delivery, the rats were sacrificed. The expression of HK in rats was assessed by RT-PCR, Western blot and enzyme-linked immunosorbent assay (ELISA). The pathological changes of renal interstitium were evaluated by Masson stainning, and the distribution of bradykinin B2 receptor (BKB2R) and angiotensin Ⅱ typel receptor (ATIR) was examined by immunohistochemistry. The expressions of BKB2R, AT1R, p-MAPK protein in renal tissue were detected by Western blot. Results Three months after HK gene delivery, the systolic blood pressure of experiment group was significantly decreased compared with the control group [(163±13) nun Hg vs (217±16) mm Hg, P<0.01](1 mm Hg=0.133 kPa). Compared with sham rats, the rats in simple operation group and control group had much more renal interstitial collagen deposition and more serious fibrosis performance, but renal interstitial collagen deposition and fibrosis were significantly ameliorated in the rats of experiment group. In addition, the tubulointerstitial injury index of HK transgenic rats was significantly lower than that of the rats in control group (1.33±0.73 vs 3.01±0.62, P<0.01). Up-regnlating expression of bradykinn B2 receptor protein and down-regulating expression of AT1 receptor and p-MAPK protein were found in renal tissues of experimental group after three months (P<0.05). Conclusion HK gene delivery significantly alleviates renal interstitial fibrosis in rats with 5/6 nephrectomy through regulating the expression of bradykinin B2 receptor, AT1 receptor and p-MAPK in renal tissue.

OBJECTIVETo explore the alterations of apoptosis factor Bcl-2/Bax in the early Parkinson's disease (PD) rats and the protective effect of scorpion venom derived bioactive peptide.

METHODSHealthy male SD rats (180-220 g) were randomly divided into 4 groups (n = 10): early PD model group, sham operation group, scorpion venom derived bioactive peptide control group, scorpion venom derived bioactive peptide therapy group. 6-hydroxydopamine (6-OHDA) was used to prepare the early PD rat model. The immunohistochemistry was used to detect the expression of Bax and Bcl-2 and further explore the mechanism of anti-apoptosis regarding the neuroprotective effect of scorpion venom derived bioactive peptide.

RESULTSThe results indicated that compared with the control rats, the immunostaining of Bax in the brain increased significantly while that of Bcl-2 decreased significantly in the lesion side of 6-OHDA treated rats. Interestingly, scorpion venom derived bioactive peptide could attenuate the above abnormal changes.

CONCLUSIONUp-regulation of Bax and down-regulation of Bcl-2 could participate in the early stage of PD and the anti-apoptotic mechanism could be involved in the neuroprotective effect exerted by scorpion venom derived activity peptide regarding the dopaminergic neuron in the early stage.

Animals ; Apoptosis ; Disease Models, Animal ; Down-Regulation ; Male ; Neuroprotective Agents ; chemistry ; Oxidopamine ; Parkinson Disease ; metabolism ; Peptides ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Scorpion Venoms ; chemistry ; Up-Regulation ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo construct a subtracted cDNA library of differentially expressed genes in eosinophils from asthma patients.

METHODSSuppression subtractive hybridization (SSH) was used to isolate the cDNA fragments of differentially expressed genes in the eosinophils of asthma patients before and after treatment. The cDNA fragments were directly inserted into T/A cloning vector to establish the subtractive library, followed by amplification of the library through E. coli transformation with calcium chloride and screening of blue and white clones of the transformants. One hundred positive bacterial clones were randomly picked and identified by colony PCR.

RESULTSThe amplified library contained more than 3,000 positive bacterial clones. Analysis of the randomly selected 100 white clones by PCR showed that 90% of the clones contained 100-500 bp inserts, which might be the cDNA fragments of differentially expressed genes in eosinophils of asthma patients before treatment.

CONCLUSIONA subtracted cDNA library of differentially expressed genes in the eosinophils of asthma patients before and after treatment is constructed successfully by SSH and T/A cloning techniques, which lays a solid foundation for screening and cloning new specific differentially.expressed genes in the eosinophils of asthma patients.

Asthma ; blood ; genetics ; DNA, Complementary ; genetics ; Eosinophils ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; genetics ; Gene Library ; Humans ; Male ; Middle Aged ; Nucleic Acid Hybridization

The cases of lymphoma accompanied or preceded by Coombs' test positive autoimmune hemolytic anemia (AIHA) have been reported. However, Coombs' test negative AIHA prior to the diagnosis of lymphoma was rarely described. Herein, this article reports a case of non-Hodgkin's lymphoma (NHL) preceded about 1.5 years by Coombs test negative AIHA. A woman aged 69 was diagnosed with HA based on the history and laboratory tests. Further studies revealed that this patient was negative with Coombs' test for IgG, IgM, IgA and C3. After all possible causes of HA, especially malignancies were ruled out, the patient was diagnosed with Coombs' test negative AIHA and treated with prednisolone. The patient responded well initially to steroid treatment. Two recurrences of acute HA were presented at time of 10 months post steroid cessation, and immediately after an attempt to withdraw steroid, respectively, but the hemolysis was effectively controlled by reinstitution of prednisolone. At third recurrence, however, the patient was no longer responding to steroid, and was found with cervical lymphadenopathy. Coombs' test for IgG, IgM, IgA and C3 remained negative. B cell NHL was diagnosed by pathology. After receiving 6 cycles of CHOP chemotherapy, the patient was lymphoma free, but the hemolysis was not improved, however, which was effectively controlled by the following low dose-rituximab (RTX) therapy. The patient was still kept in a remission of lymphoma free of anemia. In conclusion, this report presented a very rare case of NHL with Coombs' test negative AIHA as initial major clinical manifestation.
Aged ; Anemia, Hemolytic, Autoimmune ; diagnosis ; etiology ; therapy ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Coombs Test ; Female ; Humans ; Lymphoma, Non-Hodgkin ; complications ; diagnosis ; Rituximab

Animals ; Dynorphins ; metabolism ; Parkinson Disease ; drug therapy ; metabolism ; Peptides ; pharmacology ; Rats ; Scorpion Venoms ; pharmacology

OBJECTIVETo construct and identification of a lentiviral vector for RNA interference (RNAi) targeting STUB1 gene.

METHODSA pair of complementary small hairpin RNA (shRNA) oligonucleotides targeting STUB1 gene was designed, synthesized and inserted into linearized pMagic 4.0 vector. The recombinant plasmid was identified by double restriction digestion with Age I/EcoR I and DNA sequencing.

RESULTPCR and DNA sequencing showed that the shRNA sequence was successfully inserted into pMagic 4.0 vector. The pMagic 4.0 vector was successfully packaged into lentivirus particles.

CONCLUSIONA lentiviral shRNA expression vector and particles targeting STUB1 gene has been successfully constructed for the further study of the STUB1 gene.

Gene Targeting ; Genetic Vectors ; Lentivirus ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Ubiquitin-Protein Ligases ; genetics

This paper introduces a display module which can be used on multi-mode medical images. The module has a small size and can be easily used for point-selecting puncture diagnosis and treatment, and registration for image fusion control points.
Diagnostic Imaging ; methods ; Image Processing, Computer-Assisted ; methods ; Magnetic Resonance Imaging ; methods ; Software Design ; Tomography, X-Ray Computed ; methods

OBJECTIVETo conduct a meta-analysis on the effects of testosterone on the related factors of metabolic syndrome in hypogonadal males.

METHODSBased on the principles and methods of Cochrane systematic reviews, we searched the PubMed (1980 to August 2009), Embase (1980 to August 2009), the Cochrane Central Register of Controlled Trials and CNKI (1995 to August 2009) , and handsearched some relevant journals and conference proceedings as well. We also identified randomized controlled trials addressing the use of testosterone for the treatment of hypogonadism, screened the retrieved studies according to the predefined inclusion and exclusion criteria, evaluated the quality of the included studies, and performed a meta-analysis on the results of homogeneous studies using the Cochrane Collaboration's RevMan 5.0 software.

RESULTSSix randomized controlled trials were included. The results of analysis indicated that testosterone substitution could significantly ameliorate fasting blood glucose, total cholesterol and insulin resistance in hypogonadism patients, and it could also reduce LDL, HDL, triglyceride and systolic blood pressure, though with no significant difference from the controls. However, there was insufficient evidence to show the effects of testosterone on waist circumference, waist-hip ratio and diastolic blood pressure.

CONCLUSIONExisting clinical evidence has demonstrated the positive effects of testosterone substitution on the improvement of insulin resistance, blood glucose and lipids, but due to the heterogeneity and high risk of bias in the included studies, the evidence might be insufficient to give full support to the demonstration. Further large-scale trials are required to define the metabolic effects of testosterone in the treatment of hypogonadism.

Humans ; Hypogonadism ; complications ; drug therapy ; Male ; Metabolic Syndrome ; complications ; Randomized Controlled Trials as Topic ; Testosterone ; therapeutic use ; Treatment Outcome

OBJECTIVETo investigate the effects of intensive insulin therapy on inflammatory response and prognosis of patients with severe trauma.

METHODSEighty severely injured patients were divided into intensive insulin therapy group (n = 40, IT) and routine therapy group (n = 40, RT) in random pair. At the time of admission, a continuous infusion of insulin (2-4 U/h) was pumped into the patients of IT group to maintain blood glucose level at 6-8 mmol/L. Patients in RT group were given routine treatment without administration of insulin. Fever, organ injury, and mortality of patients in 2 groups were recorded. Venous blood was drawn from patients of 2 groups on the morning of post treatment day (PTD) 1, 3, 5, and 7. Values of TNF-alpha, C-reactive protein (CRP), IL-2, and IL-10 in plasma were assayed.

RESULTSHigh fever appeared in 9 patients in IT group, and WBC exceeded 10.0 x 10(9) for more than 3 days in 17 patients in this group, versus 20 and 29 patients respectively in RT group. Dysfunction of 1 organ appeared in 31 patients in IT group and 30 patients in RT group. Dysfunction of 3 organs appeared in 10 patients in IT group and 19 patients in RT group. Dysfunction of 4 organs appeared in 7 patients in IT group and 12 patients in RT group. In IT group, 4 patients died within 3 post-injury day (PID), and 1 patient died after PID 3 (total case fatality: 12.5%). In RT group, 5 patients died within 3 PID, and 4 patient died after PID 3 (total case fatality: 22.5%). Plasma levels of TNF-alpha and CRP of patients in IT group were significantly lower than those of patients in RT group on PID 3-7 ( P < 0.05 or P < 0.01), while levels of IL-2 and IL-10 of patients in IT group were significantly higher than those of patients in RT group (P < 0.05 or P < 0.01). Plasma levels of TNF-alpha (1.3 +/- 0.6 microg/L) and CRP (55 +/- 16 mg/L) of patients in IT group on PTD 7 were lowered to the trough level, and they were significantly lower than those of patients in RT group (3.0 +/- 0.8 microg/L, 89 +/- 20 mg/L, respectively, P < 0.01).

CONCLUSIONSIntensive insulin therapy can mitigate systemic inflammatory response and improve prognosis of patients with severe trauma.

Adult ; Aged ; Female ; Humans ; Inflammation ; Insulin ; administration & dosage ; Interleukin-2 ; blood ; Male ; Middle Aged ; Prognosis ; Wounds and Injuries ; therapy ; Young Adult