Bilateral Renal Denervation Ameliorates Isoproterenol-Induced Heart Failure through Downregulation of the Brain Renin-Angiotensin System and Inflammation in Rat.

Heart failure (HF) is characterized by cardiac dysfunction along with autonomic unbalance that is associated with increased renin-angiotensin system (RAS) activity and elevated levels of proinflammatory cytokines (PICs). Renal denervation (RD) has been shown to improve cardiac function in HF, but the protective mechanisms remain unclear. The present study tested the hypothesis that RD ameliorates isoproterenol- (ISO-) induced HF through regulation of brain RAS and PICs. Chronic ISO infusion resulted in remarked decrease in blood pressure (BP) and increase in heart rate and cardiac dysfunction, which was accompanied by increased BP variability and decreased baroreflex sensitivity and HR variability. Most of these adverse effects of ISO on cardiac and autonomic function were reversed by RD. Furthermore, ISO upregulated mRNA and protein expressions of several components of the RAS and PICs in the lamina terminalis and hypothalamic paraventricular nucleus, two forebrain nuclei involved in cardiovascular regulations. RD significantly inhibited the upregulation of these genes. Either intracerebroventricular AT1-R antagonist, irbesartan, or TNF-α inhibitor, etanercept, mimicked the beneficial actions of RD in the ISO-induced HF. The results suggest that the RD restores autonomic balance and ameliorates ISO-induced HF and that the downregulated RAS and PICs in the brain contribute to these beneficial effects of RD.

[Construction of Eukaryotic Expression Vector Containing ROP18-ROP12 of Toxoplasma gondii RH Strain].

OBJECTIVE: To construct a recombinant eukaryotic expression plasmid containing ROP18-ROP12 (encoding rhoptry protein 18 and 12) complex gene of Toxoplasma gondii, and examine its expression in eukaryotic cells. METHODS: Recombinant plasmids pVAX1-ROP18 and pVAX1-ROP12 were digested by restriction enzymes BamH I and Xba I . ROP12 gene was cloned into pVAX1-ROP18 to construct the eukaryotic expression plasmid pVAX1-ROP18- ROP12. After colony PCR, enzyme digestion and sequencing, the correct recombinant plasmid pVAX1-ROP18-ROP12 was transfected into HeLa cells. Along with it were groups of empty plasmid, pVAX1-ROP18 and pVAX1-ROP12. Total RNA was extracted from HeLa cells and reverse-transcribed to cDNA. RT-PCR was performed to evaluate mRNA expression of the housekeeping gene ß-actin and ROP18-ROP12 complex gene. Immunofluorescence assay and Western blotting were performed to determine the protein levels of ROP18-ROP12 fusion protein. RESULTS: Colony PCR in recombinant plasmid pVAX1-ROP18-ROP12 showed a specific band at about 2 373 bp, consistent with expectation. The extracted recombinant plasmids were confirmed by Hind III, BamH I and Xba I digestion. Sequencing results showed that the sequence of pVAX1-ROP18-ROP12 was 100% identical to that of T. gondii RH strain ROP18 gene (Accession No. AM075204.1) and 99% identical to that of T. gondii RH strain ROP12 gene (Accession No. DQ096559.1). Further, RT-PCR showed amplification products at 613 bp for ß-actin in all the groups, while only the pVAX1-ROP18-ROP12 transfection group showed amplification products for the ROP18-ROP12 complex at 2,373 bp. In addition, the indirect immunofluorescence assay showed yellow-green fluorescence in HeLa cells transfected with pVAX1-ROP18-ROP12, but not in control cells. Western blotting showed that the ROP18-ROP12 fusion protein was expressed in HeLa cells transfected with recombinant plasmid pVAX1-ROP18-ROP12. CONCLUSIONS: The recombinant eukaryotic plasmid pVAX1-ROP18-ROP-2 is constructed and can be expressed in eukaryotic system.

[Cloning and bioinformatics analysis of rhoptry protein 11 of Toxoplasma gondii].

Total RNA was extracted from tachyzoites of RH strain of Toxoplasma gondii. The open reading frame of ROP11 gene was amplified by using a pair of specific primers designed according to the coding sequence of ROP11 gene (Accession No. DQ077905). The RT-PCR product was digested by restriction enzyme EcoR I and Not I, and then ligated into a pGEX-6P-2 vector. The recombinant plasmid was transferred into E. coli XL-Blue. The positive clones was selected by colony PCR, and confirmed by the double restriction enzyme digestion and sequencing. The RT-PCR product was 1,548 bp. The recombinant plasmid was confirmed by colony PCR and double restriction enzyme digestion. Sequencing results showed that the obtained ROP11 gene was 1 548 bp (Accession No. KC456639). There was a high sequence consistency (99%) between the obtained ROP11 gene sequence and the Toxoplasma ROP11 gene from GenBank. Bioinformatics analysis showed that the ROP11 protein (Mr 57,020) consisted of the signal peptide (amino acids 1-26), 12 conservative domains, a serine/threonine protein kinase catalytic domain (amino acids 170-511), and two potential N-glycosylation sites.

[Endoscopic submucosal enucleation for gastric submucosal tumors originated from muscularis propria layer: clinical analysis of 116 case].

OBJECTIVE: To explore the efficacy and safety of endoscopic submucosal enucleation (ESE) for gastric submucosal tumors (SMTs) originated from muscularis propria. METHODS: A total of 116 patients with gastric SMT originated from muscularis propria underwent ESE in Department of Gastroenterology of the Taizhou Hospital between July 2006 and March 2011. The occurrence of intra-operative and post-operative complications and corresponding treatment were recorded. After the treatment of ESE, the patients were followed up endoscopically. RESULTS: The success rate of operation was 96.6%. The mean time of the procedure was (51.9±16.3) min. Complications included intra-operative bleeding (n=9, 7.8%), perforation (n=20, 17.2%), and post-operative bleeding (n=3, 2.6%). Among them, 5 cases (4.3%) required surgical intervention. None of patient had other complications such as peritoneal abscess or peritonitis. The mean hospitalization time after ESE was 6.1 days. The median follow-up period was 12 months (range, 3-48 months) and there was no residual tumor or recurrence. CONCLUSION: ESE is a safe and feasible treatment for patients with gastric SMT originated from muscularis propria.

Endoscopic submucosal enucleation of small gastric gastrointestinal stromal tumors with cross-shaped incision: report of sixty-nine cases.

BACKGROUND/AIMS: Gastric gastrointestinal stromal tumors (gastric GISTs) are the most common gastric submucosal tumors with potential for malignant transformation. Our aim was to assess the efficacy and safety of ESE for gastric GISTs. METHODOLOGY: Small gastric GISTs were dealt with ESE between May 2007 and October 2010. RESULTS: A total of 69 patients (42 men, 27 women; mean age 47.28±10.10 years) were treated. The mean diameter of the specimens was 1.87±0.57cm (range 0.7-3.0cm). The rates of intra-operative bleeding, delayed bleeding, perforation and surgery related complications were 7.25% (5/69), 1.45% (1/69), 33.33% (23/69) and 5.80% (4/69), respectively. The rate of perforation was 43.2% (19/44) at the fundus of the stomach and 16% (4/25) at the body (p=0.02). The mean time of the procedure was 41.07±10.79 minutes. Nineteen patients with perforation were treated by titanium clips and the rest by laparoscopy. Immunohistochemistry revealed that the positive rates of CD117 and CD34 were 88.41% and 68.12%, respectively. The gastric GISTs were all at low risk. At a mean follow-up period of 17.97±10.75 months (range 1 to 40 months) all of the patients were disease free. CONCLUSIONS: ESE with a cross-shaped incision is possibly a very good choice for small gastric GISTs.

[Cloning, expression and immunoreactivity analysis of rhoptry protein 18 (rop18) from Toxoplasma gondii].

OBJECTIVE: To clone and express rhoptry protein 18 (ROP18) gene of Toxoplasma gondii, and analyze its immunoreactivity. METHODS: The genomic DNA was extracted from T. gondii (RH strain) tachyzoites. TgROP18 gene was amplified by PCR, and cloned into pET30a (+) vector. The constructed pET30a (+)-TgROP18 was transformed into E. coli BL21(DE3) and followed by expression of the protein induced by IPTG. The recombinant protein was analyzed through SDS-PAGE, and identified by Western blotting with mouse anti-T. gondii serum. RESULTS: The TgROP18 gene was about 1 665 bp in length and encoded for a protein of 544 amino acid residues and the former 47 amino acids consisted signal peptide sequences. PCR, enzyme digestion and DNA sequencing confirmed that the recombinant plasmid pET30a (+)-TgROP18 was constructed. Bacteria containing recombinant plasmid pET30a (+)-TgROP18 expressed a soluble protein of His-TgROP18 (M, 59 800) after being induced with IPTG. His-TgROP18 reacted positively with mouse anti-T. gondii serum by Western blotting analysis. CONCLUSION: The soluble His-TgROP18 protein shows certain immunoreactivity.

Intramuscular injection of metoclopramide decreases the gastric transit time and does not increase the complete examination rate of capsule endoscopy: a prospective randomized controlled trial.

BACKGROUND/AIMS: Capsule endoscopy (CE) reaches the cecum in about 80% of cases. Decreasing the gastric transit time (GTT) may increase the complete examination rate (CER). METHODOLOGY: Patients (n=177) were prospectively randomized into 2 groups: the control group (n=88) and the intramuscular injection with metoclopramide (IIM) group (n=89). The OMOM CE system, which has the function of real-time monitoring, was used. The patients were injected with metoclopramide 15 minutes before swallowing the CE in the IIM group. The CE would be sent into the duodenum by gastroscopy if the GTT reached 120 minutes in the two groups. RESULTS: No significant difference was noted between the two groups. Of the 169 cases without gastroscopic help, the mean GTT was shorter in the IIM group (n=87) than the control group (n=82) (p=0.002). But the CER was similar. Of 135 cases without gastroscopic help but reached the cecum, the mean GTT was shorter in the IIM group (n=71) than the control group (n=64) (p=0.015). But the mean small bowel transit time (SBTT) was similar. CONCLUSIONS: Intramuscular injection of metoclopramide decreases the gastric transit time, but it does not change the SBTT or CER of capsule endoscopy in our study.

[Establishment of loop-mediated isothermal amplification (LAMP) for detecting Pneumocystis carinii].

Animal model of Pneumocystis carinii pneumonia (PCP) was established for acquiring lung tissue infected with P. carinii. After DNA from rat lungs was extracted, nuclear ribosome small subunit 18s rDNA of P. carinii was amplified by loop-mediated isothermal amplification method at 63 degrees C for 60 min. The product was digested by restriction enzyme Apal I. The results showed that 18s ribosome DNA (rDNA) of P. carinii was cloned into vector pGEX6p2, and the positive clones screened. Therefore, the loop-mediated isothermal amplification has been established for detecting P. carinii.

[Are parasitic infections advantageous to humans?].

Humans are negatively affected by parasitic infection. However, recent researches revealed that to some extent, parasitic infections are advantageous to humans. Parasitic infections are found to benefit patients of inflammatory bowel disease, diabetes mellitus, autoimmune disease and allergic disorder. Furthermore, they promoted studies on pathogenesis of these diseases, and therefore on safe and effective therapeutic strategy. In addition, by taking the Caenorhabditis elegans as model organism, researchers have made a breakthrough in the area of life science, including signal transduction, functional genomics and drug screening.

[Enzyme change in bronchoalveolar lavage fluid of pneumocystis pneumonia rats and the effect of garlicin treatment].

OBJECTIVE: To study the change of enzymes and effect of garlicin treatment on the change in bronchoalveolar lavage fluid (BALF) of rats with Pneumocystis carinii pneumonia (PCP). METHODS: Wistar rats were injected intramuscularly continually with dexamethasone to establish the rat model of PCP. The experimental rats (group A) were injected intramuscularly with garlicin at a dose of 10 mg/(kg x d) for 5 days in the 3rd, 6th and 9th week respectively, and SMZ/TMP therapy group (B), PCP infected group (C) and normal group (D) were established as controls. Three days after the last treatment, the rats of all groups were killed and BALF was collected without contamination and enzymes AST, ALF, CHE, ALP, LDH, CK, CKMB, HBDH, AFU, 5'NT, ADA were examined. RESULTS: The ALP level in group C [(573.41 +/- 350.63)U/L] was significantly higher than that in group D [(210.56 +/- 114.41) U/L] (q = 4.682, P < 0.01), group A [(392.07 +/- 217.57) U/L] (q = 3.851, P < 0.05), and group B [(325.21 +/- 180.65) U/L] (q = 4.380, P < 0.01); the level of CK, CKMB and 5'NT in group C [948.94 +/- 403.43, 489.47 +/- 254.46 and (6.76 +/- 3.11) U/L respectively] was higher than those in group D [426.22 +/- 319.00, 213.33 +/- 144.54 and (3.22 +/- 1.20) U/L] (q = 4.696, 3.784, 3.812, P< 0.05); there was no significant difference in the level of AST, ALT, CHE, LDH, HBDH, AFU and ADA among the four groups (F = 1.852, 0.958, 2.470, 1.423, 1.178, 1.342, 0.611, P > 0.05). CONCLUSIONS: The level of ALP, CK, CKMB but the ALP level decreases distinctly after the garlicin and 5'NT increases evidently in BALF of PCP infected rats, but the ALP level decreases distinctly after the garlicin treatment.

[Determination of trace elements in hair of mice infected with Trichinella spiralis].

Mice were divided into 3 groups: heavy infection group with 80 mice each was fed with 400 muscle larvae of Trichinella spiralis, light infection group with 60 mice each was fed by 200 larvae, and uninfected control (60 mice) . The content of Cu, Zn and Fe in the dorsal hair samples was measured in the week of 1, 3, 5, 7, 9, 11, 13 and 15 after infection. Results indicated that the content of Zn, Cu and Fe in the two experimental groups reduced considerably in comparison to the control (P < 0.05), especially for that of Zn and Cu. Lower content was found in the heavily infected mice than in those with light infection (P < 0.05).

[Study on the changes of soluble intercellular adhesion molecule-1 level in sera of rats infected with Pneumocystis carinii].

OBJECTIVES: To examine sICAM-1 in Pneumocystis carinii pneumonia (PCP) and the effect of TMP-SMZ therapy on its level and on pathological and immunological changes in rats. METHODS: 50 female Wistar rats were randomly divided into normal group (N group), PCP model group (PCP group) and TMP-SMZ therapy group (SMZ group). 1 mg of dexamethasone was injected intramuscularly twice a week for rats in PCP and SMZ groups to induce PCP. Normal saline was injected for N group in the same way. When the infection was confirmed, TMP-SMZ was given to rats in SMZ group by 25 mg/(kg.d) for 5 days for 3 courses with an interval of 2 weeks. sICAM-1 in serum was detected by ELISA, and the pathological changes in lungs and liver and the Pc in alveoli of lungs were observed. RESULTS: The level of sICAM-1 in PCP and SMZ groups at the 3rd week [(1.847+/-0.50) ng/ml, (1.787+/-0.59) ng/ml] was lower notably than that at 0 week [(2.407+/-0.81) ng/ml, [(2.478+/-0.59) ng/ml respectively] (P<0.05), and then increased gradually. It was significantly higher in PCP group at 9th week [(3.233+/-0.83) ng/ml] and at 12th week [(3.984+/-0.87) ng/ml] than that of 0 week (P<0.05). Its level in SMZ group at 12th week [(3.621+/-l.62) ng/ml] was also higher than that in 0 week [(2.478+/-0.59) ng/ml] (P<0.05). sICAM-1 level in both PCP and SMZ groups at 9th week and 12th week was higher than that of N group (P<0.05, P<0.01). There was no significant difference between SMZ and PCP groups at 9th week and 12th week (P>0.05). CONCLUSION: The sICAM-1 level in rats was low but significantly increases after the induction of PCP.

[Advances in biological taxonomy and classification of human parasites].

Along with the further development of biological science and technology, the taxonomy of organisms as well as classification of parasites have been modified and improved. However the taxonomic system of parasites used in China nowadays was established 25 years ago. This paper outlined the advances of biological taxonomy and introduces the Cox's new classification of parasites so as to promote parasitological research.

[The experimental study of garlicin in treating Pneumocystis carinii pneumonia in rats].

OBJECTIVE: To assess the therapeutic effect of garlicin on Pneumocystis carinii pneumonia (PCP) of immunosuppressed rats. METHODS: The Wistar rats were injected intramuscularly continually with dexamethasone for eight weeks to establish the rat model of PCP. The rats were treated with garlicin, meanwhile control groups without treatment and with SMZ-TMP treatment were established respectively in PCP model. The efficacy was evaluated based on the lung weight, lung/body weight ratio and mean cyst number per 100 fields in lung print smear. RESULTS: The mean lung weight of the rats in garlicin is 1.73 +/- 0.17, lung/body weight ratio is 0.84 +/- 0.12, they are obvious lower than 2.31 +/- 0.35 and 1.25 +/- 0.35 of control (P <0.01). The numbers of cysts in lung print smear is reduced 62.9% compared with control. The results is similar to that with SMZ-TMP. CONCLUSION: Garlicin shows an obvious therapeutic effect on Pneumocystis carinii pneumonia in rats with an efficacy similar to that with SMZ-TMP.