The instability of secondary metabolite production is a ubiquitous problem in plant cell culture. To understand the instability, the investigation of anthocyanin accumulation in suspension cultures of Vitis vinifera, as a model system, has been initiated in our laboratory. Suspension culture of a relatively homogeneous cell line E of V. vinifera, was established by long-term cell line selection by anthocyanin content differentiation. The aggregate size of E was smaller than that of other cell lines obtained by routine screening method. The variation coefficients of anthocyanin content in suspension cultures of E were 8.7% in long-term subcultures and 5% in repeated flasks, respectively. The effects of elicitor, precursor feeding and light irridiation on biomass and anthocyanin accumulation in suspension cultures of E had been investigated and the results showed that all the variation coefficients were lower than 12% and this indicated the importance of homogeneity on stable production in plant cell culture. With the combination treatment of 30micromol/L phenylalanine and 218micromol/L methyl jasmonate in the dark in suspension cultures of E, the anthocyanin content and production in suspension culture of E was 5.89-fold and 4.30-fold of the controls, respectively, and all the variation coefficients of biomass and anthocyanin accumulation were lower than those of the controls in 5 successive subcultures.
Anthocyanins ; biosynthesis ; Biomass ; Cell Proliferation ; Light ; Suspensions ; Vitis ; cytology ; metabolism

OBJECTIVETo analysis the reasons of fixation failure for intertrochanteric fractures, so as to select correct operation indications and fixation methods.

METHODSRetrospective analysis the clinical data of 13 patients with failed internal fixation of intertrochanteric fractures from September 1997 to September 2008, and the failure reasons were summarized. There were 7 males and 6 females,ranging in age from 58 to 93 years,averaged 71 years. Two patients were treated with intramedullary fixation, 4 patients with anatomical proximal femoral plate, 3 patients with DHS fixation, 2 patients with hollow compression screws, and 2 patients with external fixation. According to Evans types: 1 patient was type II, 7 patients were type III, and 5 patients were type IV.

RESULTSEight patients with unstable fractures and malreduction had no grafted bone. Six patients had bad position of neck screws in the femur neck. Postoperative collodiaphyseal angle: 3 patients were under 90 degree, 7 patients 90 to 110 degree, and 3 patients 110 to 130 degree. Five patients had internal fixed screw exited, 6 patients had neck screws cutting to superior lateral, 3 patients had early weight bearing, and 10 patients were osteoporosis occurred after operation from 6 weeks to 11 months, averaged 4.5 months.

CONCLUSIONThe fixation failure of intertrochanteric fractures was concerned with fractures types, reduction, fixation methods, osteoporosis and the time of weight bearing.

Aged ; Aged, 80 and over ; Female ; Fracture Fixation, Internal ; methods ; Hip Fractures ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Radiography ; Retrospective Studies ; Treatment Outcome

The low-production is a ubiquitous problem and has prevented the commercialization of secondary metabolite production in plant cell culture. In order to examine the effective approaches to improvement of secondary metabolite production in plant cell culture, the investigation of anthocyanins accumulation in suspension cultures of Vitis vinifera, as a model system, had been initiated in our laboratory. In this present research, various elicitors and the precursor of phenylalanine were used in combination to enhance the anthocyanins production in suspension cultures of Vitis vinifera. And an integrated process with the combination of elicitation, precursor feeding and light irradiation was reported for rational bioprocess design. Among the combination treatment of phenylalanine feeding and several elicitors (methyl-beta-cyclodextrin, dextran T-40, methyl jasmonate, extracts of Aspergillus niger and Fusarium orthoceras), the combination with methyl jasmonate gave the highest anthocyanins production in suspension cultures of Vitis vinifera. When compared to the controls, the anthocyanins content (CV/g, FCW) and production (CV/L) increased by 2.7-fold and 3.4-fold, respectively. The optimum time for the addition of phenylalanine and methyl jasmonate was 4 days after inoculation. Two cell lines with different anthocyanins-producing capacity responded differently to the optimum combination treatment of 30 micromol/L phenylalanine feeding, 218 micromol/L methyl jasmonate elicitation and 3000 to approximately 4000 1x light illumination. The high-and low-anthocyanins-producing cell lines of VV05 and VV06 produced the maximum of 2975 and 4090 CV/L of anthocyanins that were 2.5- and 5.2-fold of the controls, respectively.
Acetates ; pharmacology ; Anthocyanins ; biosynthesis ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Cyclopentanes ; pharmacology ; Light ; Oxylipins ; pharmacology ; Phenylalanine ; pharmacology ; Vitis ; cytology ; metabolism

The commercial application of plant cell cultures is often hindered by the instability of secondary metabolite biosynthesis, where the metabolite yield fluctuates and decline dramatically over subcultures. This study proposed that such instability is due to the fluctuations of culture variables. To validate this hypothesis, the effects of the fluctuations of two culture variables (subculture cycle and inoculum size) on the biomass, anthocyanin biosynthesig, intracellular carbon, nitrogen and phosphate during continuous 10 subculture cycles were investigated. The subculture cycle was fluctuated for 12h in a 7 day cycle (6.5, 7 and 7.5 d), and the inoculum size was fluctuated by 20% on basis of 2.00 g (1.60, 2.00 and 2.40 g). It was found that all the measured culture parameters fluctuated over the 10 subculture cycles. The fluctuation in terms of inoculum sizes had a greater effect on the stability of anthocyanin biosynthesis in suspension cultures of V. vinifera. Among all the subculture conditions investigated, 7d-subculture cycle and 1.60 g-inoculum size was the best one to hold the relatively stable anthocyanin production. The anthocyanin yield presented a negative correlation with intracellular sucrose content or intracellular total phosphate content.
Anthocyanins ; biosynthesis ; Carbohydrate Metabolism ; Cell Culture Techniques ; methods ; Intracellular Space ; metabolism ; Phosphates ; metabolism ; Plant Proteins ; metabolism ; Suspensions ; Vitis ; cytology ; growth & development ; metabolism

OBJECTIVETo observe the effect of bone marrow mesenchymal stem cell transplantation on postangioplasty aortic restenosis in rats.

METHODS48 SD rats were randomly divided into normal control group, balloon injury group, balloon injury and MSCs transplantation group. MSCs were pre-labeled by DAPI (25 microg/ml) and then infused into aorta through the balloon catheter (MSCs 2 x 10(6)/animal). Thoracic aorta were taken for histological examination (frozen and paraffin sections) at 1, 2, 6 weeks post angioplasty, respectively. DAPI labeled MSCs were detected under immunofluorescence microscopy. Expressions of c-kit, proliferating cell nuclear antigen (PCNA), smooth muscle alpha-actin (alpha-SMA) in aorta were determined by immunocytochemistry using related antibodies.

RESULTSThe DAPI-labeled MSCs could be detected on impaired intimae and alpha-SMA expression was seen in these cells 1 weeks after MSCs transplantation. Similar weak c-kit expression in neointima was found in both injury and transplantation group at 2 weeks (P > 0.05). Expressions of PCNA and alpha-SMA in the neointima were significantly higher in transplantation group than in injury group at 2 weeks. Intima/tunica media area ratio and luminal stenosis ratio were significantly increased in transplantation group than injury group at 6 weeks (all P < 0.05).

CONCLUSIONBone marrow MSCs transplanted post aortic angioplasty could home to serious wounded aortic intima, differentiate into smooth muscle like cells, promote neointima cellular proliferation and aggravate postangioplasty aortic restenosis in rats.

Angioplasty, Balloon ; adverse effects ; Animals ; Aorta ; pathology ; Bone Marrow Transplantation ; adverse effects ; Coronary Restenosis ; etiology ; pathology ; Disease Models, Animal ; Male ; Mesenchymal Stem Cell Transplantation ; adverse effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo assess the efficiency of eluting stent coated with arsenic trioxide (As(2)O(3)) suspended in poly-L-lactic acid (PLLA) to prevent in-stent restenosis in rabbits.

METHODSForty-five male New Zealand white rabbits were assigned to three groups (n = 15 for each group) at random: uncoated stents, stents coated with PLLA or stents coated with As(2)O(3) in PLLA. Animals were euthanized 28 days after stent implantation into the iliac arteries of rabbits. Neointimal thicknesses and apoptosis of vascular smooth muscle cell (VSMC) were measured. Stents coated with As(2)O(3) in PLLA were implanted in another 48 male New Zealand white rabbits, As(2)O(3) concentrations in serum and arterial tissue at implantation site were measured at 2 h and 1, 3, 7, 14, 28 days after As(2)O(3) eluting stent implantation (n = 8 for each time point).

RESULTSNeointimal hyperplasia was significantly reduced 51% and 31% and apoptosis significantly increased (21.0 +/- 3.3; 6.2 +/- 1.9(*); 5.3 +/- 2.1(*), (*)P < 0.01 vs. As(2)O(3) eluting stent) with As(2)O(3) eluting stent, versus PLLA-coated stents and uncoated stents. As(2)O(3) concentrations in arterial tissue at implantation site were 18.6 +/- 9.1 (ng/mg) at 1 day and 0.3 +/- 0.1 (ng/mg) at 28 days after stent implantation.

CONCLUSIONSAs(2)O(3) coated stents released As(2)O(3) to local tissue for at least 28 days, suppressed neointimal hyperplasia in rabbit iliac arteries and increased local VSMC apoptosis might be one of the mechanisms for inhibiting restenosis by As(2)O(3) coated stents.

Animals ; Apoptosis ; drug effects ; Arsenicals ; administration & dosage ; Coronary Restenosis ; prevention & control ; Drug-Eluting Stents ; Iliac Artery ; Male ; Muscle, Smooth, Vascular ; cytology ; Oxides ; administration & dosage ; Rabbits ; Random Allocation

OBJECTIVETo investigate the clinical features, treatment strategy and risk factors affecting the prognosis of elderly patients with non-small cell lung cancer (NSCLC) complicated by chronic obstructive pulmonary disease (COPD).

METHODSWe retrospectively analyzed the data of elderly patietns (>60 years) with newly diagnosed NSCLC complicated by COPD at the Geriatric Institution of General Hospital of PLA between January, 2000 and June, 2015. The clinical data collected included history of smoking, pulmonary function test results, initial treatments, TNM stage, chief complaints, comorbidities and laboratory tests. The Cox proportional hazards regression model was used to explore the prognostic factors in these patients.

RESULTSA total of 200 NSCLC patients were reviewed, of which 107 (53.5%) patients had the co-morbidity of COPD as confirmed by spirometry using bronchodilator test. The median survival of the patients with NSCLC complicated by COPD was 45.8 months with 1-, 3-, 5-, and 10-year survival rates of 80.4%, 55.4%, 41.0% and 20.0%, respectively. Stratification analysis showed that patients with COPD Gold grades 1 and 2 had a significant longer median overall survival (51.7 and 43.1 months, respectively) than those with grade 3/4 (16.9 months; P=0.020 and 0.043, respectively). Univariate and multivariate analyses using Cox proportional hazards regression model showed that an older age, a higher Gold grade, advanced disease stage (stages III and IV), squamous cell carcinoma, nonsurgical initial treatment, coughing and an elevated serum CEA level were independent risk factors for shorter survival of the patients.

CONCLUSIONMultiple prognostic factors can affect the outcomes of elderly patients with NSCLC complicated by COPD, and a higher COPD Gold grade that fails to respond to treatment within 3 months is the independent risk factor for survival of the patients.

This study was aimed to understand the current status of the antimicrobial resistance and molecular distribution of Campylobacter in various poultry in Jiaodong area,to provide a basis for effective prevention and control of the Campy-lobacter risk to poultry products and human health.Campylobacter was isolated and identified from 565 cloacal samples collect-ed in the Jiaodong area from August to October 2021 through conventional bacterial isolation and culture,mass spectrometry,microbroth dilution and multilocus sequence typing(MLST).The drug resistance and molecular typing of 131 representative strains(67 Campylobacter jejuni and 64 Campylobacter coli)were studied separately.Antimicrobial resistance analysis indica-ted that 131 isolates were highly resistant to ciprofloxacin,nalixic acid and tetracycline,with resistance rates of 96.21％,96.21％and 95.45％,respectively.Except for 2 strains,62 strains of C.coli were completely resistant to these three drugs(100％).A total of 65 strains of 131 strains were multidrug re-sistant,and the overall multidrug resistance rate was 49.62％,among which 11 strains(16.42％)of C.jejuni were resistance to 3-5 antibiotics,and 54 strains(84.38％)of C.coli were re-sistance to 3-6 antibiotics.Among the isolates from different poultry sources,waterfowl isolates were the most resistant,fol-lowed by broiler isolates.The MLST typing results revealed 72 alleles and 35 sequence types obtained from 67 strains of C.je-juni,and the distribution was relatively dispersed,without a dominant ST type and homologous complex.A total of 27 alleles and 19 sequence types were obtained from 64 strains of C.coli.Moreover,59.38％(38/64)strains were homologous complex CC-828,in which the ST-1586 sequence type was most frequent,followed by ST-825.ST-1586,ST-9944 and ST-3735 were the main sources of C.coli in broilers,and ST-825 and ST-1586 were the main sources of C.coli in waterfowl.Differences in C.jejuni and C.coli carriage were observed among poultry in the Jiaodong area.Carriage of the two bacteria was more common in laying hens than in broilers and waterfowl.C.jejuni from poultry in the Jiaodong area was highly resistant to ciprofloxacin,nalixic acid and tetracycline,but had good sensitivity to other drugs.C.coli was highly resistant to a variety of antibiotics,and multiple drug resistance was common.St-type dispersal of C.jejuni showed high genetic diversity.C.coli was cloned and transmitted mainly by ST-1586 in broiler chickens and waterfowl.Poultry carry C.jejuni,which can cause serious diseases in humans.Therefore,dynamic monitoring of Campylobacter from poultry should be strengthened.

PAMAM dendrimer is one of the most widely studied dendrimers in recent years, which has a large number of functional groups on the surface and cavities inside, specific three-dimensional structure and good biocompatibility, permeability and stability. It has been widely applied in drug and gene carrier fields and may become a new absorption enhancer. In order to study the absorption enhancing effects of PAMAM dendrimers, liquiritin was selected as the model drug, with the protection of spleen and liver, detoxification and other functions, but it had not been widely used in clinical application because of its difficult absorption, first pass effect, and low bioavailability. This topic was based on the two main determinants (solubility and permeability) of intestinal absorption in the body, researched the physicochemical properties of liquiritin, analyzed the transport volume of liquiritin with or without PAMAM dendrimers by using Caco-2 cell model, and analyzed the cytotoxicity of PAMAM dendrimers on Caco-2 cells by MTT experiments. These results showed that 0.1% of the G4 generation PAG can promote the absorption of liquiritin safely and effectively, and it was suitable for further development into a new type of pharmaceutical excipients.

We established a multiplex direct PCR for rapid detection of E.coli,Salmonella,Staphylococcus aureus,Listeria and Yersinia enterocolitica bacteria.Multiplex direct PCR primers were designed according to gene sequences of phoA (E.coli),inv A (Salmonella),nuc (S.aureus),hl y (Listeria),and ail (Y.enterocolitica).After the multiplex direct PCR were established,the specificity and sensitivity of primers were detected.Then,multiplex direct PCR was applied to examine 60 swine product samples,the detection specificity,accuracy and positive predictive value were calculated compared with the gold standard culture method.Results showed that multiplex direct PCR primers could be used for specific detection of E.coli,Salmonella,S.aureus,Listeria and Y.enterocolitica,with the minimal detectable limit of 10,1,100,1 and 1 CFU,respectively.For the examination of 60 swine product samples using multiplex direct PCR,15 were positive for E.coli,6 positive for Salmonella,21 positive for S.aureus,20 positive for Listeria,and 35 positive for Y.enterocolitica,with all positive detection rates higher than that of culture.The total detection sensitivity was 100％,accuracy was 94％,and positive predictive value was 81.44％.Multiplex direct PCR could be used for specific and sensitive detection of common food-borne pathogens,and the testing time was shorten to be 3 hours because of saving time for template extraction.Multiplex direct PCR might serve the detection of food-borne pathogens in food safety risk monitoring much better.