This article reviewed the researches proceeding on the contamination and detection of the foodborne pathogen noroviruses (NoVs) in fresh produce, which involved the NoVs contaminations in fresh produce, the special attachment of NoVs in fresh produce, the NoVs outbreaks associated with fresh produce and the NoVs detection in fresh produce. There had been an increase in reported infectious disease risks associated with the consumptions of fresh produce for recent 30 years. Because the NoVs, as a primary cause of viral gastroenteritis thoughout the world, were highly contagious, had a low infectious dose, and were persistent in the environment. And also the methods for NoVs detection in food had significantly developed over the last 15 years. Currently NoVs were the most common pathogen accounting for 40% of outbreaks associated with fresh produce (i. e., fruits and vegetables). Data from outbreaks investigations verified fresh produce as the high risk food products for NoVs. The fresh produce were typically eaten raw with no thermal processing, can be contaminated at any step during production and processing from faecally polluted water and fertilizers, the poor hygiene practices by food handlers and the cross-contamination. The attachment of NoVs to the fresh produce was due to the physio-chemical factors of virus protein coat, the special attachment to different fresh produce, and the possibility for internalization of NoVs. It might provide answers to why those high risk foods were more frequently implicated (i. e., lettuce and raspberries). According to the data of foodborne NoVs outbreaks which were associated with fresh produce from EU countries and the USA, the outbreaks in EU countries were mainly associated with NoVs contaminated raspberries and lettuce, while in USA which were associated with NoVs contaminated lettuce. Unfortunately, there were no NoVs detection methods for fresh produce or the data of foodborne NoVs outbreaks which were associated with fresh produce in China. That made it difficult to analyze the NoVs contamination situation in China. The heterogeneous distributions of presumably low levels of virus, which presented in contaminated fresh produce, also made it difficult to detect NoVs. To solve this problem, different sampling methods, viral elution methods and RT-qPCR methods were chosen. For example, according to the isoelectric point of NoVs particles, high pH and high ionic strength solution could be used as means for releasing NoVs. For the elution from acidic fruit, the buffer capacity and the virus recovery could be increased by the addition of tris-HCl. When analyzing pectin containing raspberries or strawberries, the viral elution usually incubated with pectinase at neutral pH to avoid from foaming jelly. In this paper, the latest ISO standard for NoV detection in food and the new approaches for NoV detection were also reviewed to provide references for domestic researches. It was necessary to establish and develop domestic methods for NoV detection in fresh produce, especially the different NoV conventional molecular detection methods with corresponding NoV extraction methods, which targeted to the different adsorption characteristics of different fruits and vegetables, in order to strengthen the national food safety monitoring.
Food Analysis ; methods ; Food Contamination ; analysis ; Foodborne Diseases ; virology ; Fruit ; virology ; Gastroenteritis ; virology ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; physiology ; Vegetables ; virology

BACKGROUND:It is accepted that the best method for spermatogonial stem cells separation is using artificial cryptorchism model combined with surface makers.OBJECTIVE:To explore the feasibility of separation spermatogonial stem cells with ?6-integrin and c-kit as specific surface markers.DESIGN,TIME AND SETTING:The randomized control experiment was performed at the Renmin Hospital of Wuhan University from May to December 2006.MATERIALS:Forty adult,white Kunming mice with 6 weeks old were randomly divided into cryptorchidism and control groups,with 20 animals in each group.METHODS:Artificial cryptorchidism model was prepared by made an incision at the median of abdomen,and testis was pulled into abdominal cavity,which was fixed at the each side of lateral abdominal wall.There was no treatment in the control group.The single cell suspension of seminiferous epithelium was obtained by traditional two step enzyme digestion at 2-3 months after operation.FITC-conjugated anti-?6-intergrin antibody and PE-conjugated anti-c-kit antibodies were added.Then the cells with low side scatter light-scattering properties were sorted and positively stained for ?6-intergrin and negative c-kit expression.Meanwhile,the viability of the isolated cells was assessed by trypan blue staining.MAIN OUTCOME MEASURES:The morphological changes of cryptorchidism,and the sorting results of spermatogonial stem cells.RESULTS:Cell distribution in seminiferous tubule was disorder with reduced numbers.The layer and lumens were disappeared,and cell division phase could be seen in the center of tubules.Compared to the control group,the testicular cells in the cryptorchidism group were increased in the side scatterlow,Forward scatterhi areas,with figure left-upward displacement.The distribution of ?6-integrin+ and c-kit cells were deviated each other,it named that most ?6-integrin+ cell were not spermatogonial stem cells,so do the c-kit-cells.Only 2.8% of testicular cells exhibited side scatterlow,?6-integrin+,and c-kit-,which were spermatogonial stem cells in the cryptorchidism group.And trypan blue staining showed that over 95% of them were viable.CONCLUSION:Using the two surface markers to sort spermatogonial stem cells can advance the purity of the spermatogonial stem cells in cell suspension,but the specificity is insufficient.

Objective To study the genetic diversity of germplasm resources for Potentilla discolor.Methods Twelve species of germplasm resources for P.discolor were analyzed by ISSR molecular markers.To make up the systematic diagram of genetic relationship by POPGENE 1.32 software,cluster by UPGMA method,and establish the dendrogram.Results A total of 128 ISSR bands were scored for 12 primers,among which 101 were polymorphic bands.The average percentage of polymorphic bands was 78.91%.Genetic similarity coefficient was changed from 0.179 2 to 0.632 5.By cluster analysis,the geographical distribution is mutually related to the relationship of germplasm resources for P.discolor and it was also showed some of P.discolor from the same region were in the same group which presented the rule of geographical distribution in the tested materials.Conclusion The diversity level of the different germplasm resources for P.discolor higher and the relationship of P.discolor correlates with the geographical location in some way.

OBJECTIVE:To explore how to establish good procedure,system and mode of drug management,dispensing and application,in order to provide reference for risk management and rational use of rabies vaccine. METHODS:The use of rabies vaccine and storage temperature monitoring in our hospital during 2013-2014 were analyzed statistically,and risk management and use of rabies vaccine in our hospital were analyzed,and management measures and attentions were put forward. RESULTS:The amount of vaccinum rabiel (Vero cell) and human rabies immunoglobulin in our hospital were increased in 2014,compared to 2013. The position labeled with #1 in storehouse and that of labeled with #1 and #2 in dispensing store could meet the storage condi-tion of rabies vaccine. Risk management could be carried out in fields of drug requisition,cold chain management,drug manage-ment and drug dispensing. ADR should be paid attention. CONCLUSIONS:It is of significance to develop risk management and ra-tional medication guidance of rabies vaccine.

Objective:To analyze the adverse drug reactions(ADR)and clinical application of thalidomide to provide useful reference for rational medication in clinics. Methods:The case reports and literatures from foreign countries on the clinical medication of thalidomide were analyzed and summarized. Results:The ADR of thalidomide included gastrointestinal reaction, hematotoxicity,cadiovascular toxicity,neurotoxicity,skin lesion,pulmonary embolism and so on. Its new medication methods were widely used in clinics. Conclusion:Clinicians and pharmacists should pay more attention to the medication risks and rational use of thalidomide in order to assure the safety and effectiveness of clinical drug use.

Objective To discusses the methods of the case selection and monitoring and diagnosis and treatment to selective non-operative management of closed splenic injuries .Methods The clinical data and results of 98 cases from October 1999 to Octo-ber 2011 with closed splenic injuries managed non-operative treatment were analyzed retrospectively .Results There were 90 cases (91 .8% ) conservative successfully ,5 cases(5 .1% ) transit operation because hematoma increase and bleeding continuously during therapeutic observation and 2 cases(3 .1% ) died from multiple injuries merge infection and multiple organ failure .Conclusion Most of the closed splenic injuries below level Ⅱcan be treated by non-operative management ,the effect and prognosis determined by the reasonable case selection and monitoring conditions and diagnosis and treatment .

Aim To synthesize and identify artificial antigen of podophyllotoxin for the production of podophyllotoxin polyclonal antibody. Methods The hapten was synthesized by two different chemical approaches and characterized by TLC, IR, NMR, and MS. Mixed anhydride reaction (MAR) and active ester method (AEM) were used to couple the podophyllotoxin to carrier proteins (BSA and OVA). Characterization of artificial antigens was done by using spectroscopy and electrophoresis. The anti-podophyllotoxin polyclonal antibodies were obtained through immunizing rabbits. Results The results from IR, NMR and MS showed that 4-O-succinoyl podophyllotoxin (hapten) was successfully synthesized. The coupling molar ratios of the hapten and carrier proteins were 88.6 for Hapten-BSA1, 40.3 for Hapten-BSA2, 17.8 for Hapten-OVA1, and 54.2 for Hapten-OVA2. Hapten conjugates coupled with BSA yielded two sets of the specific and affinitive polyclonal antibodies. One set of antibodies showed an IC50 value of 2.21 μg·mL -1 with a detection limit of 0.12 μg·mL -1. Conclusion Antigenic conjugates were artificially synthesized, and based on these artificial antigens, polyclonal antibodies against podophyllotoxin were raised from rabbits immunized with two different immunogens and characterized with an indirect ELISA format.

The objective of this study is to generate broad spectrum monoclonal antibody ( mAb ) against a group of pyrethroid insecticides and to identify its immunological characteristics. The generic hapten 3-phenoxy-benzoic acid ( PBA) was conjugated to carrier protein BSA by activated ester method. Balb/c mice were immunized with PBA-BSA. The titer of polyclonal antibody ( pAb ) was detected by indirect enzyme-linked immunosorbent assay ( ELISA) after five times immunization. The mouse with high titer and sensitivity was selected for cell fusing. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by indirect non-competitive ELISA based on the coating antigen PBA-ovoalbumin ( PBA-OVA ) . High-sensitivity and high-specificity mAb was prepared after subcloning using limiting dilution method. Purified mAb was obtained after purified by saturated ammonia sulfate precipitation and protein G affinity column. The immunological characteristics of mAb such as titer, antibody subtypes, affinity constant and the sensitivity to pyrethroid insecticides were characterized by indirect ELISA; The results of UV spectroscopy and SDS-PAGE showed that PBA-BSA artificial antigen was synthesized successfully. A hybridoma cell line (4H11) secreting anti-pyrethroid mAb was established. The titre of ascites was up to 1:6. 5×106, and the mAb was IgG1 subtype. The affinity constant of the mAb to PBA was about 2. 5×107 L/mol, with a IC50 value of 208. 83 μg/L and a detection limit of 21. 23 μg/L to PBA. Simultaneously, beta-cypermethrin, flucythrinate, cypermethrin and fenvalerate were sensitively recognized by the mAb with the IC50 of 1. 01, 2. 15, 3. 16 and 3. 67μg/L, respectively.

Objective To study the endogenous glucocorticoid on rat femoral head microenvironment of 11 hydroxysteroid dehydrogenase expression ,and to discuss the influence of combined with femoral head pathological changes of the corresponding mechanism .Methods Sixty SD rats were divided into control group ,1‐month group ,3‐months group ,each 20 rats in group .1‐month group and 3‐months group inject cortisone acetate in the abdominal cavity intraperitoneal for 1‐month or 3‐months each .Im‐munohistochemical ,immunofluorescence ,Real‐time qPCR ,HE staining were employed in this study .Results From immunohisto‐chemical ,immunofluorescence ,Real‐time qPCR ,the 11 hydroxysteroid dehydrogenase content of 1‐month group and 3‐months group were higher than that of the control group(P<0 .05) .From HE staining we detected 1‐month group in the bone marrow cavity in‐creased in fat cells ,3‐months group subchondral trabecular bone density decreased ,compared with the control group(P< 0 .05) . Conclusion Supplement of corticosterone could promote rat femoral head microenvironment 11 hydroxysteroid dehydrogenase ex‐pression and subchondral trabecular bone density decrease .

ObjectiveTo investigate the diagnosis and treatment of adrenocorticotropin-independent macronodular adrenal hyperplasia (AIMAH). MethodsThe clinical data of 14 cases of AIMAH from August 1972 to July 2010 were retrospectively analyzed.The cases included 5 males and 9 females with a mean age of 45 (range 26 to 58 ) years.Ten patients demonstrated typical Cushing's syndrome (CS) and 4 patients presented with weight gain,hypertension or diabetes mellitus without any signs of CS.The circadian rhythm of serum cortisol was abnormal.Low and high dose dexamethasone suppression tests failed to suppress cortisol secretion.CT scan showed bilateral enlargement of the adrenal glands with multiple macronodules.All patients underwent open surgery,including 5 cases of unilateral adrenalectomy,6 cases of adrenalectomy combined with contralateral subtotal adrenalectomy and 3 cases of bilateral adrenalectomy.ResultsIt was established by pathological examination that all patients had bilateral adrenal macronodular or adenomatoid hyperplasia.During the mean follow-up of 69 months (range 12 to 120 months),the clinical symptoms of CS disappeared after surgery in all cases.The 5 patients who received unilateral adrenalectomy had urinary free cortisol and serum cortisol within normal ranges and no further enlargement of the contralateral gland was noticed.Among the 3 patients who received bilateral adrenalectomy,1 case died of adrenal crisis on day seven post-operation.The remaining 2 cases presented with adrenal insufficiency but returned to normal after glucocorticoid replacement therapy.Nelson's syndrome was not observed in the other patients.ConclusionsAIMAH has unique endocrinological and pathological features,presenting as an independent etiology of CS.Unilateral adrenalectomy appears to be an effective and safe alternative treatment for AIMAH and long-term remission can be achieved.Bilateral adrenalectomy or unilateral adrenalectomy combined with contralateral subtotal adrenalectomy may be performed if the symptoms have not improved or recurred after unilateral adrenalectomy.