A total of 152 strains of endophytic fungi were isolated from the barks of Eucommia ulmoides in three regions (Lueyang country, Zunyi country, Cili country). Based on morphological characteristics and analysis of ITS sequences, these strains were identified into 8 genera. Thereinto Phomopsis, Diaporthe and Alternaria were common genera to Eucommia barks from different sites. But the dominant genus was different: Alternaria was the dominant genus in the barks from Cili country, and Phomopsis was the dominant genus from Zunyi country, then Diaporthe was the one from Lueyang country. According to the similarity coefficient, the composition of the endophytic fungi was distinctly different between the barks from three sites. The diversity and species richness in Lueyang country and Cili country were found higher than those in Zunyi country. The evenness of endophytic fungi was 0.936 5 in Lueyang county, which was higher than 0.737 1 or 0.641 0 in Cili county or Zunyi county, respectively. After phylogenic analysis and calculating the genetic distances of typical strains belong to Phomopsis and its perfect stage--Diaporthe, there was very high genetic diversity in the two genera from our study. In conclusion, the community structure and diversity of endophytic fungi were significant different in Eucommia barks from the three habitats.
DNA, Fungal ; genetics ; DNA, Intergenic ; genetics ; Ecosystem ; Endophytes ; classification ; physiology ; Eucommiaceae ; microbiology ; Fungi ; classification ; genetics ; physiology ; Phylogeny ; Plant Bark ; microbiology

Objective: To evaluate the anti-tumor and side effects of activated-HLA haploidentical peripheral blood tem cells (haplo PBSCs) in the treatment of advanced refractory solid tumor patients. Methods: Forty-two patients with advanced refractory tumor, who were diagnosed in our hospital from Oct. 2004 to Oct. 2007, were enrolled in this study (all patients signed informed consent), including 12 with ovarian cancer, 9 with renal cancer, 8 with lung cancer, 8 with breast cancer, 2 with colon cancer, 2 with gastric cancer, and 1 with melanoma. The donors were healthy direct relatives of the patients; the donors' haplo-PBSCs were mobilized, collected, and activated by rhIL-2 in vitro. The clinical efficacy and side effects of haplo-PBSCs therapy were assessed by CT/PET-CT scanning, RESIST standard, KPS score, and clinical response rates, etc. Results: All 42 patients received one episode of haplo-PBSCs treatment. The progression-free survivals (PFS) were 6 months and the clinical beneficial rate (CR+PR+SD) was 73.8%. The beneficial rate of life quality was 76.2% and the KPS increased by 20 (0-30) points on average after haplo-PBSCs treatment. The patients with KIR unmatched in GVH direction had better outcomes than those with KIR matched or KIR unmatched in HVG direction (P<0.05), and the clinical beneficial rate, PFS and total beneficial rate were 94.1% vs 60.0%, (13.4±1.3) vs (8.0±0.9) months, and 89.5% vs 65.2%, respectively (all P<0.05). The donor/recipient relation as the mother/child had a better outcome than that as the father/child (P<0.05). Patients with renal cancer or ovarian cancer had better outcomes than those with other cancers, with clinical beneficial rates being 90.0% and 81.8%, respectively. Conclusion: Activated haplo-PBSCs therapy can induce non-specific anti-tumor effect, and improve the clinical symptom and life quality of advanced tumor patients.

The macroscopic characteristics, tissue, caterpillar body wall and powder of Ophiocordyceps xuefengensis in different batch numbers were observed and researched by the macroscopic and microscopic identification methods. The result shows that the morphology, size, abdominal annulations of caterpillar, etc. of 0. xuefengensis are the macroscopic identification characteristics, the caterpillar body surface mycelium, body wall sculpture and crochets on abdominal legs are the microscopic identification characteristics. These characters are stable and regular discriminant features, which are proved to be the identification basis of O. xuefengensis. In addition, The characters such as crochets on abdominal legs arrange in two parallel ellipse rings, the inner crochets are long strip, and the external toes are unciform, are specific.
Animals ; Hypocreales ; cytology ; Moths ; anatomy & histology ; cytology ; microbiology

Using the HPLC-Q-TOF-MS/MS to analyze and identify the main components in the ethanol extract of Ophiocordyceps xuefengensis sp. nov. The ethanol extract of O. xuefengensis sp. nov. was preparaed by using the ultrasonic methods, the main components in the extracts were separated by using the gradient elution method with RP-HPLC, Yuexu AQ-C18 column (250 mm × 4.6 mm, 5 μm); The positive and negative electro spray ionization (ESI) source was used for determine the chromatographic effluent, the main chromatographic peaks are assigned by Q-TOF. Based on the standards of MS/MS and compared with the reference results, 28 compounds, containing mannitol, adenosine, ergosterol, sitosterol, amino acid, fatty acid, and sugar, were identified. HPLC-ESI-TOF-MS/MS method can qualitatively analyze the main components in O. xuefengensis sp. nov. from the retention time, UV spectrum, precious relative molecular weight, formula, secondary fragment ions, and so on. It is an effective and fast analysis method for determining the active compounds in O. xuefengensis sp. nov.

Objective: To establish the fingerprint of Lonicera japonica superfine powder with different polarity and study its correlation of antibacterial activity. Methods: Six different polarity extracts were obtained using the solvent system extraction and then were detected by HPLC with gradient elution. Pharmacological activities were detected by antibiotic indexes in vitro. The correlation between the pharmacological indexes and the information of HPLC fingerprint was discovered by multiple linear regression analysis and compound groups related to pharmacological indexes were obtained. Results: Antibacterial effect fingerprint of L. japonica can be established by HPLC fingerprint of ethyl acetate fraction. Conclusion: The method developed is practical and can be used for evaluating the quality control of L. japonica powder.

Objective To evaluate the safety and efficacy of 125I seed strands cavity brachytherapy for ureteral carcinoma.Methods To tally 10 patients with ureteral carcinoma underwent Carm CT and DSA guided percutaneous nephrostomy with 125I seed strands cavity brachytherapy.The technical success rate,complications,tumor local control rate,ureteral patency andsurvival time,and compared the Karnofsky scores,Girignon grade,pain score before and after treatment were evaluated.The dose related parameters were compared between pre-and post-treatment.Results 125I seed strands implantation was successfully completed in all patients with technical success rate of 100％.The mean procedure time was (12.3 ±3.8) min.No severe complications such as ureteral perforation,infection,severe bleeding occurred.Local tumor response was CR in 4 cases and PR in 6 cases,showing local control efficiency (CR + PR) 100％ after 2-3 months.Ureteral patency rate was 50％ (5/10).Postprocedure Karnofsky scores,Girignon grades,and pain scores were significantly improved (Z =-2.72,-2.88,-2.83,P＜0.01).The average follow-up time was (14.6 ±6.5) months (5-25 months),tumor progression was observed in 3 cases,stable disease in 7 cases.Nine cases were alive and one died due to multiple organ failure.The differences of D90％,mPD,V100％,V150％,V200％,CI,EI,HI between the preand post-treatment were not statistically significant (P ＞ 0.05).Conclusions 125I seed strands cavity brachytherapy for ureteral carcinoma is an effective and safe procedure without serious complications,and an effective alternative treatment for patients who are unable to undergoor refuse surgery.

Objective: To investigate the effect of RetroNectin on CIKs cells and the related mechanisms. Methods: Peripheral blood mononuclear cells (PBMC) were collected from patients and divided into two groups: group Ⅰ and group Ⅱ. Samples in group Ⅰ were seeded into culture flask precoated with RetreNec-tin and CD3mAb to induce CIKs. While samples in group Ⅱ were seeded into common culture flask. The pro-liferation of CIKs was detected by cytometric analysis. The cytotoxic activity of CIKs was determined by LDH assays. The phenotype changes and cell cycle of CIKs were identified by flow cytometry. The apoptosis of cells was detected by Annexin V/PI. Western blot was employed to detect the level of protein Vav1. The CD49d and CD49e were blocked by anti-CD49d and anti-CD49e and the proliferation of cells was tested by cytometric analysis after the blockage. The phenotype changes of cells were identified by flow cytometry after the blockage. Results: RetroNectin enhanced the proliferation of CIKs (P<0.05). Flow cytometric analysis showed that RetroNectin significantly increased the number of CD25+ T cells (P<0.05). RN-CIK was more ac-tive than CIK in killing HCT-8 cell lines in vitro (P<0.05). RetroNectin could block the CIKs at G_1 phase (P<0.05) and resist apoptosis. There was no significant difference in the proliferation between the two groups af-ter the blockage with CD49d and CD49e (P>0.05). The expression of protein Vavl was associated with CD25+T cells. Conclusion: RetroNectin enhances the proliferation of CIKs by influencing the cell cycle, resist-ing apoptosis possibly through the site of CD49d and CD49e, and inducing T cell activation as the second sig-naling through Vav1.

Methods for analyses of protein-protein interactions include: yeast two hybrid (Y2H), phage dis- play (PD), co-immunoprecipitation (Co-IP), glutathione S-transferase pull-down (GST pull-down), cellular co-localization, far-western blotting, virus overlay protein binding assay (VOPBA), surface plasmon resonance (SPR), and fluorescence resonance energy transfer (FRET). Technologies for the detection of protein-nucleic acid interactions include: yeast one hybrid (Y1H), chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), Southwestern blotting, reporter gene, Co-IP, GST pull-down, and PD. These methods are often used in the study of the human enterovirus A71 (EV-A71) by our research team. Reviews in the Chinese literature in this field are lacking, so we reviewed applications of these methods in the study of EV-A71. This review may impart important knowledge in the research of other viruses with regard to protein-protein and protein-nucleic acid interactions.
Electrophoretic Mobility Shift Assay ; Enterovirus A, Human ; chemistry ; genetics ; metabolism ; Fluorescence Resonance Energy Transfer ; RNA, Viral ; metabolism ; Two-Hybrid System Techniques ; Viral Proteins ; metabolism

OBJECTIVETo enlarge the ratio of length to width of a prefabricated random skin flap in a short period, in order to meet a special clinical need.

METHODSA white rat and a white mini-pig was chosen for an animal model for the experiment. They were divided into the experimental group and the control. The prefabricated flap was formed as the planned design. In the experimental group, the flaps were treated with a desired pressure in certain intervals at the planned part of the flap. We estimated and inspected the digitized perfusion of flap microcirculation, and made overcong and wide random skin flap survive within 24 hours, and this technique was finally used for the clinical wound treatment.

RESULTSAs expected, the survival area of the flap in the experimental group was significantly larger than the control. This technique was successfully used to repair the defects of the special sites in plastic and reconstructive surgery.

CONCLUSIONThe method of quickly prefabricated random flap could enlarge the ratio of length to width of the flap and put the flap in shape in short time. With the digitized estimating and inspecting, this method could obviously improve the survival area of the random flap and could also be very useful for the clinical treatment.

Adult ; Animals ; Dermatologic Surgical Procedures ; Female ; Humans ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Skin ; metabolism ; pathology ; Skin Transplantation ; methods ; Surgical Flaps ; supply & distribution ; Time Factors ; Transplantation, Heterologous ; Treatment Outcome

Objective:To explore the clinical application value of the dual-layer detector spectral CTA in evaluation of brain perfusion impairment in patients with acute ischemic stroke.Methods:Clinical and imaging data of 35 patients with acute ischemic stroke in Weihai Central Hospital from March 2020 to October 2020 were reviewed retrospectively. All patients underwent head and neck spectral CTA examination and dynamic cerebral perfusion CT examination with dual-layer detector spectral CT. The iodine density map and effective atomic number map were reconstructed using CTA data, and the iodine density and effective atomic number, as well as the cerebral blood volume (CBV) and cerebral blood flow (CBF) values of the hypoperfusion area and the contralateral side were measured and compared; the areas of brain hypoperfusion regions were measured. Pearson′s correlation coefficient was used to analyze the correlation between iodine density values and CBV values, iodine density values and CBF values, effective atomic number values and CBV values, effective atomic number values and CBF values, as well as hypoperfusion area shown on CTA images and displayed on CTP-CBF map.Results:Of all the 35 patients, the iodine density value [(0.22±0.07) mg/ml], effective atomic number value (7.38±0.05), CBV value [(1.9±0.7) ml/100 g] and CBF value [(15.1±5.9) ml/(100 g·min)] of the hypoperfusion area were significantly lower than those of the healthy side [iodine density value (0.44±0.10) mg/ml, effective atomic number value (7.52±0.06), CBV value (3.4±0.7) ml/100 g, CBF value (57±27) ml/(100 g·min); t values were -14.7, -14.5, -11.2, -9.7, respectively, all P<0.001]. No significant difference was found between the hypoperfusion area shown on spectral CTA [(2 292±1 393) mm 2] and shown on CTP-CBF map [(2 290±1 359) mm 2] ( t=-0.076, P=0.944). There was a positive correlation between iodine density value and CBV (affected side: r=0.350, P=0.039, healthy side: r=0.551, P=0.001); a positive correlation was also found between effective atomic number value and CBV (affected side: r=0.488, P=0.003, healthy side: r=0.552, P=0.001); and there was a strong positive correlation between the hypoperfusion area on CTA and that on CTP-CBF ( r=0.993, P<0.001). Conclusion:Dual-layer detector spectral CTA can provide the “one-stop” assessement including head and neck vascular evaluation, as well as the hypoperfution area measument, which can be an alternative rapid method for evaluation of patients with acute ischemic stroke.