Aerenchyrna formation has been described in depth in a number of species at a histological level. But large gaps remain in our understanding of its regulation as a developmental process. It is attempted to analyse essential mineral elements like K, Mg, Cu, Zn, Ca and P in the cell wall of aerenchyma cells in petioles ofS. trifolia at five different developmental stages by CSEM-EDX technique. At early stage, K and Cl concentrations in cell wall were high up to 36% and 4.3% of dry weight, respectively. It supported the hypotheses that aerenchyma spaces are filled with liquid at early developmental stages of aerenchyma in S. trifolia petiole. Mg concentration was high at stage 2, up to 0.86% of dry weight. Zinc and Cu were detected only at rapid expansion stages, during which the concentrations were up to 1.5% and 2.5%, respectively. Calcium was detected in the cell wall only at mature stages, the concentration was high up to 1.3% of dry weight at stages 4 and 5. These results confirmed that the element concentration of aerenehyma cell wall undergoes dynamic changes during different developmental stages, and a low Ca with high Zn and Cu concentration are needed for cell expansion. Copper and Zn deposition in the cell wall showed a significant positive linear correlation, suggesting that these two elements share same or similar uptake and transport mechanism in plants.

OBJECTIVETo observe the expressions of Calbindin(CB) and Parvalbumin (PV), the two calcium-binding protein, in auditory pathway in mice of wild type C57BL/6J and kit⁺/kitW⁻ ²Bao, a kit gene mutant.

METHODSSix mutated kit gene kit⁺/kitW⁻ ²Bao mice and 6 wild type C57BL/6J (B6) mice were anaesthetized i. p. with chloral hydrate. After the mice were fixed by heart perfusion, the brains were removed and coronal sections were cut with a freezing microtome.

RESULTSWe found that wild type mice had significant expressions of PV on ventral cochlear nucleus, anterior part (AVCN), ventral cochlear nucleus, posterior part (PVCN), inferior colliculus (IC) and auditory cortex (AC). CB was expressed in wild type mice on PVCN and nucleus of the trapezoid body (Tz). The mutant of kit gene induced the less expression of PV on PVCN, IC and AC (P < 0.01), but increased the expression of Tz (P < 0.01). CB could not be observed on PVCN in mutant mice, and the expression of AC was increased( P < 0.01).

CONCLUSIONCB and PV has differential expression level in auditory pathway. Since mutated kit gene can affect expression of PV on PVCN, IC, Tz and AC, as well as CB on PVCN and AC, it suggests that the mutation of kit gene can affect the advanced function of central nervous system in auditory pathway.

Animals ; Auditory Cortex ; metabolism ; Auditory Pathways ; metabolism ; Calbindins ; metabolism ; Inferior Colliculi ; metabolism ; Mice ; Mice, Inbred C57BL ; Mutation ; Parvalbumins ; metabolism ; Pons ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics

AIM: To establish the quality standard for Tongfeng Huadu Tincture (Radix Gentianae Macrophyllae, Radix Angelicae Pubescentis, Rhizoma Chuanxiong, Herba Asari, etc.). METHODS: The Radix Gentianae Macrophyllae, Radix Angelicae Pubescentis, Rhizoma Chuanxiong, Herba Asari of Tongfeng Huadu Tincture were identified by TLC. Strychnine of Tincture was determined by HPLC. RESULTS: The average recovery was 97.07% and RSD and 2.1% (n=5). CONCLUSION: The method is reliable, accurate and specific. It can be used for quality control of Tongfeng Huadu Tincture.

A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
Alkyl and Aryl Transferases ; biosynthesis ; genetics ; Azulenes ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Open Reading Frames ; Polyisoprenyl Phosphates ; Recombinant Proteins ; biosynthesis ; Sesquiterpenes ; metabolism ; Sesquiterpenes, Guaiane ; Thymelaeaceae ; enzymology ; genetics

Objective To evaluate the relationship of clinical symptom to plasmic levels of D-dimer, activated factorⅦ (FⅦa) and tissue factor pathway inhibitor (TFPI)/X a in patients with urticaria. Methods A total of 27 patients with chronic urticaria (CU), 27 patients with acute urticaria (AU) and 26 normal human controls were included in this study. Symptom score was determined and disease course was surveyed in these patients. ELISA was used to detect the plasma levels of D-dimer, FⅦa and (TFPI)/Xa in patients and controls. The relation of clinical symptom and disease course to plasma levels of these parameters was assessed. Results In patients with AU and normal controls, the plasma level of D-dimer was 450.57± 242.13 ng/mL and 266.81±40.68 ng/mL, respectively, the level of FⅦa, 2.23± 0.74 ng/mL and 5.23±1.35 ng/mL, respectively, and the level of TFPI/Xa 0.87±0.13 nmol/L and 0.88 ~ 0.12 nmol/L, respectively. There was a significant difference in the level of both D-dimer and FⅦa (both P < 0.01 ), whereas no differ-ence was observed in that of TFPI/X a (P > 0.05) between patients with AU and normal controls. In addi-tion, increased level of D-dimer and decreased level of FⅦa were noticed in patients with CU compared with those in normal controls (593.80±294.04 ng/mL vs 266.81±40.68 ng/mL, 3.98±0.35 ng/mL vs 5.23± 1.35 ng/mL, both P < 0.01 ), but there was no significant difference in the plasma level of TFPI/Xa (0.87± 0.16 nmol/L vs 0.88±0.12 nmol/L, P > 0.05). Significant difference was observed in the plasma level of D-dimer and FⅦa between patients with AU and CU (450.57±242.13 ng/mL vs 593.80 ±294.04 ng/mL, P < 0.05; 2.23± 0.74 ng/mL vs 3.98± 0.35 ng/mL, P<0.01 ). The plasma level of D-dimer positively corre-lated to the symptom score of patients with CU and those with AU (r= 0.68, P< 0.01; r= 0.82, P< 0.01),but was independent of discase course (P> 0.05). Neither the level of FⅦa nor that of TFPI/Xa correlated to symptom score or disease course of patients (all P > 0.05). Conclusions There is an overactivation of coagulation cascade, consumption of blood coagulation factors and secondary fibrinolysis in patients with urticaria, suggesting that plasma D-dimer and FⅦa may be associated with the clinical symptoms of urticaria.

Objective To investigate the effect of histone deacetylase inhibitor trichostatin A (TSA) on the chemotherapy sensibility of 5-fluorouracil (5-Fu) in colorectal cancer cell line Lovo, and to explore the possible mechanisms.Methods According to the treatment methods, the cells were divided into control group, 5-Fu group, TSA group, TSA preconditioning group and combination group (TSA+5-Fu).MTT assay was used to detect cell proliferation at 24 h, 48 h and 72 h after drugs treatment.Transwell assay was used to test cell invasion after 24 h drugs treatment.Flow cytometer was applied to observe the apoptosis after 24 h drugs treatment.The expressions of thymidylate synthase (TS) were detected by Western blot after 24 h drugs treatment.Results Compared with control group, the 5-Fu group, TSA preconditioning group and combination group had a growth inhibition to Lovo cell at 24 h, 48 h and 72 h (P ＜ 0.05), and compared with 5-Fu group, the growth inhibition of TSA preconditioning group and combination group were distinctive at 48 h and 72 h (P ＜ 0.05).However, the inhibition between TSA preconditioning group and combination group were no significant (P ＞ 0.05).Interfered after 24 h, the number of cells penetrating the matrigel in control group, 5-Fu group, TSA group,TSA preconditioning group and combination group were (25.0±4.2), (16.8±2.8), (19.6± 2.5), (8.2±3.2) and (6.5±2.6), respectively (P ＜ 0.05), and the apoptosis rates were (4.26±1.36) ％, (11.66± 3.18) ％, (8.57±2.69) ％, (39.79±8.53) ％ and (45.18±10.07) ％, respectively (P ＜ 0.05).Compared with control group, the number of cells penetrating the matrigel in the experimental groups was significantly decreased, and the apoptosis rate was significantly increased (P ＜ 0.05).Compared with 5-Fu group, the numbers of cells penetrating the matrigel in TSA preconditioning group and combination group were markedly decreased, and the apoptosis rates were markedly increased (P ＜ 0.05), but the number of cells penetrating the matrigel and the apoptosis rate between TSA preconditioning group and combination group were not different (P ＞ 0.05).The difference of TS expression between control group and 5-Fu group was not significant (P ＞ 0.05).Compared with that in control group and 5-Fu group, TS expressions in TSA group, TSA preconditioning group and combination group were markedly decreased (P ＜ 0.05), but TS expressions among the last three groups were not different (P ＞ 0.05).Conclusion TSA can increase the chemotherapy sensibility of 5-Fu in Lovo cells, which may be dependent on reducing the TS expression.

In recent years (2007 to 2011),although the overall number of rabies cases in China has decreased,there is evidence of emerging or re-emerging cases in regions without previous rabies cases or with low incidence of rabies.To investigate the origin and the factors affecting the spread of rabies in China,specimens were collected from 2007 to 2011 from provinces with emerging and re-emerging cases and tested for the presence of the rabies virus.Positive specimens were combined with sequences from GenBank to perform comparisons of homology and functional sites,and to carry out phylogenetic analyses.Out of these regions,five provinces had 9positive specimens from canine and cattle,and 34 canine or human specimens were obtained from previously high-incidence provinces.Complete sequences of G gene were obtained for these samples.Homology of the sequences of these 43 specimens was 87％-100％ at the nucleotide level and 93.7％ -100％ at the amino acid level.These G gene sequences were combined with reference sequence from GenBank and used to construct a phylogenetic tree.The results showed that 43 specimens were all assigned to China clade I and clade Ⅱ,with all specimens from emerging and re-emerging areas placed within clade I.Specimens isolated from Shanxi and Inner Mongolia in 2011 were distinct from previously-isolated local strains and had closer homology to strains from Hebei,Beijing and Tianjin whereas new isolates from Shanghai were tightly clustered with strains isolated in the 1990s.Finally,Shaanxi isolates were clustered with strains from adjacent Sichuan.Our results suggest that the rabies cases in emerging and re-emerging areas in China in the last 5 years are a consequence of the epidemic spreading from of neighboring provinces and regions experiencing a serious epidemic of rabies.

OBJECTIVETo study the therapeutic effects of posterolateral depression fractures of the tibial plateau through a modified anterolateral approach.

METHODSFrom February 2011 to January 2012,13 patients with posterolateral depression fractures of the tibial plateau were treated through a modified anterolateral approach. There were 8 males and 5 females, ranging in age from 28 to 59 years old (49.2 years old on average). Data from patients were collected retrospectively as follows: X-ray, time of fracture healing and the complications of fracture healing. The patients were evaluated both clinically and radiologically according to the Rasmussen score system.

RESULTSAll the patients were followed up, and the duration ranged from 6 to 18 months (mean 13.7 months). All the patients got bony union. The average radiographic bony union time was 15.1 weeks (ranged, 11 to 17 weeks). No case of secondary articular depression was found. No complications such as malunion or joint stiffness were found. But 1 patient had superficial infection and 1 patient had common peroneal nerve injury. According to the Rasmussen score system,the mean radiological score was 16.50 ± 0.67 (ranged, 13 to 18), and the mean functional score was 25.20 ± 2.21 (ranged, 13 to 30). The mean range of knee motion was (125.3 ± 9.3)° (ranged, 0° to 135°).

CONCLUSIONTreatment of depression fractures of posterolateral tibial plateau with a modified anterolateral approach is a safe method with effective exposure, due to its stable fixation and relatively good outcome with minimal soft-tissue complications. It is regarded as an ideal procedure for depression fractures of posterolateral tibial plateau.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery

OBJECTIVETo explore the clinical outcomes of open reduction and internal fixation with calcaneal locking plates in treating Sanders type II and III calcaneal fractures.

METHODSFrom January 2010 and October 2012, 38 calcaneal fractures with Sanders type II or III were treated with open reduction and internal fixation with calcaneal locking plate. According to the Sanders classification, 15 fractures were classified as type II, 23 fractures as type III. The patients were divided into two groups (group A and B) according to the different fixed methods. Sustentaculum tali was fixed with one screw in group A, including 13 males and 5 females, with a mean age of (38.56±8.03) years old (ranged, 25 to 55). And sustentaculum tali was not fixed in group B, including 16 males and 4 females, with a mean age of (42.35±8.29) years old (ranged, 29 to 53). Clinical effects were evaluated according to the changes of Böhler's angle and the Maryland Foot Score and VAS score.

RESULTSAll patients were followed up from 12 to 20 months with a mean of 14 months. Böhler's angles and subtalar joints obtained satisfactory reconstruction in all patients. One year after operation, the mean Maryland Foot Score was 88.61±7.59 in group A; and was 82.40±9.24 in group B; Maryland Foot Score of group A was higher and foot functional rehabilitation was better than group B. The mean VAS score was 13.39±11.47 in group A; and was 22.50±13.10 in group B; VAS score of group A was lower and foot pain was less than group B.

CONCLUSIONSustentaculum tall screw fixation has advantages of strong fixed strength, high stability, less postoperative pain, rapid functional recovery in treating Sanders type II and III calcaneal fractures.

Adult ; Bone Plates ; Bone Screws ; Calcaneus ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Recovery of Function

Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology