Optimum harvest time of Tulipa edulis based on comparison of biomass accumulation and medicinal quality evaluation / 中国中药杂志

The optimum harvest time of Tulipa edulis was explored based on biomass accumulation and medicinal quality evaluation. Samples were taken from bud stage (Feb 13th) to dormancy stage (May 14th) and the growth indexes, organs biomasses, drying rate, contents of water-soluble extract and polysaccharides were determined. The results showed that biomass distribution of T. edulis varied with growth center and the bulb gained maximum biomass allocation in the whole growth period. The total biomass accumulation and bulb biomass accumulation　increased in the whole growth period and peaked in fructescence stage. No differences were observed in bulb biomass among fructescence stage, withering stage and dormancy stage. The correlation between bulb biomass allocation and other morphological indexes varied with the harvest time. Bulb dry weight biomass had negative correlation with some morphological indexes of aerial part of T. edulis at bud stage, flower stage and fructescence and had significant positive (P<0.05) or extremely significant positive correlation（P<0.01）with other morphological indexes except for root at bearing fruits stage. The drying rate of bulb of T. edulis increased with the extension of harvest time and peaked in dormancy stage. The water-soluble extract of T. edulis bulb was the highest in pre-growing-stage. The tendency of polysaccharides contents showed a W-shape variation during the harvesting period. The polysaccharides content was the lowest in fructescence stage and was the highest in dormancy stage. Considering the yield and medicinal quality of T. edulis bulb, the optimum harvest time of T. edulis is in the withering stage or early stage of dormancy.

Effects of different drying methods on processing performance and quality in bulbus of Tulipa edulis / 中国中药杂志

Effects of different drying methods including sun drying, steamed, boiled, constant temperature drying (at 40, 50, 60 °C) on appearance, hardness, rehydration ratio, dry rate, moisture, total ash, extractive and polysaccharides contents were studied to provide the basis of standard processing method for Tulipa edulis bulbus. The results showed that the treatments of sun drying and 40 °C drying showed higher rehydration ratios, but lower dry rate, higher hardness, worse color, longer time and obvious distortion and shrinkage in comparison with other drying methods. The treatments of 60 °C constant temperature drying resulted in shorter drying time, lower water and higher polysaccharides content. Drying time is shorter and appearance quality is better in the treatment of steaming and boiling compared with other treatments, but the content of extractive and polysaccharides decreased significantly. The treatments of 50 °C constant temperature drying led to similar appearance quality of bulb to commercial bulb, and it resulted in lowest hardness and highest dry rate as well as higher rehydration ratio, extractive and polysaccharides content, moderate moisture and total ash contents among these treatments. Based on the results obtained, 50 °C constant temperature drying is the better way for the processing of T. edulis bulbus.

Effects of low temperature on dormancy breaking and growth after planting in bulbs of Tulipa edulis / 中国中药杂志

The effect of low temperature storage on dormancy breaking, sprouting and growth after planting of Tulipa edulis was studied. The results showed that starch content and activity of amylases significantly decreased during 10 weeks of cold storage, soluble protein content raised at first then decreased, and the peak appeared at the 6th week. However, total soluble sugar content which in- creased slowly at first than rose sharply and reducing sugar content increased during the storage duration. The bulbs with cold storage treatment rooted in the 6th week, which was about 2 weeks earlier than room temperature storage, but there were less new roots in the late period of storage. After stored at a low temperature, bud lengths were longer than that with room temperature treatment. Cold storage treatment could promote earlier emergence, shorten germination time, prolong growth period and improve the yield of bulb, but rarely affect the emergence rate. It was not beneficial to flowering and fruiting. The results indicated that 6-8 weeks of cold storage was deemed to be the key period of dormancy breaking preliminary.

Effects of flower bud removal and artificial pollination on growth and yield of Tulipa edulis / 中国中药杂志

The study was conducted to explore the response of growth and yield of Tulipa edulis to flower bud removal and artificial pollination. And flower bud removal and artificial pollination were carried out in the squaring period and bloom stage respectively. The morphological index and biomass indicators were determined and the yield was counted in harvest time. Result showed that flower bud removal was beneficial to the growth of T. edulis, resulting in increasing growth index, biomass as well as the yield of bulb. The diameter and dry weight of T. edulis fruit by artificial pollination were increased significantly compared with the control. Seed setting percentage increased to 100%, and the number of seed as well as the single grain weight increased by 69.03% and 16.48%, respectively, which did not significantly affect the bulb production. In conclusion, Flower bud removal treatment accelerates bulb biomass increase, so as to improve its yield. Artificial pollination raised significantly seed setting percentage, seed number as well as the single grain weight.

Correlation between the expression of Ku70 and Ku80 in esophageal squamous cell carcinoma and chemotherapy sensitivity and prognosis / 中国医师进修杂志

Objective To investigate the correlation between the expression of Ku70 and Ku80 in esophageal squamous cell carcinoma and chemotherapy sensitivity and prognosis.Methods The expression levels of Ku70 and Ku80 in esophageal squamous cell carcinoma were measured by immunohistochemistry.The relationship between the expression of Ku70 or Ku80 and clinical pathology parameters,recent curative effect of concurrent chemoradiation and prognosis of the patients were analyzed.Results The positive expression rate of Ku70 and Ku80 was 59.0％ (69/117) and 50.4％ (59/117) in esophageal squamous cell carcinoma.The expression of Ku70 and Ku80 was significandy related to recent curative effect of esophageal squamous cell carcinomas (P =0.017,0.012).The patients with complete response had lower positive expression rate of Ku70 and Ku80 [49.2％(32/65),40.0％(26/65)].But the expression had no relationship with the age,length of lesions in CT,lymph node metastasis of esophageal squamous cell carcinoma (P ＞0.05).There was no relationship between the expression of Ku70 and Ku80 neither (P =0.114).KaplanMeier analysis found Ku70 and Ku80 had worse prognosis in patients with positive expression (Ku70:P =0.004 ; Ku80:P =0.025).In Cox univariate analysis concluded the same result (Ku70:P =0.005,HR =1.971,95％ CI:1.230-3.159; Ku80:P =0.028,HR =1.659,95％ CI:1.057-2.605); but in Cox multivariate analysis,only Ku70 expression was the independent factor (P =0.033).Conclusions The upregulation of Ku70 and Ku80 is correlated to decreased concurrent chemoradiation sensitivity.In addition,the upregulation of Ku70 is correlated to worse prognosis.Ku70 and Ku80 can be a prediction index for predicting concurrent chemoradiation sensitivity in esophageal squamous cell carcinoma.

Effects of advanced glycation end products and its receptor on oxidative stress in diabetic wounds / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro.</p><p><b>METHODS</b>Histological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test.</p><p><b>RESULTS</b>Compared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01].</p><p><b>CONCLUSIONS</b>Abnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.</p>

Relationship between cutaneous glycometabolic disorders and cutaneous neuropathy in diabetic rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To analyze the relationship between cutaneous glycometabolic disorders and cutaneous neuropathy in diabetic rats, and to look for the mechanism of neuropathy and impaired wound healing.</p><p><b>METHODS</b>Eighty male SD rats were randomly divided into the normal control group (NC, n = 20), diabetic group (D, n = 20), aminoguanidine-interfered group (AI, n = 20), and insulin-interfered group (II, n = 20) by drawing lots. Diabetes was reproduced in rats of D, AI, and II groups with intraperitoneal injection of streptozotocin (STZ). Then, rats in AI group were fed with 100 mg×kg(-1)×d(-1) aminoguanidine, while rats in II group were subcutaneously injected with insulin for satisfactory control of blood glucose. Changes in mechanical and heat pain thresholds of pad of hind limb were measured at post injection week (PIW) 2, 4, 8. Skin specimens were collected during PIW 2-8 from pads for determination of contents of glucose, advanced glycation end product (AGE), substance P (SP), calcitonin gene-related peptide (CGRP), and observation of distribution and ultrastructure of skin nerve fibers. Data were processed with t test.</p><p><b>RESULTS</b>The mechanical and heat pain thresholds in D group at PIW 2 [(6.3 ± 1.5) g, (6.0 ± 0.9) s, respectively ] were obviously lower than those in NC group [(13.0 ± 3.2) g, (10.3 ± 1.2) s, with t value respectively 2.71, 3.42, P values all below 0.05]. Contents of glucose and AGE in skin tissue in D group were significantly increased when compared with those in NC group, especially at PIW 8 [(2.85 ± 0.33) mg/g, (31.7 ± 3.2) U/mg of hydroxyproline vs. (0.82 ± 0.22) mg/g, (22.2 ± 1.9) U/mg of hydroxyproline, with t value respectively 1.65, 6.47, P values all below 0.01]. The myelinated nerve fibers were edematous and degenerated, with axons compressed, while the unmyelinated nerve fibers were vacuolated, with microfilament and microtubule disorderly arranged. Content of SP in skin tissue in D group was lower as compared with that in NC group, especially at PIW 2 [(16.8 ± 3.4) pg/g vs. (28.5 ± 5.0) pg/g, t = 2.42, P < 0.01]. There was no obvious difference in content of CGRP between NC and D groups, and also in content of glucose in skin between D and AI groups. Compared with those in D group, content of AGE in AI group at PIW 8 was decreased markedly [(27.2 ± 1.4) U/mg of hydroxyproline, t = 3.38, P < 0.05]; contents of glucose and AGE in II group at PIW 8 were significantly decreased [(1.42 ± 0.38) mg/g, (23.6 ± 1.3) U/mg of hydroxyproline, with t value respectively 1.74, 8.17, P < 0.05 or P < 0.01]. Compared with that in D group, contents of SP in AI and II groups were increased, with a delay in time of trough value. Content of CGRP showed no obvious difference among D, AI, and II groups.</p><p><b>CONCLUSIONS</b>High glucose and accumulation of AGE are key mediators of cutaneous neuropathy and impaired wound healing in diabetes mellitus, which confirms that diabetic wound takes an atypical footing during wound repairing. Aminoguanidine and insulin can reduce contents of glucose and AGE in diabetic skin tissue, and ameliorate diabetic cutaneous neuropathy.</p>

Evolutionary trace analysis of N-myristoyltransferase family / 药学学报

To clarify the important functional residues in the active site of N-myristoyltransferase (NMT), a novel antifungal drug target, and to guide the design of specific inhibitors, multiple sequence alignments were performed on the NMT family and thus evolutionary trace was constructed. The important functional residues in myristoyl CoA binding site, catalytic center and inhibitor binding site of NMT family were identified by ET analysis. The trace residues were mapped onto the active site of CaNMT. Trpl26, Asn175 and Thr211 are highly conserved trace residues and do not interact with current NMT inhibitors, which are potential novel drug binding sites for the novel inhibitor design. Pro338, Leu350, Ile352 and Ala353 are class-specific trace residues, which are important for the optimization of current NMT inhibitors. The trace residues identified by ET analysis are of great importance to study the structure-function relationship and also to guide the design of specific inhibitors.

Construction and characterization of a recombinant fowlpox virus co-expressing F, HN genes of Newcastle disease virus and gB gene of infectious laryngnotracheitis virus / 生物工程学报

The Fusion (F) and Haemagglutinin-Neuraminidase (HN) genes of Newcastle disease virus (NDV) and the glycoprotein B (gB) gene of infectious laryngothracheitis virus (ILTV) as well as a LacZ reporter gene were all inserted into a nonessential gene of fowlpox virus (FPV) 017 strain by homologous recombination. The NDV and ILTV genes were each under the control of a fowlpox virus immediate early/late promoter (LP2EP2) while the LacZ reporter gene expression cassette was regulated by a P11 late promoter. A recombinant FPV harboring the F, HN and gB genes as well as the LacZ gene, designated as rFPV-F/HN/gB/LacZ, was obtained after ten cycles of blue plaque purification. The presence of the NDV and ILTV genes was confirmed by PCR. The expression of the recombinant proteins in rFPV-F/HN/gB/LacZ were characterized by Western blot (F and gB proteins) and indirect immunofluorescence test (F, HN and gB proteins). The results demonstrated that all four foreign proteins, which were encoded within a 10 kb gene fragment, could be expressed authentically and efficiently. Compared to the parental virus, rFPV-F/HN/gB/LacZ showed no obvious difference with respect to virus replication and cytopathogenic effects in chicken embryo fibroblasts (CEF) cell culture. Overall, our work suggests that FPV can be a useful live virus vector for the expression of multi- foreign genes against multiple avian pathogens.

Synthesis and antifungal activity of novel triazole antifungal agents / 药学学报

<p><b>AIM</b>A series of triazole antifungal agents were synthesized to search for novel triazole antifungal agents with more potent activity, less toxicity and broader spectrum.</p><p><b>METHODS</b>Twenty-one 1-(1H-1, 2, 4-triazolyl)-2-(2, 4-diflurophenyl)-3-(4-substituted-1-piperazinyl)-2-propanols were synthesized, on the basis of the three dimensional structure of P450 cytochrome 14alpha-sterol demethylase (CYP51) and their antifungal activities were also evaluated.</p><p><b>RESULTS</b>Results of preliminary biological tests showed that most of title compounds exhibited activity against the eight common pathogenic fungi to some extent and the activities against deep fungi were higher than that against shallow fungi. In general, phenyl and pyridinyl analogues showed higher antifungal activity than that of the phenylacyl analogues.</p><p><b>CONCLUSION</b>Several title compounds showed higher antifungal activities than fluconazole and terbinafine. Compound VIII-1, 4, 5 and IX-3 showed the best antifungal activity with broad antifungal spectrum and were chosen for further study.</p>