This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.

BACKGROUND: The nutritional support, as well as the water and electrolyte balance during the perioperative period in the renal transplantation recipients at diuresis stage are important to the functional restoration of transplanted kidneys.OBJECTIVE: To explore the method and opportunity of the nutritional support and the handling of the water and electrolyte balance in perioperative period of renal transplantation.DESIGN, TIME AND SETTING: A retrospective clinical analysis was performed in the Department of Urology, Xijing Hospital from June 2003 to June 2007.PARTICIPANTS: Ninety-six patients of chronic renal failure underwent allograft renal transplantation. They comprised 59 males and 37 females, aged 17-67 years, with a mean of 35.7 years.METHODS: The perioperative physiological features of the renal transplantation recipients were summarized retrospectively. The recipients' condition during the perioperative period was divided into two stages at the opening point of allograft blood current. The vital signs of the patients maintained at a stable level before operation. All patients received blood transfusion since the operation began, and were supplemented with albumin before opening the vessels. Urinary production exceeding 100 mL per hour indicated the beginning of fluid replacement, which was a simplified transfusion for the patients at diuresis stage following renal transplantation.MAIN OUTCOME MEASURES: Blood inosine, urea nitrogen, electrolyte, blood sugar and urine of the patients were detected at one day postoperatively.RESULTS: During 12-16 hours postoperatively, the urinary production was 260-1 200 mL, average 520 mL per hour. Blood routine test showed 8 cases developed mild hyponatremia, accounting for 8.3%, 3 cases occurred high potassium and healed after renal functional recovery, 1 case presented low potassium and healed with supplement therapy. There were no abnormal changes of blood chlorine. The blood glucose among 21 cases (21.9%) was higher than the normal level, and recovered following hormone maneuver. The electrolytes and blood glucose were detected to be normal in other patients, without any case with low calcium or magnesium. The urine specific gravity arranged during 1.010-1.015.CONCLUSION: The colloid such as erythrocytes, blood plasma and albumin should be mainly infused before the opening of allograft blood current. And the water and electrolytes is recommended to administrate promptly and regularly during the diuresis stage. The healing of the stoma benefits from the adequate nutritional support. The metabolic acidosis still should be prevented when the urinary production returns normal.

BACKGROUND:The laparoscopy is superior to open surgery for being less invasive, inducing mild stress reaction and allowing quick recovery after operation, however the effects of laparoscopy on perioperative serum cytokine levels are controversial, and only a few studies discuss these effects among pediatric patients.OBJECTIVE: To compare the changes in perioperative cytokine levels and their clinical significance in pediatric patients undergoing laparoscopy and open surgery.DESrGN: Non-randomized concurrent controlled observation.SETTTNG: Department of Urology in Xijing Hospital of the Fourth Military Medical University of Chinese PLA.PARTICT PANTS: From May 2004 to December 2006, 135 pediatric patients for elective operation were recruited from Department of Urology in Xijing Hospital of the Fourth Military Medical University of Chinese PLA. Sixty-five patients were scheduled for laparoscopic surgery while the remaining 70 patients for open surgery.METHODS: Serum levels of interleukin-1 beta (IL-1β), IL-6, IL-10 and tumor necrosis factor-alpha (TNF-α) were measured at 24 hours before operation, and 3, 24, 48 hours after operation respectively. Duration of hospitalization time of all the children was also recorded.MAIN OUTCOME MEASURES: Levels of IL-1β, IL-6, IL-10 and TNF-α of all the patients were measured 24 hours preoperatively, and 3, 24, 48 hours postoperatively.RESULTS: All the 135 cases were included for statistical analysis. ①There were no significant perioperative changes in cytokine levels after laparoscopic surgery (P ＞ 0.05). In the open surgery group, IL-1β and IL-6 levels increased significantly at 3 and 24 hours after operation (P ＜ 0.05), and normalized within 48 hours postoperatively. No significant perioperative differences were found in IL-10 and TNF-α levels (P ＞ 0.05). The levels of IL-1β and IL-6 were significantly higher in the open surgery group than in the laparoscopic surgery group (P ＜ 0.05). ②Duration of hospitalization was shorter in the laparoscopic surgery group than in the open surgery group [(3.5±1.0), (7.5+1.5) days, P＜ 0.05].CONCLUSTON: Pediatric patients undergoing laparoscopic surgery had less perioperative changes in cytokine levels and quicker recovery.

This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; biosynthesis ; metabolism ; Gene Expression Regulation, Plant ; Open Reading Frames ; Oxidoreductases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Roots ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism ; Protein Structure, Secondary ; RNA, Messenger ; metabolism ; Salvia miltiorrhiza ; genetics ; metabolism ; Sequence Alignment ; Silver Nitrate ; pharmacology ; Synthetic Biology

Objective: To investigate effect of the contact surface between the bridge and the adjacent teeth on the stress distribution of the implant and bone tissue and the displacement of the prosthesis in the cantilever fixed implant bridge restoring missing mandibular central incisors. Methods: Two-dimensional images of the mandible and dentition in healthy adults were obtained using CT data. A three-dimensional finite element model of cantilever fixed bridge supported by implants with mandibular central incisor was established by computer reconstruction technique.The contact surface between the bridge and the adjacent natural tooth was designed as "oval" and "trapezoid". The "trapezoid" has a slightly smaller median diameter on the labial side and a slightly larger medial diameter on the lingual side. Loading of 120 N was applied on the tangential margin of the middle line of the long axis of the bridge 41. The direction was set at 0°, which was parallel to the long axis of the tooth and downward. The buccal to lingual and downward angles were 30°, 45° and 60°, respectively, perpendicular to the long axis of the tooth and 90° to the lingual side.The stress distribution of the implant and surrounding bone tissue and the displacement of the prosthesis were compared between the two models. Results: Under axial and buccolingual loading, the maximum equivalent stress peak in the implant and surrounding bone tissue in the cantilever with trapezoidal contact surface design and the maximum displacement of the prosthesis were lower. Moreover, the distribution of stress was more balanced and the concentration range of stress was smaller. With the loading angle increasing, this trend was more obvious. When loading angle increased to 90°, the maximum equivalent stress and the maximum displacement of the elliptic contact surface model implant and surrounding bone tissue were 196 and 101 MPa and 0.196 mm, respectively, while the trapezoidal contact surface model were 157 and 72 MPa and 0.164 mm, respectively. Conclusions The trapezoidal contact surface of the bridge and the adjacent teeth in the cantilever fixed bridge supported by implants with mandibular central incisor is beneficial to reduce the impact of the leverage on the implant and surrounding bone tissue.

Coronaviruses are a large family of viruses which include viruses that cause the common cold and severe acute respiratory syndrome (SARS) in humans and other diseases in animals. There are considerable genetic diversities within coronaviruses due to their wide rang hosts and their special gene replication and transcription mechanisms. During this process, gene recombinations often occur, resulting in novel subtype or coronavirus emerge constantly. Of note are SARS-like-CoVs and novel HCoV-EMC identified in 2012. This minireview summarized major advances of recently identified coronaviruses, focusing on the genome structures and interspecies jumping mechanism of coronavirus.
Animals ; Coronavirus ; classification ; genetics ; isolation & purification ; Coronavirus Infections ; transmission ; Humans ; Phylogeny

OBJECTIVETo look for the active fraction of ethanol extract of Genkwa Flos (EGF) induced hepatotoxicity and develop an UPLC fingerprint of the active fraction.

METHODTarget fraction of EGF induced hepatotoxicity was guided by the serum biochemical and histopathology methods. The UPLC method was applied to establish the chromatographic fingerprint. The separation was achieved on a BEH C18 column (2.1 mm x 50 mm, 1.7 microm) with a mobile phase consisting of acetonitrile and water containing 0.05% phosphate acid running gradient elution. The detection was carried out at 210 nm and the analysis was finished within 10 min.

RESULTThe chloroform phase of EGF could be responsible for the hepatotoxicity of this herb. The common mode of the UPLC fingerprint was set up under the established condition. There were 17 common peaks in fourteen batches of herbs, eight of which were identified, and the similar degrees of the fourteen batches to the common mode were between 0.890-0.999.

CONCLUSIONIt is easy to locate the chloroform extraction of EGF with hepatotoxicity. And the UPLC fingerprint was developed for the above fraction, which could provide valuable references for safe and effective clinical use of EGF.

Animals ; Asteraceae ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; toxicity ; Flowers ; chemistry ; Humans ; Liver ; drug effects ; Male ; Rats ; Rats, Wistar

A series of new coumarin-based benzotriazole derivatives were successfully synthesized via a multi-step sequence of cyclization, etherification and N-alkylation, and were confirmed by 1H NMR, IR, MS spectra as well as elemental analyses. All these synthesized coumarin compounds were evaluated for in vitro antimicrobial activities against four Gram-positive bacteria, four Gram-negative bacteria and three fungi by two fold serial dilution technique. The bioactive assay showed that all these prepared coumarin benzotriazoles could inhibit the growth of the tested bacterial and fungal strains. Title compounds 11a-11e and 13a-13c were more active than chloromycin on Proteus vulgaris ATCC 6896. Coumarin benzotriazoles 11a and 11b displayed comparable antibacterial efficacy against Staphylococcus aureus ATCC 25923 and Micrococcus luteus ATCC 4698 in comparison with reference drug chloromycin. Compared to fluconazole, compounds 11a-11d displayed stronger inhibition on Aspergillus fumigatus ATCC 96918. Moreover, coumarin-based benzotriazoles in combination with antibacterial chloromycin or antifungal fluconazole, showed notable antimicrobial efficacy with less dosage and broader antimicrobial spectrum. More importantly, fluconazole-insensitive A. fumigatus and methicillin-resistant Staphylococcus aureus N 315 (MRSA) were sensitive to these combined drugs.
Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Antifungal Agents ; chemical synthesis ; chemistry ; pharmacology ; Aspergillus fumigatus ; drug effects ; Chloramphenicol ; pharmacology ; Coumarins ; chemical synthesis ; chemistry ; pharmacology ; Drug Synergism ; Fluconazole ; pharmacology ; Fungi ; drug effects ; Gram-Negative Bacteria ; drug effects ; Gram-Positive Bacteria ; drug effects ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Staphylococcus aureus ; drug effects ; Triazoles ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETo establish a mouse model of hypospadias induced by benzoate estradiol to further the studies on the molecular mechanisms of hypospadias.

METHODSA total of pregnant mice were randomly divided into 5 groups, Group A, B, C, D and E, and injected subcutaneously (sc) with estradiol benzoate at the dose of 0, 0.2, 1, 5 and 25 mg x kg(-1) d(-1) respectively from the 12th to the 16th gestational day. The mortality of the newborn mice was recorded and the male neonates of 2 pregnant mice from each group were anatomized to observe the testis position and prostate agenesis on the delivery day. Examinations were made for urethra and cryptorchidism on the 28th postnatal day.

RESULTSThe death rates of the neonates in Group A, B, C, D and E were 21.6%, 21.5%, 41.4%, 56. 6% and 75.0%, respectively. Hypospadias was detected in Group C (3.3%, 1/30), D (20.0%, 4/20) and E (23.0%, 3/13), with significant difference between Group D and A (P < 0.05) and E and A (P < 0.05), but not between Group D and E (P > 0.05). Cryptorchidism was found in Group C (6.6%, 2/30) , D (30.0%, 6/20) and E (61.6%, 8/13), with significant difference between Group D and A (P < 0.05) and E and A (P < 0.05) , but not between Group D and E (P >0.05).

CONCLUSIONExposure of pregnant mice to large dose of estradiol benzoate can induce hypospadias and cryptorchidism in their neonates. And the right dose of estradiol benzoate for the establishment of the mouse model of hypospadias should be 5 mg x kg(-1) x d(-1).

Animals ; Animals, Newborn ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Estradiol ; analogs & derivatives ; toxicity ; Female ; Hypospadias ; chemically induced ; Male ; Mice ; Mice, Inbred ICR ; Pregnancy ; Random Allocation

Objective To study the effects of ribosomal protein L41 (RPL41) on the proliferation and apoptosis of human retinoblastoma Y79 cells and its underlying mechanisms.Methods Y79 cells were seeded in RPMI 1640 medium containing 10％ fetal bovine serum for passage culture.Then the cells were divided into control group,with cells left untreatment,(40 μmol · L-1,80 μmol · L 1 and 120 μmol · L-1) RPL41 treatment group according to the concentration.Next CellTiter-Glo fluorescence cell viability testing system was used to observe the viability of Y79 cells in all groups,and flow cytometry was applied to measure the cell apoptotic rate in 100 μmol · L 1 RPL41 treatment group,with Hoechst staining for the observation of nuclear morphometry of apoptotic cells,and finally,Western blot was used to determine the expression of activating transcription factor 4 (ATF4) of each group.Results Compared with the control group,the viability of Y79 cells in the 40 μmol · L-1 RPL41 treatment group was (97.9 ± 1.5) ％,with no significant difference (P =0.055);and the viability in the 80 μmol · L-1 and 120 μmol · L-1 RPL41 treatment group was (87.6 ± 1.8)％ and (63.9 ± 2.0) ％,respectively,both of which were significantly different from the control group (both P ＜ 0.05),so RPL41 inhibited the viability of Y79 cells,and 100 μmol · L-1 RPL41 promoted the apoptosis of Y79 cells,with the apoptotic rate of (17.33 ± 2.47)％.Compared with normal cells,the apoptotic cells in the 100 μmol · L 1 RPL41 treatment group showed bright color and smaller cell volume by Hoechst staining.Western blot showed that PRL41 significantly decreased the expression of ATF4 protein and the expression of ATF4 protein in the 40 μmol · L 1,80 μmol · L-1 and 120 μmol · L 1 treatment group were 0.76 ± 0.04,0.29 ± 0.04,0.29 ± 0.05,respectively,all of which were significantly different from the control group (all P ＜ 0.01).Conclusion RPL41 can inhibit the proliferation and promote the apoptosis of human retinoblastoma Y79 cell,and its mechanism may be related to the expression of ATF4.