Objective: To discriminate traditional She medicine Gegongniugen (the root of Rubus chingii Hu) from its confusable species of genus Rubus based on ITS2 barcode, and to guarantee the medicine quality and clinical effect of Gegongniugen. Methods: A total of 140 samples of R. corchorifolius L. f., R. hirsutus Thunb., R. parvifolius L., R. coreanus Miq., R. buergeri Miq., R.tsangorum Hand. -Mazz., R. trianthus Focke, R. jiangxiensis Z. X. Yu, W. T. Ji et H. Zheng were collected from Zhejiang, Guangxi, Jiangxi, and Anhui provinces. Genomic DNA was extracted from the collected samples, followed by ITS2 sequences amplification through PCR, bi-directional sequencing, assembly and annotation based on HMMer method. At the same time, 37 other sequences of corresponding species were downloaded from GenBank. Sequence characteristics such as lengths, GC contents, and variable sites were analyzed by Gene Tool software while genetic distance calculation and build-up of neighbor-joining (NJ) phylogenetic tree were carried out through Clustal X and MEGA 7.0. Results: The lengths of ITS2 sequences of Gegongniugen and its nine confusable species of genus Rubus ranged from 211 to 213 bp with GC content of 52.1%-58.0%; These ITS2 sequences had 14 mutation sites and one insertion/deletion site; The average intra-specific K2P genetic distance of Gegongniugen was 0.007 2, which was significantly lower than the respective inter-specific K2P genetic distance between Gegongniugen and its confusable species of genus Rubus. The genetic distantes between Gegongniugen and R. corchorifolius or Gegongniugen and R. jiangxiensis were minimum, while that between Gegongniugen and R. hirsutus was maxmum. It was obvious in the N-J phylogenetic tree that the ITS2 sequences of Gegongniugen formed into one separate clade, which could be successfully distinguished other nine species. Conclusion: ITS2 sequence can be used as an effective DNA marker to discriminate the original plants of Gegongniugen and its confusable species of genus Rubus, which provided an important molecular evidence for the identification of traditional She medicines and contributed to the information supervision of traditional She medicine market.

In this study, pEGFP-N1 was chosen as the reporter plasmid and transferred into Ctenopharyngodon idellus kidney (CIK) cells by electroporation, and the optimal electroporation conditions were determined by testing the transfection efficiency with different voltages, pulse times, plasmid amounts, and numbers of shocks. The results showed that the maximum electroporation efficiency was achieved under the following conditions in a 0.2 cm electroporation cuvette containing CIK cells (1.5 x 10(7)/mL, 200 microl): electric voltage 200 V, pulse time 45 ms, plasmid 30 microg, and one electric shock. The total genomic RNA of grass carp reovirus (GCRV) was extracted in this experiment and reversely transcribed into cDNA, which was used to amplify the gene segment of GCRV non-structural protein NS26 using designed specific primers. The PCR product was recombined into pEGFP-N1 vector. The fusion protein EGFP-NS26 was successfully and efficiently expressed in the CIK cells by electroporation, which was confirmed by both fluorescent imaging and Western blot analysis. This experiment laid a foundation for further functional studies of the non-structural protein NS26 of GCRV.
Animals ; Cell Line ; Cyprinidae ; Electroporation ; Fish Diseases ; virology ; Gene Expression ; Kidney ; virology ; Reoviridae ; genetics ; physiology ; Reoviridae Infections ; veterinary ; virology ; Viral Nonstructural Proteins ; genetics ; metabolism

OBJECTIVE:To compare home-made mu'an-eye-gel(acyclovir plus honey) with commercial aciclovir(ACV)-eye-gel in releasing drug characteristics in vitro.METHODS:The in vitro drug release test was conducted by the third method of dissolution determination stated in Chinese Pharmacopeia together with bag filler method.The cumulative drug-releasing percentage and the acyclovir amount in mu'an-eye-gel versus ACV-eye-gel were determined by UV spec-trophotometry,and the accumulative releasing drug percentages of the two preparations were computed and their drug release behaviors w ere compared.RESULTS:The in vitro releasing behaviors of mu'an-eye-gel followed the Weibull kinetic equa-tion,however the vitro releasing behavior of commercial ACV-eye-gel followed the zero order kinetic equation,and the T80%and Q8 h had statistical significances between(mu'an-eye-gel:T80%=3.156?0.013(h),Q8 h=93.28?0.010(%);ACV-eye-gel:T80%=10.16?0.009(h),Q8 h=67.85?0.025(%)) 2 kinds of preparation.CONCLUSION:Mu'an-eye-gel is superior to the commercial ACV ophthalmic gel in both releasing velocity and accumulative drug release percentage.

The paper is about the systematic studies of biological characteristics of 15 stains rhizobia isolated purified from Acacia confusa grew in Guangxi karst environment.The results showed that there were typical characteristics of rhizobia.there were negative reaction about use of 3-ketolactose and beef extract peptone nutrient agar medium,and positive reaction about use of starch and citrate medium,and produce acid in reaction of BTB and litmus milk medium,(NH4)2HPO4 was used as nitrogen sources and both four monosaccharides and three disaccharides could be utilized as carbon sources in 15 strains rhizobia isolated Acacia confuse.Among the 15 strains for the tests,11 strains could deoxidize the nitrate of medium into nitrite,14 strains could grow well on NaCl solution concentration 3.0 %～4.0 %,14 strains could grow at 39℃,13 strains may grow on highest pH12 and 4 strains on lowest pH4 cultrue medium.15 strains can grow in 10% and 11 strains in 10%～30% of CaCO3 solution concentration.

Objective To study the feasibility of using heat and moisture exchangers (HME)as an alternative to heated humidifiers (HH) in patients undergoing mechanical ventilation. Methods 266 pa-tients with mechanical ventilation admitted to our ICU over the recent 3 years were allocated to the experi-mental group (humidification with a heat and moisture exchanger) and the control group (with heated hu-midifier), and the effect of humidification, the reserved time of artificial airway, the time on mechanical yen-tilation, the time of stay in ICU, the ineidenee of ventilator-associated pneumonia (VAP) and the mor-tality rate were comparatively studied and analyzed. Results Significant differences were found between the experimental and the control group in effect of humidification, insufficiency of humidification or excessive hu-midification, airway spasm and time on mechanical ventilation and time of stay in ICU. The incidence of VAP in the control group was significantly higher than that in the experimental group. There were no significant dif-ference between the two groups in the reserved time of artificial airway and the mortality rate. There were no accident of humidification occurred in the experimental group while there were one case complicated with air-way burn and 11 eases complicated with choking with water in the control group. Conclusions We conclude that HH can be replaced by HME on mechanical ventilation while disease evolution and effect of humidification should be monitored closely and keep HME unobstructed.

OBJECTIVE: Explore the model of universal NICU newborns' hearing screening in high-risk neonates, preliminary understanding factor of hearing damage. METHOD: Transient evoked otoacoustic emissions (TEOAE) and automatic auditory brainstem response (AABR) were used to detect newborns' hearing in 13 315 objects, that is newborns' hearing screening in NICU with TEOAE test who not pass, 42 days after will use AABR rescreening. Children's Hearing Center of Guangxi Child Health Hospital will diagnose the newborns that did not pass in 3 months. RESULT: In these 13 315 newborns, 5 151 subjects who did not pass the initial screening, 1910 subjects who also did not pass after 42 days, 1167 subjects cannot pass the rescreening after 3 months, 642 subjects were diagnosed congenital hearing impairment by Brainstem Auditory Evoked Potential Test, the rate is 4.82%. CONCLUSION TEOAE and AABR are the suitable model of universal newborns' hearing screening in high-risk neonates.
Evoked Potentials, Auditory, Brain Stem ; Female ; Follow-Up Studies ; Hearing Tests ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; Male ; Neonatal Screening

OBJECTIVETo evaluate the efficacy and safety of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) hydrogel in wound healing in patients with deep partial thickness burn.

METHODSThe study was a multicenter, randomized, double-blind, placebo-controlled parallel clinical trial. Three hundred and twenty-one patients (302 cases finally fulfilled the protocol) with deep partial thickness burn were divided into A group (n = 200, with treatment of rhGM-CSF hydrogel, 100 microg/10 g/100 cm2/d), C group (n = 102,with treatment of placebo). Side-effect, systemic condition, wound healing time, wound healing rate, and total effective rate at different time points were observed.

RESULTSThere were no obvious differences in vital signs, wound secretion, wound edge reaction, blood and urine routine, liver and kidney function between two groups (P > 0.05). No side-effect was observed. The median wound healing time was 17 days in A group, which was obviously shorter than that in C group (20 days, P < 0.01). The mean wound healing rate in A group was 24.5%, 70.5%, 95.3%, 99.6% respectively on 8th, 14th, 20th, 28th day after treatment, which were obviously higher than that in C group (15.1%, 51.4%, 84.6%, 97.1%, respectively, P < 0.01). The total effective rates in A group on 8th, 14th, 20th day after treatment were also higher than that in C group (P < 0.01).

CONCLUSIONrhGM-CSF hydrogel can significantly accelerate wound healing in patients with deep partial thickness burn with certain safety.

Burns ; drug therapy ; Double-Blind Method ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; adverse effects ; therapeutic use ; Humans ; Hydrogels ; therapeutic use ; Male ; Placebos ; Recombinant Proteins ; Wound Healing

OBJECTIVETo analyze clinical features of 4 families with hereditary hemorrhagic telangiectasia (HHT) and potential mutations of ENG, ACVRL1 and SMAD4 genes.

METHODSFour unrelated HHT patients and their affected family members were analyzed. All exons and flanking regions of ENG, ACVRL1 and SMAD4 genes were analyzed with PCR and direct sequencing and multiplex ligation-dependent probe amplification (MLPA) methods.

RESULTSEleven patients from the 4 families were enrolled in this study. Two ENG and 1 ACVRL1 mutations were identified, among which an ENG mutation (c.207G>A; p.L69L) and an ACVRL1 mutation (c.817C>T; p.L273L) have been previously reported. In addition, a novel ENG mutation (c.1004A>T; p.Q335L) has been found in 3 different families. Similar mutations were not detected in 200 healthy individuals. No mutations of ENG, ACVRL1 and SMAD4 were found in the fourth family.

CONCLUSIONA novel mutation c.1004A>T (p. Q335L) of ENG has been identified in patients with HHT. And there is significant phenotypic variability and genetic heterogeneity with the disease.

Activin Receptors, Type II ; genetics ; Adolescent ; Adult ; Amino Acid Sequence ; Antigens, CD ; genetics ; Endoglin ; Female ; Genetic Testing ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Receptors, Cell Surface ; genetics ; Smad4 Protein ; genetics ; Telangiectasia, Hereditary Hemorrhagic ; diagnosis ; genetics

Objective To investigate the efficacy of an anti-vascular endothelial growth factor (VEGF) antibody,MIL60,in inhibiting corneal neovascularization (CoNV) formation in a rat model of alkali cauterization and its involved mechanisms.Methods Rat CoNV model induced by alkali burn was founded in the right eyes,and then 72 cases were randomly divided into four groups according to the subconjunctival administration of medicine next day after the successful establishment of this model:25mg· mL-1 MIL60 group,dexamethasone group,MIL60 solvent group and NaCl group.Then CoNV was observed for recording the its length and the involved area using digital photograph.Next the rats were sacrificed on day 7,14,21 and 28,followed by the collection of rats' cornea for HE and immunohistochemical staining to analyze the protein expression of VEGF,VEGF receptor-1 (VEGFR-1),VEGFR-2 and matrix metallopeptidase-9 (MMP-9).Results At each time point,the area and length of CoNV in the 25 mg· mL-1 MIL60 and dexamethasone group were significantly less than those in the MIL60 solvent and NaC1 group,and the differences were statistically significant (all P ＜0.01),and 25 mg· mL-1 MIL60 group had the similar CoNV area and length with the dexamethasone group (all P ＞ 0.05).Moreover,HE and immunohistochemical staining showed that MIL60 could inhibit the protein expression of VEGF,VEGFR-1,VEGFR-2 and MMP-9,which could explain its effective anti-angiogenic activity.Conclusion Subconjunctival administration of MIL60 can significantly inhibit corneal neovascularization formation and alleviate the inflammation in rats suffered from alkali burn.

Objective:To investigate the expression of Fas、FasL and the apoptosis of liver cancer cell line HepG2 transfected with LIGHT and IFN-? gene mediated by Cationic liposome.Methods:HepG2 cells were divided into two groups(the solo transfection of LIGHT gene and the combined transfection of LIGHT and IFN-? genes) and the control groups(no transfection).HepG2 cells were cellected at 12h,24h and 48h after transfection.The apoptosis of HepG2 cells and the expression of Fas and FasL of the HepG2 cells were investigated with flow cytometry.Results:After transfection,the apoptosis of HepG2 cells increased,and the apoptosis of combined transfection group was higher than the solo transfection of LIGHT(P