Collagen-coated agarose-chitosan scaffold used as a skin substitute.

A single biomaterial is disadvantageous for constructing skin in vitro, so a mixed biomaterial is more conducive to skin research. In this study, agarose-chitosan scaffolds with a final concentration of 4% were constructed by freeze-drying, in which the concentration ratios of agarose to chitosan were 1:3, 2:2, and 3:1. The scaffolds were coated with a 3 mg/ml collagen solution, and the mechanical properties were evaluated by studying density, porosity, swelling rate, and degradation rate. The results demonstrated that the agarose-chitosan scaffolds were porous, with porosity reaching 93%. Their densities ranged from 0.1 to 0.16 g/cm3 . Analysis of Young's modulus showed that the mechanical properties of the agarose-chitosan scaffolds were significantly enhanced when the agarose content in the agarose-chitosan scaffolds was increased. Moreover, the density and Young's modulus of the agarose-chitosan scaffolds of different concentration ratios were significantly different (p < 0.01). These scaffolds can withstand a certain amount of external pressure, such as that of human skin, making them more suitable for further skin replacement research. In addition, the results of thiazolyl blue tetrazolium bromide (MTT) cell assay and immunofluorescence staining showed that the collagen-coated agarose-chitosan scaffolds were conducive to keratinocyte proliferation and differentiation. The MTT results revealed significant differences between the agarose-chitosan scaffolds coated with collagen and the agarose-chitosan scaffolds without collagen (p < 0.05). This study provides the potential for in vitro skin research and applications.

3D Ionic Olefin-Linked Conjugated Microporous Polymers for Selective Detection and Removal of TcO4-/ReO4- from Wastewater.

Technetium (99Tc) is a highly toxic radioactive nuclear wastewater contaminant. Real-time detection of 99Tc is very difficult due to its difficult-to-complex nature. Herein, a novel three-dimensional ionic olefin-linked conjugated microporous polymer (TFPM-EP-Br) is constructed using tetrakis(4-aldehyde phenyl)methane (TFPM) as the central monomer. The unique cationic cavity and highly hydrophobic framework enable TFPM-EP-Br to act as a fluorescent sensor for TcO4-. The fluorophores of TFPM-EP-Br can be quenched due to electron transfer from TFPM-EP-Br to TcO4- and the formation of strongly nonfluorescent complexes. Meanwhile, the regular pore channels are beneficial for the fast mass transfer of TcO4-, resulting in an ultrafast response time (less than 2 s) with an ultralow detection limit (33.3 nM). In addition, the ultrahigh specific surface area enables TFPM-EP-Br to combine the ability to synergistically detect and remove radioactive 99Tc. From this perspective, the novel conjugated microporous polymer has made a breakthrough in the detection and extraction of radioactive contaminants.

Physicochemical properties, antioxidant activities and in vitro sustained release behaviour of co-encapsulated liposomes as vehicle for vitamin E and ß-carotene.

BACKGROUND: In this study the potential of liposomes as a vitamin E (VE) and ß-carotene (ßC) delivery system was examined. The co-encapsulated liposomes of ßC and VE (L-VE-ßC) were prepared and characterized. Their antioxidant properties were evaluated by free radical scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), hydroxyl radical and lipid peroxidation assay. The in vitro sustained release behaviour was then investigated and discussed. RESULTS: VE and ßC were co-encapsulated in liposomes with high encapsulation efficiency, up to 92.49% and 86.16% for ßC and VE, respectively. The antioxidant activities of L-VE-ßC samples were significantly higher than that of single loaded liposome. Among different ratios of VE/ßC, L-VE-ßC at 5:3 exhibited the highest radical scavenging rates, with 66.80%, 56.58% and 34.39% for DPPH, ABTS and OH radical, respectively. L-VE-ßC samples also had a good ability to inhibit lipid peroxidation, especially the sample with ratios of VE/ßC at 5:3 and 3:1. In simulated gastrointestinal release, L-VE-ßC exhibited an excellent sustained release behaviour in SGF with the accumulated rate at about 20%, while the release rate in SIF increased to over 80%, where they should be absorbed. The release kinetics analysis indicated that ßC was released in the Higuchi model in stomach, and the Korsmeyr-Peppas model in intestine. CONCLUSION: Compared to single loaded liposomes, the combined-loaded liposomes exhibited higher antioxidant activity and bioavailability, suggesting the potential applications in functional foods. © 2022 Society of Chemical Industry.

A novel bicistronic expression system composed of the intraflagellar transport protein gene ift25 and FMDV 2A sequence directs robust nuclear gene expression in Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii offers a great promise for large-scale production of multiple recombinant proteins of pharmaceutical and industrial interest. However, the nuclear-encoding transgenes usually are expressed at a low level, which severely hampers the use of this alga in molecular farming. In this study, the promoter of the endogenous intraflagellar transport 25 (IFT25) gene of C. reinhardtii was tested for its ability to drive the expression of green fluorescent protein (GFP), which functions as a readout for target gene expression. IFT25 promoter (IFT25P) alone was not able to drive GFP expression to a detectable level. IFT25P, however, can drive robust IFT25-GFP fusion protein expression when the intron-containing IFT25 gene was inserted between IFT25P and GFP cDNA. When an extended version of foot-and-mouth virus 2A protease (2AE) sequence was further inserted between the intron-containing IFT25 gene and the GFP cDNA, discrete GFP protein was observed to release from the IFT25-2AE-GFP polyprotein via 2A self-cleaving with a cleavage efficacy of approximately 99%. The monomer GFP was accumulated to a level of as high as 0.68% of total soluble proteins. To test whether the newly developed bicistronic IFT25P-IFT25-2AE expression system can be used to overexpress heterologous proteins of different origins and sizes, we inserted codon-optimized cDNAs encoding a Trichoderma reesei xylanase1 (25 kDa) and a Lachnospiraceae bacterium ND2006 type V CRISPR-Cas protein LbCpf1 (147 kDa) to the vector and found that the production of xylanase1 and LbCpf1 was as high as 0.69 and 0.49% of total soluble protein. Our result showed that IFT25P-IFT25-2AE system is more efficient to drive nuclear gene expression in C. reinhardtii than other conventionally used promoters, thus representing a novel efficient recombinant protein expression tool and has the potential to be scaled for commercial production of nuclear-encoded recombinant proteins of different sizes and origins in C. reinhardtii.

Genotoxic effects of old landfill leachate on HepG2 cells after nitration/ultrafiltration/reverse osmosis membrane treatment process.

Toxicity assessment of nitration/ultrafiltration/reverse osmosis (nitration/UF/RO) project, which has recently been widely used as an efficient process with applications in practical leachate treatment, was very limited. In the present study, DNA damage of leachates was investigated before and after the nitration/UF/RO process by a battery of assays with human hepatoma cells. Methyletrazolium assay showed a high cytotoxicity of 97.1% after being exposed to the highest concentration of raw leachate for 24 h, and a cytotoxicity of 26% in effluent at a concentration of 30% (v/v). Both comet assay (24 h) and γH2AX flow cytometer assay (3 h) showed increased levels of DNA damage in cells exposed to raw leachate and after nitration/UF-treated leachate followed by a significant increase of 7-ethoxyresorufin-O-deethylase activity. However, the effluent after nitration/UF/RO treatment showed no significant difference compared to negative control for γH2AX flow cytometer assay but slight DNA damage at concentrations of 20% and 30% (v/v) as well as increase of 7-ethoxyresorufin-O-deethylase. Analysis showed that nitration/UF/RO process exhibited high removal of physicochemical indexes and significant reduction of toxic and genotoxic effects of leachate, but still demands an improvement to reduce all possible negative risks to the environment and humans. Copyright © 2017 John Wiley & Sons, Ltd.

[Efficacy and Safety Evaluation of Liujin Runzao Concentrated Decoction in Treating Primary Sjögren's Syndrome].

Objective To evaluate the efficacy and safety of Liujin Runzao Concentrated Decoction (LRCD) for the treatment of primary Sjögren's syndrome (pSS). Methods Forty pSS patients with fluid depletion and distribution obstacles syndrome (FDDOS) were randomly assigned to the experimen- tal group and the control group according to 1:1 proportion. All patients received standard therapy: Radix Paeoniae alba total glycosides 600 mg, twice per day. Patients in the experimental group additionally took LRCD, 30 mL each time, twice per day. The therapeutic course for all was 4 weeks, and two courses for all. The improvement of dry mouth and dry eyes were comprehensively evaluated. Each outcome of composite index constitutions (integrals of dry eyes and dry mouth, salivary flow rate, Schirmer test) was respectively reported. Schirmer test and salivary flow rate were determined as well. Score of TCM syndrome, blood sedimentation,'immunoglobulin, and adverse drug reactions were observed. Results The effective rate of comprehensive effect for dry eyes and dry mouth improvement at the end of 8 weeks was 80% in the experimental group and 35% in the control group, with statistical difference (X² =8. 286, P <0. 05). As for the composition of comprehensive effect for dry eyes and dry mouth improvement: The score for dry eyes and dry mouth decreased in the two groups more after treatment than before treatment. The difference in pre-post treatment score for dry eyes and dry mouth at week 8 was higher in the experimental group than in the control group. The difference in pre-post treatment score at week 8 was 1. 71 (95% Cl: -0. 37 -3. 78) between the two groups (P <0. 05). The difference in pre-post treatment Schirmer test and salivary flow rate at week 8 was higher in the experimental group than in the control group, but with on statistical difference (P >0. 05). The difference in pre-post treatment Schirmer test and salivary flow rate at week 8 was 2. 74 mL/15 min (95% Cl: 0. 49 -4.98) and 0. 13 mm/5 min (95% Cl: 0. 92 -1. 23) between the two groups (P <0. 05). The score of TCM syndrome decreased more in the two groups, as compared with before treatment. The difference in pre-post treatment score of TCM syn- drome at week 8 was 1. 71 (95% CI: -1. 40 -4. 81) between the two groups (P >0. 05). One case of uri- nary tract infections occurred in the control group, while no obvious adverse event occurred in the exper- imental group. Conclusion Standard treatment combined LRCD showed better comprehensive effect for dry eyes and dry mouth in pSS patients with FDDOS, and was more safe.

Preparation and characterization of water vapor-responsive methylcellulose-polyethylene glycol-400 composite membranes and an indication of freshness of shiitake mushrooms.

Intelligent packaging with freshness indication capability can help consumers purchase fresh food. However, current research primarily focuses on carbon dioxide-sensitive intelligent packaging, with limited research on water vapor-sensitive indication packaging. In this study, the water vapor-sensitive indicator membrane was prepared and used to determine the freshness of mushrooms. The results of this study showed that the water permeability of the indicator membrane decreased from 33.17 % to 21.59 % with the increase of Polyethylene glycol-400(PEG-400) content in methylcellulose(MC) membrane, and the contact angle of the indicator membrane increased from 87 % to 98 % with the addition of PEG-400. The addition of plasticizer PEG-400 increased the hydrophobicity of the indicator film, which could be attributed to the improvement of the molecular arrangement and crystallinity of the indicator film by the addition of PEG-400. After encountering water, the transparency of the indicator membrane changes from completely opaque (white) to transparent. Addition of PEG-400 reduces the rate of change in the transparency of the indicator membrane. The indicator membrane was successfully used to indicate the freshness of mushrooms and effectively reflected the freshness of mushrooms during storage. This technology could be applied to measure the freshness of other foods.

Design strategy of self-assembled BC@MIL-100(Fe) composite membrane for the efficient removal of diclofenac sodium from water.

Pharmaceuticals and personal care products (PPCPs) as typical emerging pollutant have attracted extensive attention due to the risks to human health and environment. As a kind of harmful PPCPs, diclofenac sodium (DCF) has been frequently detected in water environment, which needs to be removed effectively. Herein, we successfully fabricated network structure composite membranes consisting of one-dimensional (1D) well-defined core-shell bacterial cellulose (BC)@MIL-100 (Fe) nanofibers by a simple but effective step-by-step strategy. The BC@MIL-100(Fe) composite membrane has three-dimensional network utilizing bacterial cellulose nanofiber as template, which was observed as the MIL-100(Fe) grew on the nanofiber uniformly and obvious core-shell structure. Impressively, the BC@MIL-100(Fe) not only possessed favorable architecture but also behaved good properties to adsorb DCF from water. As we expected, the as-obtained BC@MIL-100(Fe) composite membrane exhibited convenient recycling, high chemical stability, and short equilibrium time (30 min) with high adsorption capacity (296 mg g-1). Strikingly, in low DCF concentration solution, BC@MIL-100(Fe) composite showed high adsorption efficiency in periodic test, which can still reach 70% after five successive adsorptions without desorption. The results demonstrated that the adsorption mechanism may involve π-π interaction, H-bond interaction, and electrostatic interaction. This work proposes the new finding to understand the removing of DCF from water environment.

Saliva proteome of partially- and fully-engorged adult female Haemaphysalis flava ticks.

Tick saliva is a reservoir of bioactive proteins. Saliva protein compositions change dynamically during blood-feeding. Decipherment of protein profiles in different blood-feeding stages may bring deeper insight into tick feeding physiology and provide targets for immunologic control alternatives. However, having the infancy of tick genome sequencing, assembly, annotation, and limited knowledge of tick salivary proteins restrain the data interpretation. Here, we aimed to depict the saliva protein profile in partially- (PE) and fully-engorged (FE) Haemaphysalis flava ticks, with a special focus on the analysis of those uncharacterized proteins. Saliva was collected from PE and FE adult female H. flava ticks. Saliva proteins were analyzed by high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS-MS). MS data were searched against an in-house salivary gland transcriptome library for identification of tick-derived proteins. Abundances of proteins were compared between PE and FE ticks. The uncharacterized proteins detected in saliva were further bioinformatically analyzed. In total, 614 proteins were identified including 94 host proteins and 520 tick-derived proteins. The 226 tick-derived high-confidence proteins were classified into 10 categories: transporters, enzymes, protease inhibitors, immunity-related proteins, lipocalins, glycine-rich proteins, muscle proteins, secreted proteins, uncharacterized proteins and others. A total of 98 proteins were shared in both PE and FE with 74 only in PE and 54 only in FE. Abundances of 24 shared proteins were significantly higher in PE. The profile of top 15 most abundant proteins was also different between PE and FE ticks. The 65 uncharacterized proteins detected in tick saliva were branched into subclusters 1 A, 1B, 2, 3 A, 3B and 3 C based on particular motifs like RGD, LRR, indicating their diverse predicted functions like anti-coagulation, regulation of innate immune, or other functions. This study provides and compares saliva proteomes of H. flava ticks in two feeding stages with special cluster analysis on the uncharacterized proteins. Further investigations are needed to confirm the roles of these uncharacterized proteins in ticks.

Clinical Efficacy and Safety of Foldable Capsular Vitreous Body Implant Surgery in 22 Cases of Silicone Oil-dependent Eyes.

OBJECTIVE: This study aimed to evaluate the clinical efficacy and safety of foldable capsular vitreous body (FCVB) implant surgery in silicone oil-dependent eyes. METHODS: A total of 22 participants with silicone oil-dependent eyes who received treatment with FCVB implant surgery between January 2019 and June 2020 were included in this retrospective study. The intraocular pressure (IOP), best-corrected visual acuity (BCVA), demographic data, and any recorded complications were evaluated. RESULTS: The postoperative IOP (12.73±4.20 mmHg) was significantly improved (P=0.03) compared to the preoperative IOP (10.23±3.69 mmHg) (the main endpoint). There was no significant difference (P=0.33) in the final BCVA preoperation and 3rd month postoperation (the secondary endpoint). The most common postoperative complication was hyphema. Other common postoperative complications included corneal opacity, a shallow anterior chamber, and a low IOP. CONCLUSION: FCVB implant surgery is a safe and effective method for treating silicone oil-dependent eyes; however, attention should be paid to the prevention and timely treatment of complications.

Biocompatible chitosan/polyethylene glycol/multi-walled carbon nanotube composite scaffolds for neural tissue engineering.

Carbon nanotube (CNT) composite materials are very attractive for use in neural tissue engineering and biosensor coatings. CNT scaffolds are excellent mimics of extracellular matrix due to their hydrophilicity, viscosity, and biocompatibility. CNTs can also impart conductivity to other insulating materials, improve mechanical stability, guide neuronal cell behavior, and trigger axon regeneration. The performance of chitosan (CS)/polyethylene glycol (PEG) composite scaffolds could be optimized by introducing multi-walled CNTs (MWCNTs). CS/PEG/CNT composite scaffolds with CNT content of 1%, 3%, and 5% (1%=0.01 g/mL) were prepared by freeze-drying. Their physical and chemical properties and biocompatibility were evaluated. Scanning electron microscopy (SEM) showed that the composite scaffolds had a highly connected porous structure. Transmission electron microscope (TEM) and Raman spectroscopy proved that the CNTs were well dispersed in the CS/PEG matrix and combined with the CS/PEG nanofiber bundles. MWCNTs enhanced the elastic modulus of the scaffold. The porosity of the scaffolds ranged from 83% to 96%. They reached a stable water swelling state within 24 h, and swelling decreased with increasing MWCNT concentration. The electrical conductivity and cell adhesion rate of the scaffolds increased with increasing MWCNT content. Immunofluorescence showed that rat pheochromocytoma (PC12) cells grown in the scaffolds had characteristics similar to nerve cells. We measured changes in the expression of nerve cell markers by quantitative real-time polymerase chain reaction (qRT-PCR), and found that PC12 cells cultured in the scaffolds expressed growth-associated protein 43 (GAP43), nerve growth factor receptor (NGFR), and class III ß|-tubulin (TUBB3) proteins. Preliminary research showed that the prepared CS/PEG/CNT scaffold has good biocompatibility and can be further applied to neural tissue engineering research.

Construction of tissue-engineered skin with rete ridges using co-network hydrogels of gelatin methacrylated and poly(ethylene glycol) diacrylate.

Tissue-engineered skin, as a promising skin substitute, can be used for in vitro skin research and skin repair. However, most of research on tissue-engineered skin tend to ignore the rete ridges (RRs) microstructure, which enhances the adhesion between dermis and epidermis and provides a growth environment for epidermal stem cells. Here, we prepared and characterized photocurable gelatin methacrylated (GelMA) and poly(ethylene glycol) diacrylate (PEGDA) co-network hydrogels with different concentrations. Using a UV curing 3D printer, resin molds were designed and fabricated to create three-dimensional micropatterns and replicated onto GelMA-PEGDA scaffolds. Human keratinocytes (HaCaTs) and human skin fibroblasts (HSFs) were co-cultured on the hydrogel scaffold to prepare tissue-engineered skin. The results showed that 10%GelMA-2%PEGDA hydrogel provides the sufficient mechanical properties and biocompatibility to prepare a human skin model with RRs microstructure, that is, it presents excellent structural support, suitable degradation rate, good bioactivity and is suitable for long-term culturing. Digital microscope image analyses showed the micropattern was well-transferred onto the scaffold surface. Both in vitro and in vivo experiments confirmed the formation of the epidermal layer with undulating microstructure. In wound healing experiments, hydrogel can significantly accelerate wound healing. This study provides a simple and powerful way to mimic the structures of human skin and can make a contribution to skin tissue engineering and wound healing.

Adsorption of Sr(II) from water by mercerized bacterial cellulose membrane modified with EDTA.

The treatment of low-level radioactive wastewater is a critical and considerable challenge. Bacterial cellulose membrane (BCM) modified with ethylenediaminetetraacetic acid (EDTA) using (3-aminopropyl) triethoxysilane (APTES) as a crosslinker were used to remove Sr2+ in this work. SEM, XPS, and FTIR were used to characterize the morphology, structure, chemical shift, and functional groups of the as-prepared adsorbent. The synthesized BCM@APTES-EDTA presented a three-layer structure of membrane-net-membrane with nano-sized fibers (<100 nm). The adsorption of Sr2+ onto BCM@APTES-EDTA was investigated as a function of contact time and initial concentration of Sr2+. Results showed that the adsorption of Sr2+ followed the pseudo second-order kinetic model (R2 = 0.999), and fitted well with the Langmuir isotherm model (R2 = 0.996). The maximum adsorption capacity was calculated to be 44.86 mg g-1, which was comparable to other adsorbents. Additionally, the mechanism of Sr2+ adsorbed by the as-prepared adsorbent was studied through FTIR and XPS analysis, which indicated that the tertiary amines and carboxylate from grafted EDTA participated in the adsorption of Sr2+.

Anti-oral common pathogenic bacterial active acetylenic acids from Thesium chinense Turcz.

Discovery of agents for oral infectious diseases is always encouraged in natural products chemistry. A bioassay-guided isolation led to the isolation of two new acetylenic acids (1, 2) along with seven known ones (3-9) from the ethanol extract of Thesium chinense Turcz, a commonly used oral anti-bacterial and anti-inflammatory herb. Their structures were elucidated on the basis of spectroscopic and chemical evidence. Exocarpic acid (3) demonstrated the most promising activity against three tested oral pathogenic bacterial strains, Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus mutans, with minimum inhibitory concentration values of 0.86, 3.43, and 13.70 µg/mL, respectively. Compounds 1, 2, 4, 5 and 7 also showed potential activities against periodontal bacteria (P. gingivalis, F. nucleatum).

Effect of Double-Balloon Enteroscopy on Diagnosis and Treatment of Small-Bowel Diseases.

BACKGROUND: The diagnosis and treatment of small-bowel diseases is clinically difficult. The purpose of this study was to evaluate the diagnostic and therapeutic value of double-balloon enteroscopy in small-bowel diseases. METHODS: The history and outcomes of 2806 patients who underwent double-balloon enteroscopy from July 2004 to April 2017 were reviewed, which included 562 patients with obscure digestive tract bleeding, 457 patients with obscure diarrhea, 930 patients with obscure abdominal pain, 795 patients with obscure weight loss, and 62 patients with obscure intestinal obstruction. Examinations were performed through the mouth and/or anus according to the clinical symptoms and abdominal images. If a lesion was not detected through one direction, examination through the other direction was performed as necessary. Eighty-four patients with small-bowel polyps, 26 with intestinal obstruction caused by enterolith, and 18 with bleeding from Dieulafoy's lesions in the small intestine were treated endoscopically. RESULTS: A total of 2806 patients underwent double-balloon enteroscopy, and no serious complications occurred. An endoscopic approach through both the mouth and anus was used in 212 patients. Lesions were detected in 1696 patients, with a detection rate of 60.4%; the rates for obscure digestive tract bleeding, diarrhea, abdominal pain, weight loss, and intestinal obstruction were 85.9% (483/562), 73.5% (336/457), 48.2% (448/930), 49.1% (390/795), and 62.9% (39/62), respectively. For patients with small-bowel polyps who underwent endoscopic therapy, no complications such as digestive tract bleeding and perforation occurred. Intestinal obstruction with enteroliths was relieved with endoscopic lithotripsy. Among the 18 patients with bleeding from small-bowel Dieulafoy's lesions, 14 patients were controlled with endoscopic hemostasis. CONCLUSION: Double-balloon enteroscopy is useful for diagnosing and treating some small-bowel disease.

Application of double-balloon enteroscopy in the diagnosis and therapy of small intestinal diseases.

BACKGROUND: X-ray of the digestive tract and radionuclide examination could not accurately detect diseases of the small intestine. Double-balloon enteroscopy has been used to increase the detection rate of these diseases in addition to endoscopic biopsy and therapy. The purpose of this study was to determine the value of double-balloon enteroscopy in the diagnosis and treatment of diseases of the small intestine. METHODS: A total of 258 double-balloon enteroscopies via the mouth and/or anus were performed in 208 patients. If no lesion was detected on one side (mouth or anus), examination on the other side (anus or mouth) was made. If active small intestinal bleeding was detected, endoscopic hemostasis was done to treat the bleeding. RESULTS: In the 208 patients, 50 were subjected to double-balloon enteroscopy via both mouth and anus. Lesions were detected in 151 patients, giving a rate of 72.6% (151/208). The detection rates for obscure digestive tract bleeding, diarrhea, abdominal pain and weight loss were 90.2% (92/102), 64.9% (24/37), 48.5% (16/33) and 43.3% (13/30), respectively. Lesions of the 151 patients were confirmed by endoscopic biopsy, surgery, clinical studies, and follow-up. In the 102 patients with bleeding of the digestive tract, active bleeding was detected in 27 patients. Endoscopic hemostasis was successful in 25 of them (92.6%, 25/27). No serious complications occurred in all the patients, the average time for the procedure was 100 minutes. CONCLUSIONS: Double-balloon enteroscopy is safe, effective in the diagnosis of diseases of the small intestine in addition to endoscopic therapy.

Anti-viral and cytotoxic norbisabolane sesquiterpenoid glycosides from Phyllanthus emblica and their absolute configurations.

In an effort to identify anti-viral and cytotoxic compounds from Phyllanthus spp., 14 highly oxygenated norbisabolane sesquiterpenoids, phyllaemblicins H1-H14, were isolated from the roots of Phyllanthus emblica Linn, along with phyllaemblicins B and C and glochicoccinoside D. Their structures were determined on the basis of detailed spectroscopic analysis and chemical methods. Determination of absolute configurations of these compounds was facilitated by theoretical calculations of electronic circular dichroism (ECD) spectra using time-dependent density functional theory (TDDFT) for the aglycone components, and pre-column derivative/chiral HPLC analysis for the monosaccharides. The known glochicoccinoside D displayed potent activity against influenza A virus strain H3N2 and hand, foot and mouth virus EV71, with IC50 values of 4.5±0.6 and 2.6±0.7 µg/ml, respectively. Phyllaemblicin H1 showed moderate cytotoxicity against human cancer cell lines A-549 and SMMC-7721, with IC50 values of 4.7±0.7 and 9.9±1.3 µM, respectively.

Efficacy of endoscopic therapy for gastrointestinal bleeding from Dieulafoy's lesion.

AIM: To investigate the endoscopic hemostasis for gastrointestinal bleeding due to Dieulafoy's lesion. METHODS: One hundred and seven patients with gastrointestinal bleeding due to Dieulafoy's lesion were treated with three endoscopic hemostasis methods: aethoxysklerol injection (46 cases), endoscopic hemoclip hemostasis (31 cases), and a combination of hemoclip hemostasis with aethoxysklerol injection (30 cases). RESULTS: The rates of successful hemostasis using the three methods were 71.7% (33/46), 77.4% (24/31) and 96.7% (29/30), respectively, with significant differences between the methods (P < 0.05). Among those who had unsuccessful treatment with aethoxysklerol injection, 13 were treated with hemoclip hemostasis and 4 underwent surgical operation; 9 cases were successful in the injection therapy. Among the cases with unsuccessful treatment with hemoclip hemostasis, 7 were treated with injection of aethoxysklerol and 3 cases underwent surgical operation; 4 cases were successful in the treatment with hemoclip hemostasis. Only 1 case had unsuccessful treatment with a combined therapy of hemoclip hemostasis and aethoxysklerol injection, and surgery was then performed. No serious complications of perforation occurred in the patients whose bleeding was treated with the endoscopic hemostasis, and no re-bleeding was found during a 1-year follow-up. CONCLUSION: The combined therapy of hemoclip hemostasis with aethoxysklerol injection is the most effective method for gastrointestinal bleeding due to Dieulafoy's lesion.

[Critical care and therapy based different illness state of 80 patients with severe hand-foot-and-mouth disease seen in Shenzhen].

OBJECTIVE: To discuss the treatment strategy of severe hand-foot-and-mouth disease (HFMD) cases, prevent the severe cases from progressing to fatal condition and enhance the survival rate of critically ill patients. METHODS: Eighty HFMD cases were divided into four groups, A, B, C and D, according to the severity of patients' nervous system manifestation and other system involved. Different intensive care and treatments were used and the effect and outcome were analyzed for each group. All statistical analyses were performed by using SPSS software 13.0. One-way ANOVA and Chi-square test were used for data analysis. RESULTS: The most common symptoms were continuous fever (69/70) and myoclonic jerk (67/70). The fewer the rashes were, the more severe the patient's condition was, heart rate >200/min, hypertension, increase of white blood cells in peripheral blood and hyperglycemia were common in patients with lesions in brain stem and pulmonary edema. There were no relations between patient's conditions and CSF white blood cells and CRP. CNS involvement was highly associated with EV71 infection. There were 69 cases in group A, B and C in total and all recovered. Of 11 patients in group D, 6 got complicated neurogenic pulmonary edema and circulatory failure, 2 cases died and 9 cases survived, 8 cases recovered without sequelae while one case had sequelae of mental retardation and dyscinesia. CONCLUSION: Administration of mannitol, methylprednisolone, IVIG and other supportive treatments in time and reasonably might have advantages in avoiding aggravation of the condition and enhancing the rate of successful rescue in patients with nervous system involvement.

[Changes of immune function in patients with enterovirus 71 infection].

OBJECTIVE: To investigate the association of changes in immune function with enterovirus 71 (EV71) cases with different severity of the disease. METHOD: Forty-six EV71-infected patients and 12 age-matched healthy children were enrolled in this study. The patients were divided into four groups according to critical degree of enterovirus 71 infection: hand-foot-and-mouth disease (HFMD); central nervous system disease (CNSD); autonomic nervous system dysregulation (ANSD) and pulmonary edema (PE). We analyzed CD14+ monocyte HLA-DR expression, lymphocyte immunophenotypes, the proportion of CD4+CD25+ Foxp3high regulatory T cells (Treg cells) and Th17 cells, cytokines (IL-1beta, TNF-alpha, IL-10, TGF-beta, IL-6, IL-17A), evaluated the mRNA levels of Foxp3 and ROR-gammat, and serum immunoglobulin and complements. RESULT: (1) Serum concentrations of IL-1beta and TNF-alpha elevated in mild cases, while declined in severe cases, and were lower in PE group (P<0.05). Serum concentrations of IL-10 and IL-10/TNF-alpha ratio gradually raised with the aggravation of the disease, and higher in PE group (P<0.05). (2) Circulating CD14+ monocyte HLA-DR expression, CD3+T cells, CD4+T cells, CD8+T cells, and NK cells gradually decreased, and lower in PE group (P<0.05). There was no significant difference in B cells, immunoglobulin and complement among the four groups. (3) The proportion of CD4+CD25+ Foxp3high Treg cells, mRNA level of Foxp, and serum concentrations of TGF-beta gradually decreased with the aggravation of the disease, while the proportion of Th17 cells, serum concentrations of IL-17A, mRNA level of ROR-gammat, and IL-6 gradually increased with the aggravation. CONCLUSION: Immune function changed with different illness phases. The mild cases presented systemic inflammatory response syndrome status, while critically ill cases presented compensatory anti-inflammatory response syndrome or mixed antagonist response status. Immunoregulatory treatment of patients with EV71 infection should emphasize different methods at different stage and individualization.