The endophytic fungi with plant growth promoting effects were screened by co-culture of each endophytic fungus and seedlings of Dendrobium officinale. Anatomical features of the inoculated roots were studied by paraffin sectioning. Morphological characteristics and rDNA ITS1-5. 8S-ITS2 sequences were applied for the taxonomy of endophytic fungi. The results showed that 8 strains inoculated to D. officinale seedlings greatly enhanced plant height, stem diameter, new roots number and biomass. According to the anatomical features of the inoculated roots, each fungus could infect the velamina of seedlings. The hyphae or pelotons were existed in the exodermis passage cells and cortex cells. The effective fungi could not infect the endodermis and vascular bundle sheath, but which was exception for other fungi with harmful to seedlings. Combined with classic morphologic classification, 2 effective strains were identified which were subjected to Pestalotiopsis and Eurotium. Six species of fungi without conidiophore belonged to Pyrenochaeta, Coprinellus, Pholiota, Alternaria, Helotiales, which were identified by sequencing the PCR-amplified rDNA ITS1-5. 8S-ITS2 regions. The co-culture technology of effective endophytic fungi and plant can apply to cultivate the seedlings of D. officinale. It is feasible to shorten growth cycle of D. officinale and increase the resource of Chinese herbs.
Biomass ; DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; cytology ; growth & development ; microbiology ; Endophytes ; classification ; genetics ; physiology ; Fungi ; classification ; genetics ; physiology ; Host-Pathogen Interactions ; Hyphae ; physiology ; Plant Roots ; cytology ; growth & development ; microbiology ; Plants, Medicinal ; growth & development ; microbiology ; RNA, Ribosomal, 5.8S ; genetics ; Seedlings ; cytology ; growth & development ; microbiology ; Species Specificity

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

Objective: To screen the endophytic fungi with antimicrobial activity from Glehnia littoralis and identify the taxonomy of these fungi. Methods: The endophytic fungi were isolated from G. littoralis by method of tissue isolation. The oxford-cup test was used in screening the endophytic fungi with antimicrobial activity for Escherichia coli, Staphylococcus aureus, and Candida albicans respectively. And the rDNA ITS1-5.8S-ITS2 sequences were applied for the taxonomy of endophytic fungi. Results: Nineteen strains of fungi with antimicrobial activity were attributed to four genera, five families, seven orders, and eight species. In these fungi, there were four strains with antimicrobial activity to E. coli (the max d/D value was 1.07), Fifteen strains with antimicrobial activity to S. aureus (the max d/D value was 0.65), and three strains with antimicrobial activity to C. albicans (the max d/D value was 1.27). Conclusion: Many kinds of fungi with antimicrobial activity inhabit in G. littoralis. This study is helpful to develop the recourses of endophytic fungi and to evaluate the genuineness of G. littoralis.

Objective: To evaluate prospectively the clinical significance of immune thrombocytopenia (ITP) bleeding scale (version 2016 of ITP bleeding scale) recommended in consensus of Chinese experts on diagnosis and treatment of adult primary immune thrombocytopenia (version 2016) by Chinese Medical Association. Methods: A total of 88 patients were assessed by ITP bleeding scale (version 2016) and ITP specific bleeding assessment tool (ITP-BAT) to analyze the association between bleeding score of ITP bleeding scale (version 2016) and platelet counts, gender, disease stage, also to evaluate interinstrument consistency between two bleeding grading systems. Among 47 newly diagnosed ITP patients treated with high dose dexamethasone, bleeding score of ITP bleeding scale (version 2016) and platelet counts were assessed before treatment, one week and two weeks after treatment, respectively, and the responsiveness of ITP bleeding scale (version 2016) was evaluated. Results: Bleeding score of ITP patients was negatively correlated with platelet counts (r=-0.515, P<0.001) . Gender and disease stage had no significant influence on bleeding score (F=4.255, P=0.382; F=5.251, P=0.753) . ITP bleeding scale was in excellent agreement with ITP-BAT. Change of bleeding score of 47 ITP patients before and after high dose dexamethasone treatment had significant difference (z=-5.612, P<0.001) , also, the same result was observed in patients who didn’t get complete response (z=-3.969, P<0.001) . Back-to-back scoring consistency of two doctors was 94.4% (k=0.918) . It took less time to accomplish ITP bleeding scale (version 2016) than that of ITP-BAT [3 (2-6) min vs 7 (4-13) min, z=-8.213, P<0.001]. Conclusion ITP bleeding scale (version 2016) has good responsiveness, strong assessment consistency, close correlation with ITP-BAT and less time-consuming in clinical application. It can be used as an effective tool of condition judgement, risk assessment and efficacy evaluation of ITP patients.

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.
Cloning, Molecular ; DNA, Complementary ; Datura ; enzymology ; genetics ; Escherichia coli ; Hyoscyamine ; chemistry ; Mixed Function Oxygenases ; genetics ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Recombinant Proteins ; genetics ; Scopolamine Hydrobromide ; chemistry

PURPOSE: Acute renal graft rejection can only be definitively diagnosed by renal biopsy. However, biopsies carry a risk of renal transplant injury and loss. Micro-CT is widely used in preclinical studies of small animals. Here, we propose micro-CT could noninvasively monitor and evaluate renal location and function in a mouse kidney transplant model. METHODS: Orthotopic kidney transplantation was performed in a BALB/c -to- C57BL/6j or C57BL/6j-to- C57BL/6j mouse model. After optimizing imaging techniques, five mice were imaged with micro-CT and the findings were verified histologically. RESULTS: Micro-CT can monitor and evaluate renal location and function after orthotopic kidney transplantation. There were no mice deaths while renal transplants were failure. CONCLUSION: We propose that graft micro-CT imaging is a new option that is noninvasive and specific, and can aid in early detection and follow-up of acute renal rejection. This method is potentially useful to improve posttransplant rejection monitoring.
Allografts* ; Animals ; Biopsy ; Follow-Up Studies ; Graft Rejection ; Kidney ; Kidney Transplantation ; Mice* ; Transplants

Objective To investigate the water quality and running status of the dringking water improving project in Xuchang,Pingdingshan and Nanyang,thus to provide basis for scientifically evaluating social effects.Methods Stratified sampling was used in the levels of counties and townships and villages,with the number of checked counties greater than or eaquel to half of the projected counties(13/24),the checked townships more than 40 percent of the projected townships(42/102),the checked villages greater than or eaquel to 30% of the projected villages(56/186).We listened their statement,reviewed the documents,examined carefully the water projects before we made the appraisal.Results More than 95%(129/136)of the projects had water quality reports,among which 90%(122/136)were provided by county level centers for disease prevention and control,75% (102/136) of water samples were collected by centers for disease prevention and control,80%(109/136)ofcounties organized an acceptance check-up group,92%(125/136)projects well preserved water source.Conclusions The dringking water improvement projeets in countryside are basically normal in terms of construction and management,the water supplied is qualified,so the expected goal is achieved.However,duties of each department are not explicit and the communication is inadequate,so collaboration should be reinforced.

OBJECTIVETo determine the expression and possible function of CIC-5 during the rat tooth germ development.

METHODSIsolated total mRNA and protein from the rat tooth germs which were at different stages of development. RT-PCR and Western blot were used to investigate the mRNA and protein expression of the CIC-5 in the rat tooth germ.

RESULTSThe mRNA and protein of CIC-5 expressed in the late bell stage, but undetected inThe mRNA and protein of CIC-5 expressed in the late bell stage, but undetected in the early or middle bell stage.

CONCLUSIONThese results showed the spatial temporal distribution of expression of CIC-5 during the different stages of the rat tooth germ development and suggested that it might contribute to the rat tooth germ development.

Ameloblasts ; Animals ; Odontogenesis ; RNA, Messenger ; Rats ; Tooth Germ

OBJECTIVETo explore the effective therapeutic method in the treatment of idiopathic sudden hearing loss (ISHL).

METHODSOne hundred and eighty-eight cases of ISHL were randomized into a warming-promoting needling group (74 cases), a conventional acupuncture group (56 cases) and a medication group (58 cases). In the conventional acupuncture group, the conventional needling technique was applied to Baihui (GV 20), Fengchi (GB 20), Yifeng (TE 17), Tinggong (SI 19), Touqiaoyin (GB 11) and Zhigou (TE 6) on the affected side. The treatment was given 5 times each week. Totally, the treatment of 6 weeks was required. In the warming-promoting needling group, on the basic treatment as the conventional acupuncture group, the warming-promoting needling technique was applied to Fengchi (GB 20). In the medication group, the intravenous drop with salvia injectio and mecobalamin was prescribed, once per day, for 10 days totally. Meanwhile, Erlong Zuoci Wan was prescribed for oral administration, 8 pills each time, three times a day for 30 days continuously.

RESULTSAll of the three therapeutic methods achieved the effect on ISHL. The total effective rate was 89.2% (66/74) in the warming-promoting needling group, which was better than 62.5% (35/56) in the conventional acupuncture group and 53.4% (31/58) in the medication group (both P<0.01).

CONCLUSIONThe warming-promoting needling techinque achieves the significant efficacy on ISHL. The hearing improvement is superior to that treated with either the conventional needling technique or medication.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Female ; Hearing Loss, Sudden ; therapy ; Humans ; Male ; Middle Aged ; Needles ; Treatment Outcome

To explore an efficient strategy for further development of anticancer fluoroquinolone candidates derived from ciprofloxacin, a heterocyclic ring as the bioisosteric replacement of C3 carboxyl group led to a key intermediate, oxadiazole thiol (5), which was further modified to the bis-oxadiazole methylsulfides (7a-7h) and the corresponding dimethylpiperazinium iodides (8a-8h), respectively. Structures were characterized by elemental analysis and spectra data, and their anticancer activities in vitro against CHO, HL60 and L1210 cancer cells were also evaluated by MTT assay. The preliminary results show that piperazinium compounds (8) possess more potent activity than that of corresponding free bases (7).
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; CHO Cells ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ciprofloxacin ; chemistry ; Cricetinae ; Cricetulus ; Drug Design ; HL-60 Cells ; Humans ; Inhibitory Concentration 50 ; Leukemia L1210 ; Molecular Structure ; Oxadiazoles ; chemical synthesis ; chemistry ; pharmacology ; Piperazines ; chemical synthesis ; chemistry ; pharmacology