The study aimed at investigation the effect of the transgenic biocontrol bacteria 308R(pCPP430) on the tomato rhizosphere bacterial communities. Strain 308R (Pantoea agglomerans), an adnascent bacterium isolated from tomato, was a biocontrol bacterium. The recombinant cosmid pCPP430 contained the hrp gene cluster of Erwinia amylovora was transformed into 308R Strain. Transgenic biocontrol bacterium 308R(pCPP430) produced Harpin protein which induce hypersensitive response and disease resistance in plants and, surprisingly, increase plant growth. Two complementary methods, sole-carbon-source utilization tests (SCSU) and ERIC-PCR, were used to compare the characterization of bacterial communities from three groups of tomato roots dipped in sterilized water, 308R suspended liquid and 308R(pCPP430) suspended liquid. The cluster analysis and principal components analysis of the number of colonies from SCSU indicated consistent differences in the rhizosphere of each group. Cluster analysis of SCSU data showed that there were consistent differences in the rhizosphere of each group. The rhizosphere bacteria communities of root dipped with 308R(pCPP 430) and 308R have a relative good replication, but the differences of corresponding replicates in dipped with distilled water was increased by degrees. Cluster analysis of ERIC-PCR fingerprints revealed that most of results (eight in ten) have overlaps between roots dipped in water and those dipped in 308R liquid. The methods used in this study may prove a useful approach for the comparison of bacterial communities.

Objective To determine the value and reasonable application of slowly coated vicryl plus and albumin gel in primary suture after common BD exploration. Methods The operation was successfully performed in all 45 patients. The incision of common bile duct was directly sewed up by slowly coated vicryl plus and spurted by albumin gel after the exploration. Results There were no complications such as bile leakage and bile duct stricture etc.The mean payment of hospitalization was low 10.5%. Conclusion This method was safe,feasible and effective.

Objective To discuss the role of activation of nuclear factor ?appa B(NF-?B) in apoptosis of alveolar macrophages(AM) induced by gadolinium chloride(GdCl3) in acute necrotizing pancreatitis(ANP).Methods Thirty sixty adult SD rats were randomized into three groups: normal control group,ANP group and the group treated by GdCl3.ANP was induced by intraductal administration of 5% sodium taurocholate,while the normal control rats received an infusion of normal saline.In GdCl3 treatment group,GdCl3 was injected into dorsal vein of penis right after ANP model was established.AM were harvested by bronchoalveolar lavage six hours after model was established.The generation of TNF-? and IL-1? in bronchoalveolar lavage fluid(BALF),and the level of myeloperoxidase in lung tissue were evaluated.The expression of NF-?B protein in AM was determined by western blot.The apoptosis of AM was detected by agarose gel electrophoresis and flow cytometer.The histological examination of lung tissue was checked.Results The levels of TNF-? and IL-1? in ANP group were significantly higher than the control group and GdCl3 treatment group(P

Background Researches documented that retinal nerve fiber layer thickness (RNFLT) in unaffected carriers of Leber hereditary optic neuropathy (LHON) becomes thickened in different quadrants to different degrees.But the change of their macular thickness is still unclear.Objective This study was to clarify RNFLT and macular thickness by optical coherence tomography (OCT) in unaffected female carriers of LHON families.Methods Five female LHON patients (5 eyes) from 5 LHON families,eighteen unaffected female carriers (18eyes) from 18 LHON families and twenty-five age-matched healthy female controls (25 eyes) were included in this study.The patients and genetic carriers were diagnosed in PLA General Hospital from 2011 September to 2012 October.Regular ocular examination were performed followed by OCT measurement of retinas.The Optic Disc Cube 200×200 and Macular Cube 200×200 protocols were used during the OCT measurement.Average (360°) RNFLT,RNFLT at four quadrantic sections,cube average macular thickness and macular thickness of nine Early Treatment Diabetic Retinopathy Study (ETDRS) sub-areas were compared among the LHON genetic carriers,LHON patients and normal controls.Results Compared to the normal control group,significant reduced values were seen in temporal,superior,nasal and inferior side of sub-area macular thickness in the LHON female carriers (P=0.022,0.046,0.024,0.008).In addition,but no significant differences were found in cube average thickness,central subarea macular thickness,temporal,superior,nasal and inferior side of lateral sub-area macular thickness,average RNFLT,and temporal,superior,nasal and inferior quadrant RNFLT between the LHON female carriers and normal controls (P=0.102,0.051,0.238,0.663,0.1 10,0.104,0.419,0.371,0.158,0.063,0.563).Compared to the unaffected female carrier group,female patients showed significant reductions in cube average macular thickness,temporal,superior,nasal and inferior side of sub-area macular thickness,temporal,superior,nasal and inferior side of lateral sub-area mac ular thickness,average R NFLT and temporal,superior,and inferior quadrant RNFLT (P =0.000,0.000,0.000,0.007,0.002,0.002,0.000,0.000,0.040,0.000,0.016,0.000,0.000) except for the central subarea macular thickness and nasal quadrant RNFLT (P=0.388,0.580).Conclusions Unaffected LHON female carriers show a normal peripapillary RNFLT,but the macular thickness at medial sub-area is thinner.This first report offers an information of macular structure change in unaffected LHON female carriers,which suggest that macular damage appears prior to RNFLT change.

OBJECTIVETo explore the correlation between Pi and Shen by observing the relationship between the metabolism of aristolochic acid (AA) and mRNA and protein expression levels of organic anion transporting polypeptide (oatp) superfamily member 2a1 and 2 b1 (oatp2al and oatp2bl) in renal, small intestinal, and large intestinal tissues of Pi deficiency syndrome (PDS) model rats.

METHODSTotally 46 Sprague-Dawley (SD) rats were randomly divided into four groups, i.e., the blank group (n = 12), the PDS group (n = 22), the AA-I group (n = 6), and the PDS AA-I group (n = 6). PDS model was established by subcutaneously injecting Reserpine at the daily dose of 5 mg/kg for 16 successive days. Carotid intubation was performed in 6 rats selected from the blank group and the PDS group. Pharmacokinetics of AA-I were detected at 5, 15, 30, 45, and 60 min after gastrogavage of AA-I. AA-I concentrations in renal, small intestinal, and large intestinal tissues of 10 rats selected from the PDS group were determined. Normal saline was administered to 6 rats selected from the PDS group and the blank group by gastrogavage. Renal, small intestinal, and large intestinal tissues were collected in the AA-I group and the PDS AA-I group at 60 min after gastrogavage of AA-I. mRNA and protein expression levels of oatp2a1 and oatp2b1 in each tissue were detected using real-time polymerase chain reaction (RT- PCR) and Western blot.

RESULTSCompared with the blank group, plasma concentrations of in vivo AA-I were obviously higher in the PDS group at 15, 30, 45, and 60 min after gastrogavage of AA-I with statistical difference (P < 0.05). Plasma concentrations of AA-I were obviously decreased at 60 min after gastrogavage of AA-I; AA-I concentrations in renal and large intestinal tissues were elevated; AA-I concentrations in small intestinal tissues were obviously reduced in the PDS group. There was no statistical difference in mRNA expression levels of oatp2a1 and oatp2b1 in the aforesaid three tissues of rats between the blank group and the PDS group. Compared with the blank group, mRNA expression levels of oatp2a1 and oatp2b1 decreased in small intestinal tissues of the AA-I group, and the mRNA expression level of oatp2a1 in large intestinal tissues significantly decreased (P < 0.05, P < 0.01). Compared with the PDS group, mRNA expression levels of oatp2a1 and oatp2b1 increased in renal tissues of the PDS AA-I group (P < 0.05); mRNA expression levels of oatp2b1 increased in large intestinal tissues of the PDS AA-I group (P < 0.05).

CONCLUSIONSThe difference in AA-I metabolism might be associated with changed expression levels of oatp2a1 and oatp2b1 in renal, small intestinal, and large intestinal tissues under Pi deficiency induced loss of transportation. Shen and Dachang played important roles in substance metabolism under Pi deficiency state, which proved Pi-Shen correlated in Chinese medical theories.

Animals ; Anions ; Aristolochic Acids ; metabolism ; Drugs, Chinese Herbal ; Kidney ; Medicine, Chinese Traditional ; Organic Cation Transport Proteins ; metabolism ; Peptides ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the numbers and locations of spermatic veins, testicular arteries, and lymphatic vessels in the spermatic cord of the varicocele patient under the laparoscope.

METHODSFifty-seven varicocele patients received laparoscopic ligation of spermatic veins, during which we recorded the numbers and observed the locations of spermatic veins, testicular arteries, and spermatic lymphatic vessels.

RESULTSDuring the surgery, we identified 3.3 ± 1.2 spermatic veins, 1.4 ± 0.9 testicular arteries, and 4.3 ± 1.1 spermatic lymphatic vessels. No statistically significant differences were observed between the two side in the numbers of the spermatic veins, testicular arteries and spermatic lymphatic vessels (P > 0.05). The testicular arteries were seen on the exterior of the spermatic veins and winding around them, while the spermatic lymphatic vessels mostly between the veins.

CONCLUSIONThe spermatic veins, testicular arteries, and lymphatic vessels in the spermatic cord of the varicocele patient have their specific anatomic characteristics. Laparoscopic identification of these vessels may contribute to the surgical treatment of varicocele.

Arteries ; anatomy & histology ; Humans ; Laparoscopy ; Ligation ; Male ; Spermatic Cord ; anatomy & histology ; Testis ; Varicocele ; pathology ; Veins ; anatomy & histology

OBJECTIVETo study the kinetogenic effects of serum containing Semen Arecae, Dandelion, Semen raphani and Atractylodes macrocephala on the colonic smooth muscle cells of rats.

METHODSSerum containing Chinese materia medicas was made according to standard methods. Smooth muscle cells were isolated from the muscle layers of Wistar rat's colon, referred to modified Bitar's method. The contractile response of colonic smooth muscle cells to serum containing Chinese materia medicas (10%, 50%, 100% concentration) and other medicines (blank and 1 x 10(-3) mol/L acetylcholine) were separately observed. The contractility was presented by the decrease of the cell length between the drug groups and the control.

RESULTSSerum containing each Chinese materia medica can make dose-dependent contraction at different concentrations (P < 0.05), but the strongest effect of each serum had no significant difference (P > 0.05).

CONCLUSIONSerum containing Semen Arecae, Dandelion, Semen raphani and Atractylodes macrocephala can make notable contraction on colonic smooth muscle cells in rats.

Animals ; Cells, Cultured ; Colon ; cytology ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Muscle Contraction ; drug effects ; Muscle, Smooth ; cytology ; Myocytes, Smooth Muscle ; drug effects ; Rats

Objective To construct the RNA interference eukaryotic expression vector targeting human centromere protein-I (CENP-I) and to observe its effect on the growth of human embryo kidney 293 cells (HEK 293). Methods The expression vectors of pGenesil-1/CENP-I-siRNA-1. pGenesil-1/CENP-I-siRNA-2 and pGenesil-1/CENP-I-siRNA-3 were constructed by gene recombination and then were transfected into the HEK293 cells by liposome. The expressions of CENP-I at the protein and mRNA levels were detected by Western blotting and fluorescence quality PCR (FQ-PCR). The effective vector and the best transfection time were selected. The growth and the cell cycle of the transfected cells were assessed by MTT assay and flow cytometry. Giemsa was used to stain the transfected cells to calculate the mitotic index. Results Sequence-specific siRNAs targeting CENP-I significantly down-regulated the expression of CENP-I in HEK293 cells. The recombinant plasmid of pGenesil-1/CENP-I-siRNA-3 was the effective vector. After transfecting for 72 h the best inhibited efficiency was achieved. In CENP-I-siRNA transfected cells,the rate of cell growth was decreased markedly. Cells at G 2/M phase and the mitotic index were increased conspicuous compared with the cells transfected with the blank vector or untransfected. Conclusion Down-regulation of CENP-I in HEK293 cells by sequence specific siRNA delays the cell growth and postpones the cell division.

Objective To investigate the protective effect of silencing tumor necrosis factor-α (TNF-α) gene in Kupffer cells on liver ischemic reperfusion injury in rats. Methods We constructed the eukaryotic expression vector of small hairpin RNA targeting rat TNF-α gene. PBS, blank vector or TNF-α shRNA mixture were injected through portal vein 48 h before operation. The animals were divided into 4 groups: sham group, PBS group, blank vector group, and shRNA group. The left lateral and median lobes of the liver were rendered ischemic for 40 min resulting in a segmental (70 %) hepatic ischemia. The serum ALT and AST levels were determined 6 h after reperfusion. The mRNA expression of TNF-α in Kupffer cells, serum TNF-α level, concentration of the malondialdehyde (MDA) and superoxide dismutase(SOD) in the liver tissue were detected. Results Compared with the PBS and blank vector groups, ALT and AST levels were significantly decreased in the shRNA group after reperfusion for 6 h (P＜0. 05), TNF-α mRNA level, serum TNF-α level(56.6 ± 6. 7 pg/ml vs 87.8 ± 8. 7 pg/ml and 96.5±7.3 pg/ml,P＜0. 05), concentration of MDA in the liver tissue(93.4±13.3 nmol/mg vs 133.5±12. 4 nmol/mg and 136.7 ±13. 6 nmol/mg, P＜0.05 )were significantly decreased, and SOD activity was significantly increased(22.4±4.6 U/mg vs 12. 2±3. 1 U/mg and 11. 4±2. 9 U/mg,P＜0. 05). Conclusion RNA interfering TNF-α gene in Kupffer cells can protect the liver against ischemia/reperfusion injury.

Objective:To count and analyze the residue of narcotics for injection after the use in operation rooms in order to find the possibility of risk avoiding and management standardization. Methods:A retrospective research method was adopted to count and analyze 10268 prescriptions including narcotics for injection in operation rooms from June to August in 2014. Results:Ketamine hydro-chloride injection had the largest residue and its remaining amount also occupied the largest proportion of medication. The second one was pethidine hydrochloride injection and the third one was remifentanil hydrochloride for injection. The highest ratio of remaining a-mount in each prescription was ketamine hydrochloride injection, and the second one was morphine hydrochloride injection and pethi-dine hydrochloride injection ranked the third. Conclusion:It is suggested to reduce the specification of medication with the highest ra-tio of remaining amount in each prescription such as ketamine hydrochloride injection, and improve the management efficiency for the use of narcotics in operation rooms.