Objective:To prepare the polyclonal antibodies against advanced oxidation protein products (AOPP),and to provide an effective agent for research on the pathogenesis of AOPP and assess exactly the relationship between AOPP and relative diseases.Methods:AOPP-rabbit serum albumin (AOPP-RSA) was prepared by treating RSA with hypochloric acid.The rabbit anti-AOPP-RSA polyclonal antibodies were generated and purified by affinity chromatography. The titers and the specificity of the antibodies were measured by ELISA.The plasma AOPP and the localization of AOPP in nephridial tissues of some patients with chronic kidney disease (CKD) were determined using rabbit anti-AOPP-RSA.Results:Titers of the antibodies were 10-6.Purified antibodies reacted specifically with oxidized albumin from different genus,but could not react with normal albumin and glycosylated albumin.The high level of AOPP in plasma from CKD patients was confirmed by Western blot.The antibodies could be used to immunostain AOPP deposition in different regions of kidney tissues from both CKD patients and CKD rat models.Conclusion:We successfully generate rabbit anti-AOPP polyclonal antibodies with high titers and striking specificity.The presence of plasma AOPP and localizations of AOPP in kidney tissues of CKD patients can be demonstrated using the antibodies.The development of anti-AOPP polyclonal antibodies may provide a new tool to explore the pathogensis of AOPP and assess exactly the relationship between AOPP and relative diseases.

Objective:To explore CCR7 expression in splenic dendritic cells and its role in migration and activity of splenic dendritic cells in multiple organ dysfunction syndrome (MODS) in mice.Methods:The MODS model of mice was reproduced by Zymosan injection into peritoneal cavity.The mice were randomly divided into groups of normal,3-6 hours,24-48 hours and 10-12 days post zymosan injection.CD11c and CD205 were analysed by immunohistochemistry;The expression of CD86 and CCR7 of DCs were studied by the flow cytometry analysis.Results:In normal mice,many DC were found at the margin between the red and white pulp.In the 3-6 h and 24-48 h groups,CD86 and CCR7 were strongly up-regulated in the DC,and they distributed in T cells areas.In the 10-12 d group,DC distributed at the margin by the immature form.Conclusion:The CCR7 expression level of DC has close correlations with the migration of DC,CCR7 can be used to evaluate the migration and functional activity of DC in MODS.

Objective To analyze the data of influenza A(H1N1) viruses surveillance and genetic characteristics from Taian City during 2005-2008,so a scientific basis can be provided for the prevention and treatment of influenza.Methods The specimens from Influenza-Like Illness(ILI) were collected.The viruses were isolated with MDCK cell and identified with HAI and RT-PCR.The product of PCR were sequenced.Then the sequences were analyzed through biometric software.Results A total of 121 influenza strains were obtained from 615 specimens,and 4 of them were identified as A(H1N1) subtype.There were 3 strains mutated on several sites.Compared with strains isolated in 2005,there were 5 and 8 mutations in the amino acid sequences of virus strains isolated in 2007 and 2008 respectively.And there were a total of 22 amino acid mutations compared with A/Brisbane/59/2007(H1N1).Conclusions Influenza type A(H1N1) are detected in Taian City.There are several mutations in the amino acid sequences of virus strains isolated in Taian. The antigenic drift of virus strains is due to accumulation of amino acid substitutions

Traditional Chinese medicine is a treasure of Chinese culture, absorbing the wisdom of the Chinese people. Continuous application of new technologies makes traditional Chinese medicine research advance with the times. After several years of development, high-throughput transcriptome study has become a mature research tool in biology. This paper reviewed the advances in medicine transcriptome study, and compared two sequencing platforms, Roche's GS FLX platform and Illumina's HiSeq 2000 platform. Moreover, this paper introduced medicine transcriptome analysis process, with Panax quinquefolius and Lonicera japonica for examples, showing the characteristics of traditional Chinese medicine transcriptome studies. High-throughput transcriptome studies facilitate traditional Chinese medicine research with overall understand of functional genes, give clear elucidation of metabolic pathways, lay molecular foundation for the traditional Chinese medicine research and offer modern interpretation for traditional Chinese medicine theory. However, the current study faces several difficulties, including weak molecular basis, high sequencing cost and staff shortages in data anaysis. In the future, with the development in sequencing technology, the combination of transcriptome and other genomics, such as proteome and metabolome, will lay a solid foundation for the new high-throughput screening and developing model for the traditional Chinese medicine industry.
Biomedical Research ; methods ; trends ; Forecasting ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Plant ; Humans ; Lonicera ; genetics ; Medicine, Chinese Traditional ; methods ; trends ; Panax ; genetics ; Phytotherapy ; methods ; trends ; Transcriptome ; genetics

Through study on the correlation between Atractylodis macrocephala lactones ingredient content and topographic factors, we researched regionalization from topography of five main producing provinces of the country, in order to provide a scientific basis for A. macrocephala reasonable cultivation. By sampling from 40 origins of five main producing provinces of the country, the variation of A. macrocephala lactones ingredient content in different conditions of topographic factors and the effect of altitude, slope and aspect was analyzed by SPSS. Then according to the relationship between A. macrocephala lactones ingredient content and topographic factors, the ecological suitability regionalization was conducted by using ArcGIS based on topographic factors. It is suitable for growth of A. macrocephala in the hilly and mountainous areas of southern whose A. macrocephala lactones ingredient content is in high levels. It is unsuitable for growth of A. macrocephala in Northern plain areas, but we can cultivate A. macrocephala in the hilly and mountainous areas of Northern. The most suitable topographic condition for cultivation of A. macrocephala : altitude 200 meters above, slope 3.00-4.99 degrees.
Altitude ; Atractylodes ; chemistry ; growth & development ; China ; Conservation of Natural Resources ; Drugs, Chinese Herbal ; analysis ; Ecosystem

Objective To report our experience with retroperitoneoscopic nephroureterectomy with excision of a bladder-cuff for renal pelvic and ureteral tumors.Methods Thirty-five patients (21 women and 14 men;mean age,67 years;age range,49 -82 years) with upper urinary tract tumors underwent retro- peritoneal laparoscopic nephrourcterectomy with excision of a bladder-cuff.Of the 35 cases,15 had pelvic tumors and 20 had ureteral tumors;19 cases had the tumors on the right side and 16 on the left.Two cases had ureteral tumors combined with bladder tumors.One case had bilateral ureteral tumors then concomitantly had bladder tumors.The needle electrode was used to circleround incise the bladder thoroughly 0.5 cm away from the ureterostoma.Three trocars in the waist were used for dissecting the kidney;and the ureter was dis- sected as far distally downward.Then an incision of 5-9 cm was created in the lower abdomen to allow dis- section of the distal ureter and bladder-cuff and intact specimen extraction.Results The operation was successful in all 35 patients.The mean operative time was 3.1 h ( range,1.5-6.0h).The mean estimated blood loss was 166 ml (range,20-1600 ml).Four cases received blood transfusion.The patient's activity re- covered in 20-32 h after operation.Postoperative pathology showed transitional cell carcinoma in 30 cases, poorly differentiated adenocareinoma in 2 (ureter),squamous cell carcinoma in 1 (ureter),leiomyosarcoma in 1 (ureter),xanthogranulomatous pyelonephritis in 1.Duodenal leakage occurred in 1 patient who had had dialysis a drainage catheter was placed on the third d after operation,and the patient died of heart failure af- ter 2 months.Postoperative vesical irrigation was performed to prevent tumor recurrence.The mean hospital stay was 11 d.During a mean follow-up of 14 months(range,1-32 months),1 patient developed pelvic me- tastasis and was alive with the tumor.The other 33 patients survived free of tumor to date.No patient had re- current transitional cell carcinoma of the bladder.Conclusions Our data demonstrate that retroperitoneo- scopic nephroureterectomy for renal pelvic and ureteral tumors has shorter incision and more rapid postopera- tive recovery compared with open surgery.Using resectoscope to resect the termination of ureter allows more complete excision of the ureter.

OBJECTIVETo investigate effects of low-dose cyclophosphamide and prednisone (CP) metronomic chemotherapy on microvessel density of bone marrow, serum vascular endothelial growth factor (VEGF) and platelet derived growth factor BB (PDGF-BB)in multiple myeloma (MM) patients.

METHODS54 refractory or relapsed MM patients were treated with CP metronomic chemotherapy consisted of oral cyclophosphamide (CTX, 50 mg/d) and prednisone (Pred, 15 mg/d). Bone marrow and peripheral blood of each patient were collected before and 2, 4, 6 months after treatment. Among the 37 assessable patients, 30 cases were responsive with the response rate of 81.08%. Another 17 cases were follow-uped less than 6 months or failure to obtain serum samples or lost to follow-up. Microvessel density of bone marrow was measured by immunohistochemistry and serum VEGF/PDGF-BB expression was analyzed by ELISA in the 37 assessable patients.

RESULTS2, 4, 6 months following CP metronomic chemotherapy, microvessel densities of bone marrow in the responders were 33.1 ± 4.8/HP, 24.8 ± 3.7/HP, 19.7 ± 2.1/HP respectively; the expressions of VEGF were (394 ± 57) ng/L, (268 ± 32) ng/L and (217 ± 20) ng/L respectively; the expressions of PDGF-BB were (304 ± 31) ng/L, (274 ± 31) ng/L and (196 ± 22) ng/L respectively. After CP metronomic chemotherapy, there were significantly lower of microvessel density, VEGF and PDGF-BB levels than pretreatment \[MVD 48.5 ± 5.9/HP, VEGF (517 ± 60) ng/L, PDGF-BB (484 ± 60) ng/L\]in the responders (P < 0.01). While in the non-responders, after treated by CP metronomic chemotherapy for 2 months, microvessel density, the expression of VEGF and the expression of PDGF-BB were 32.5 ± 4.7/HP, 512 ± 39 ng/L and (452 ± 39) ng/L respectively. There were no significant changes of MVD, VEGF and PDGF-BB levels compared with pretreatment \[MVD 33.2 ± 5.6/HP,VEGF (498 ± 55) ng/L, PDGF-BB (488 ± 44) ng/L\] (P > 0.05).

CONCLUSIONSOur findings suggested that continuous low-dose CP metronomic chemotherapy could decrease microvessel density of bone marrow in MM patients. Furthermore, it down-regulated expression of serum VEGF and PDGF-BB to exert its anti-angiogenesis in MM.

Aged ; Aged, 80 and over ; Angiogenesis Inhibitors ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; administration & dosage ; Female ; Humans ; Male ; Microvessels ; drug effects ; Middle Aged ; Multiple Myeloma ; blood ; blood supply ; drug therapy ; Prednisone ; administration & dosage ; Proto-Oncogene Proteins c-sis ; blood ; Vascular Endothelial Growth Factor A ; blood

The study was aimed to assess the effect of Klotho gene and sinoatrial node pacing channel gene (HCN4 and HCN2) for studying sick sinus syndrome, with Klotho gene under the interference of Plasmid-mediated short hairpin RNA. Twenty-five C57BL/6J mice were divided into four groups, i. e, plasmid shRNA 24h group, plasmid shRNA 12h group, sodium chloride 24h group and sodium chloride 12h group. Plasmid shRNA 50microL (1microg/microL) and sodium chloride 50microl were respectively injected according to mice vena caudalis into those in plasmid shRNA group and sodium chloride group. After 12h or 24h respectively, all mice were executed and their sinoatrial node tissues were cut. The mRNA of Klotho, HCN4 and HCN2 gene were detected by RT-PCR. The results of RT-PCR showed that Klotho, HCN4 and HCN2 mRNA levels were lower compared with those in sodium chloride 12h group after 12h interference interval. The results indicated that there might be the a certain relationship between Klotho gene and sinoatrial node pacing channel gene.
Animals ; Glucuronidase ; genetics ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Plasmids ; genetics ; Potassium Channels ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Sinoatrial Node ; metabolism ; physiology ; physiopathology

Objective: To observe the activation of microglia and the changing rule of inflammatory cytokine as IL-6, IL-10 and nuclear factor-κB (NF-κB) in experimental rabbits after spinal cord ischemia reperfusion (SCIR) injury in order to provide theoretical basis for post-conditioning time. Methods: Rabbit SCIR injury model was established by thoracic aorta balloon occlusion. 54 New Zealand male adult white rabbits were divided into 9 groups: Sham group (the animals received balloon implantation without occlusion), SCIR-0h group (reperfusion was conducted at 0 hour of spinal cord ischemia), SCIR-1h, -2h, -3h, -8h, -24h,-48h and -72h groups. n=6 in each group. The number of normal and apoptosis neurons, the levels of Iba-1, IL-6, IL-10 and NF-κB in spinal tissue were examined and compared among different groups respectively. Results: The number of normal neuron was decreasing with the extended reperfusion time, TUNEL-positive neuron began to increasing in SCIR-8h group and the peak was reached in SCIR-24h group. The expression of Iba-1 began to elevating in SCIR-2h group and the peak was obtained in SCIR-8h group; NF-κB began to rising in SCIR-3h group and the peak was observed in SCIR-8h group; both IL-6 and IL-10 arrived the peak in SCIR-24h group. The expressions of NF-κB, IL-6 and IL-10 were positively related to Iba-1 level. Conclusion: Microglia activation had dynamic changes in experimental SCIR rabbits and the expression levels of NF-κB, IL-6 and IL-10 were positively to microglia activation; post-conditioning time at front and back to microglia activation may reduce neuron injury.

BACKGROUND: Intestinal and lymphoid tissues constitute an important part of intestinal immunity, which plays an important regulatory role in spleen deficiency and hydronephrosis. OBJECTIVE: To observe the effect of acupuncture on T lymphocyte subsets in lymph nodes of rats with spleen deficiency, and to investigate the correlation of spleen deficiency with intestinal immunity and the mechanism of acupuncture for spleen deficiency syndrome. METHODS: Thirty-six female Sprague-Dawley rats were randomly divided into three groups: model, acupuncture and blank control groups. The rat model of spleen deficiency was established by fatigue-induced spleen injury plus abnormal diet for 31 days. Afterwards, the rats in the acupuncture group received acupuncture at Zusanli(ST 36).Urine D-xylose excretion rate was detected during modeling and treatment.Then, the mesenteric lymph nodes were removed, and the changes in T lymphocyte subsets in the mesentericlymph nodes were observed by immunohistochemistry. RESULTS AND CONCLUSION: Urine D-xylose excretion rate under spleen deficiency in the modeling and acupuncture groups was significantly lower than that in the blank control group (P < 0.05 or P < 0.01); after acupuncture, the urine D-xylose excretion rate was significantly increased compared with the modeling group (P < 0.01), but still lower than that in the blank control group (P < 0.05). The count of CD4+T lymphocytes, count of CD8+T lymphocytes and ratio of CD4+/CD8+T lymphocytes were ranked as follows: blank control group >acupuncture group>modeling group(P<0.01 or P<0.05).These results suggest that acupuncture at Zusanli can improve the urine D-xylose excretion rate, regulate the balance of T lymphocyte subsets in mesenteric lymph node of rats with spleen deficiency, thus improving the intestinal immune function, spleen deficiency systems, disorder of intestinal digestive function, intestinal digestion and absorption, as well as anorexia, loose stool, diarrhea and other symptoms of the digestive system.