Protective Effect of Flos Puerariae，Hoveniae Semen， and Their Compatibility on Acute Alcoholic Gastric Mucosal Injury in Mice and Mechanism / 中国实验方剂学杂志

ObjectiveTo explore the improvement effect of Flos Puerariae， Hoveniae Semen， and their compatibility on acute alcoholic gastric mucosal injury， and lay a foundation for further development of Flos Puerariae， Hoveniae Semen， and their compatibility in the prevention and treatment of alcohol-induced multiple organ injury. MethodThe acute alcohol-induced gastric mucosal injury model of mice was established by multiple intragastric administration of 56% Hongxing Erguotou liquor （15 mL·kg-1）. A total of 120 male ICR mice were randomly divided into 8 groups， namely， the blank group， model group， omeprazole group （0.026 g·kg-1）， Flos Puerariae-Hoveniae Semen （compatibility） high， medium， and low-dose groups （29.2，14.6， 7.3 g·kg-1）， Flos Puerariae group （19.5 g·kg-1）， and Hoveniae Semen group （19.5 g·kg-1）， with 15 mice in each group. After one week of adaptive feeding， the animals were pre-administrated with the corresponding drug at the rate of 10 mL·kg-1 for 3 d. From the 4th day， after 1 h of administration， Erguotou liquid was administrated at the rate of 15 mL·kg-1 and the blank group was administrated with the same volume of deionized water to record the drunkenness and sober up time. The administration was lasted for 3 d. One hour after the last administration， the eyeballs were removed and the mice were sacrificed. The concentration of ethanol in serum was determined by gas chromatograph， and the activity of ethanol dehydrogenase （ADH） in gastric mucosa was determined by ultraviolet-vis spectrophotometer. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in gastric mucosa. Serum inflammatory factors were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of nuclear transcription factor-κB （NF-κB） p65 and NF-κB inhibitory protein α （IκBα） were detected by real-time polymerase chain reaction （Real-time PCR）. ResultAs compared with the normal group， the content of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） in serum of mice in the model group was increased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues was increased （P<0.01）， and the mRNA expression of IκBα was decreased （P<0.01）. As compared with the model group， the drunkenness time of the omeprazole group， high and medium-dose compatibility groups， and Flos Puerariae group was prolonged （P<0.05）， the sober up time of the high and medium-dose compatibility groups was shortened （P<0.05）， the ethanol concentration in the serum of the high-dose compatibility group was decreased （P<0.05）， the ADH activity in the gastric mucosa of the omeprazole group and high and medium-dose compatibility groups was increased （P<0.05）， the macroscopic injury score of the high， medium， and low-dose compatibility groups and Flos Puerariae group was decreased （P<0.05）， the score of pathological injury in the omeprazole group， high， medium， and low-dose compatibility groups， and Flos Puerariae group was decreased （P<0.01）， the expression of IL-6 in serum of all drug groups was decreased （P<0.05）， the expression of IL-1β in serum of the omeprazole group， high， medium， and low-dose Flos Puerariae groups， and Hoveniae Semen group was decreased （P<0.05）， the expression of TNF-α in serum of high and medium-dose groups was decreased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues of all drug groups was decreased （P<0.05）， and the mRNA expression of IκBα in gastric mucosa tissues of the omeprazole group and high， medium， and low-dose compatibility groups was increased （P<0.05）. As compared with the high-dose compatibility group， the drunkenness time in the low-dose compatibility group and Hoveniae Semen group was shortened （P<0.01）， the sober up time in the Flos Puerariae and Hoveniae Semen groups was prolonged （P<0.01）， the concentration of ethanol in the serum of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group increased （P<0.05）， the macroscopic injury score of the medium and low-dose compatibility groups and Hoveniae Semen group was increased （P<0.05）， the pathological injury score of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， the content of IL-1β in serum of low-dose compatibility group， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， and the mRNA expression of IκBα in gastric mucosa of the Flos Puerariae group and Hoveniae Semen group was decreased （P<0.05）. As compared with the medium-dose compatibility group， the drunkenness time in the Hoveniae Semen group was shortened （P<0.05）， the sober up time in the Flos Puerariae group was prolonged （P<0.05）， the pathological injury score in the Flos Puerariae group and Hoveniae Semen group was increased （P<0.01）， and the content of IL-1β in serum of the low-dose compatibility group， the Flos Puerariae group， and Hoveniae Semen group was increased （P<0.05）. As compared with the low-dose compatibility group， the pathological injury score of the Hoveniae Semen group was increased （P<0.05）. ConclusionFlos Puerariae， Hoveniae Semen， and their compatibility play a role in preventing and treating acute alcoholic gastric mucosal injury in mice， which may be related to the inhibition of the expression of NF-κB signal pathway in gastric mucosa， and the high-dose compatibility group has the optimal effect.

The expression and immune function of pattern recognition receptor NOD2 signal in Myd88-/- mice model of Mycobacterium infection / 中国感染与化疗杂志

Objective To investigate the expression and immune function of NOD2 signal in MyD88-/- mice. Methods MyD88-/- mice and wild-type C57 BL/6 mice were characterized by PCR. Mice model of pulmonary infection was constructed by tracheal instillation of BCG vaccine strain(attenuated strain of Mycobacterium). PBS tracheal instillation was used as negative control.Peripheral blood sample and lung tissue were collected aseptically 24 h after Mycobacterium challenge. Real-time PCR and Western blot were used to detect the expression of NOD2 gene and protein. IL-6 level in the peripheral blood was determined by enzymelinked immunosorbent assay. Results The expression of NOD2 protein in BCG infected mice was significantly higher than PBS negative control group. NOD2 protein expression in MyD88-/- mice was higher than in wild-type mice. BCG infection was associated with higher NOD2 protein expression than infection-free PBS control in both groups of animals. The IL-6 level in peripheral blood was significantly higher after BCG infection than PBS group in both MyD88-/- mice and wild type mice. Conclusions BCG can activate the NOD2 signaling pathway when MyD88-dependent pathway is deficient.

Influence of liquid nitrogen cryopreservation to survive capability of Babesia microti standard strain / 中国人兽共患病学报

We discussed the influence of liquid nitrogen cryopreservation to survive capability of Babesia microti standard strain.The whole blood of mice infected with Babesia microti was put in liquid nitrogen to cryopreservation for 1 month,3 months,6 months,9 months,the whole blood was get out respectively and recovery at room temperature,and infected 3 mice respectively,100 μL/ mouse (the first generation after redissolution,the experiment group).In the same time,3 mice were also infected with Babesia microti as the animal conservation control group.When the infection rate was at a high level,the whole blood of the experiment group mice were injected into 3 normal BALB/c mice (the second generation after redissolution),to observe the changes of the Babesia microti form and proliferation situation,and also to observe the infection rate of the first and the second generation after redissolution in different conserving time.Compared with Babesia microti of animal subcultivation,the form of Babesia microti of liquid nitrogen cryopreservation changed a little.Small trophozoites,annular trophozoites,schizont and immature and mature merozoite and other form can also be seen.Compared with Babesia microti of animal subcultivation,the first time to see the worms and the time attaining to the high infection level were 1 to 2 days later,but for the second generation after redissolution,it is the same.There was no significant difference in different conserving time of 1,3,6,9 months.The influence of liquid nitrogen cryopreservation to survive capability and worm form of Babesia microti is a little,so liquid nitrogen cryopreservation can be a better way to conserving Babesia microti.

MicroRNAs recruit eIF4E2 to repress translation of target mRNAs

MicroRNAs (miRNAs) recruit the RNA-induced silencing complex (RISC) to repress the translation of target mRNAs. While the 5' 7-methylguanosine cap of target mRNAs has been well known to be important for miRNA repression, the underlying mechanism is not clear. Here we show that TNRC6A interacts with eIF4E2, a homologue of eIF4E that can bind to the cap but cannot interact with eIF4G to initiate translation, to inhibit the translation of target mRNAs. Downregulation of eIF4E2 relieved miRNA repression of reporter expression. Moreover, eIF4E2 downregulation increased the protein levels of endogenous IMP1, PTEN and PDCD4, whose expression are repressed by endogenous miRNAs. We further provide evidence showing that miRNA enhances eIF4E2 association with the target mRNA. We propose that miRNAs recruit eIF4E2 to compete with eIF4E to repress mRNA translation.

Immune identification and mass spectrometric analysis of the soluble antigens from Babesia microti / 中华地方病学杂志

Objective To analyze the fractional proteins and immunoreactivity of the soluble antigens from Babesia microti (B.microti),and find the candidate antigens for diagnosis with high sensitivity and specificity.Methods BALB/c mice were inoculated with B.microti-infected red blood cells by intraperitoneal injection.The B.microti were collected from the infected red blood cells when the infection rate reached its peak (infection rate ＞70％),then the soluble antigens were extracted by repeated freezing-thawing and ultrasonic method.The mice sera before and after the infection with B.microti for 7,14,21,28,35,42,49 and 56 days were also collected.The polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze protein components of the soluble antigens of B.microti and the Western blot was used to analyze the immunoreactivity of the soluble antigens with the pooled mice sera before and after the infection.The specific positive protein bands were identified by Liquid chromatography electrospray ionisation tandem mass spectrometry (LC-ESI-MS/MS),and the amino acid sequences of the proteins were analyzed by bioinformatics tools.Results The results from SDS-PAGE analysis indicated that the soluble antigens of B.microti showed distinct protein bands with the range between 12 and 185 × 103 (kDa,relative molecular mass,Mr),among which 9 main bands and 12 minor bands were obtained.In the Western blot analysis,the protein bands with Mr at 40 and 45 kDa could be recognized by pooled mice sera 7 days after infection;the protein bands with Mr at 40,45,54 and 95 kDa could be recognized by pooled mice sera 14 days after infection;the protein bands with Mr at 27,40,45,54,95 and 110 kDa could be recognized by pooled mice sera 21 days after infection.While,the protein bands with Mr at 27,40,45,54,95,1 10 kDa and other weak-reactive bands were recognized by pooled mice sera 28-56 days after infection,and the reaction became stronger with the infection continued.There were 336 proteins,including surface antigen,heat shock protein 70,seroreactive antigen,Eta subunit of chaperonin containing t-complex polypeptide 1 and unnamed protein products,were identified as the components of soluble antigens after mass spectrometry and sequence analysis.Conclusion The 40,45 and 54 kDa protein components from the soluble antigens of B.microti may be ideal candidate antigens for diagnosis,andtheir potential applications in diagnosing of human babesiosis deserve further study.

Study on Preparation of Immobilized Thrombin Treated with Silk Fibroin / 中国药师

Objective: To investigate the effects of silk fibroin on the immobilization of thrombin. Methods: The immobilized thrombin was prepared using silk fibroin as the carrier and glutaraldehyde as the crosslinking agent. With activity yield as the index, the process conditions of silk fibroin immobilized thrombin were determined by an orthogonal test. Results:The optimum process con-ditions of immobilized thrombin treated with silk fibroin were as follows:the immobilization time was 6 h, the enzyme dosage was 2 400 NIH·g-1 casein, the temperature was 25℃ and pH was 7. 6. The activity recovery of immobilized thrombin was 67. 22%. Conclu-sion:Silk fibroin has the positive immobilization effect on thrombin.

Rapid Determination of Bisphenol Diglycidyl Ethers and Its Migration Regulations in Interior Coating of Food Cans / 分析化学

A rapid and simple ultra performance liquid chromatographic method for determination the specific migration of bisphenol A diglycidyl ether ( BADGE ) , bisphenol F diglycidyl ether ( BFDGE ) and their derivatives in food simulants was developed. Water, 3% acetic acid, 10% ethanol and sunflower oil were used as food simulants to simulate the specific migration of bisphenol diglycidyl ethers from interior coating of food cans after 10 days storage at 60℃. After the migration period, the aqueous food simulants were directly measured without any further purification, while the sunflower oil simulant was extracted by acetonitrile followed by cleaning up using solid phase extraction. Among the migration process, BADGE and bisphenol A (3-chloro-2-hydroxypropyl)glycidyl ether (BADGE·HCl) migrated into aqueous simulants were hydrolysed into bisphenol A bis ( 2, 3-dihydroxypropyl ) ether ( BADGE · 2H2 O ) and bisphenol A ( 3-chloro-2-hydroxypropyl) (2,3-dihydroxypropyl) ether (BADGE·H2O·HCl), respectively. However, BADGE and BADGE·HCl migrated into sunflower oil were not hydrolysed. The calibrating curves showed a good linearity from 0. 05 to 10 mg/L for all the 9 target compounds. The detection limits of the method for aqueous food simulants and sunflower oil stimulant were 5μg/L and 20μg/kg, respectively. The method was applied to the determination of bisphenol diglycidyl ethers migrated from 10 kinds of food cans which were intended to contact with food. The results indicate that BADGE and its derivatives were detected at 5 of the cans, and the specific migration of BADGE(or BADGE·2H2O)and BADGE·HCl(or BADGE·H2O·HCl)in 1 cans even exceed the responding limitation regulated in EC/1895/2005 .

Population ecology of medical shellfish and the infection rate of Angiostrongylus cantonensis in Longhai City,Fujian Province, China / 中国人兽共患病学报

The aim of the study was to investigate the population ecology of medical shellfish and the infection of An-giostrongylus cantonensis in Longhai ,Fujian Province ,China .Aquatic and terrestrial shellfish were collected in survey points according to different types of breeding grounds .Then ,lung-microscopy method was involved in the detection of the lung tis-sue in Ampullaria gigas .Other shellfishes were mashed to detect the third-stage larvae of Angiostrongylus cantonensis .Hom-ogenization and lung microscopy were compared in the detection of the larvae of A .cantonensis in Achatina snails .Factors re-lated to the environment and influence of shellfish hosts were also included .Results showed that 8 species of molluscans were found ,including Pila gigas ,Bellamya aeruginosa ,Bellamya lithophaga ,Melanoides tuberculata ,Achatina fulica ,Vag-inulus alte ,Philomycus bilineatus ,and Bradybaenasimilaris with 1 673 specimens in 27 survey points from 9 townships .The infectionratewas19.78% inaverage.TheinfectionrateinV.altewas56.63% (47/83);theinfectionratesforA.fulicaand P .gigas were 39 .32% (92/234) and 27 .14% (130/234) ,respectively .The infection rate of each survey point was closely re-lated to the distances from the residents living area .Morever ,A .cantonensis larvae were detected in M .tuberculata .Lung mi-croscopy and homogenization method detection rate was 87 .1%and 100 .0% ,respectively .The difference was statistically sig-nificant .In conclusion ,V .alte ,A . fulica and P .gigas were A . cantonensist infection dominant population . The infection rate was closely related to micro-ecological environment for all kinds of shellfish .M .tuberculata was the new host of A .can-tonensis .Lung microscopy method should not be used in the qualitative screening detection of A . f ulica infected with A .can-tonensist .

Establishment of A1E3 and B1C4 monoclonal antibody-based ELISA for de-tecting circulating antigen of Schistosoma japonicum and its preliminary ap-plication / 中国血吸虫病防治杂志

Objective To establish A1E3 and B1C4 monoclonal antibody-based ELISA for detecting circulating antigen of Schistosoma japonicum and explore its application value in the field. Methods The characteristics of A1E3 and B1C4 monoclonal antibodies were analyzed by SDS-PAGE and Western blotting. The SEA-based ELISA was used to evaluate the titers of A1E3 and B1C4. The orthogonal test was used to determine the best concentration of coating antibody B1C4 and optimal working concentra-tion of A1E3-HRP. Under the optimal conditions,the serum samples of 20 acute schistosomiasis cases,46 chronic schistosomiasis cases,and 20 control sera were tested to evaluate its detection sensitivity and specificity. Seventy-two antibody positive serum sam-ples from Jiangling County of Hubei Province were detected and compared to a commercially available ELISA kit,to evaluate the detection effects of this method. Results The results of SDS-PAGE demonstrated that the purified A1E3 and B1C4 contained a clear heavy chain with molecular weight of 88 000 and 52 000 respectively and had the same light chain with molecular weight of 20 000;while Western blotting demonstrated that A1E3 and B1C4 could be recognized by SEA and serum samples of acute schis-tosomiasis cases. The SEA-based ELISA demonstrated the titers of B1C4 and A1E3 were 1∶105 and 1∶30 000,respectively. The serum samples from all the acute cases and 86.9% of the chronic cases showed a positive reaction. All of the control sera from healthy persons gave a negative response. The positive rates of the double monoclonal antibody ELISA and commercial ELISA for detecting the circulating antigen were 45.8%and 43.1%respectively,and there was no significant difference between the results of the two methods. Conclusion A1E3 and B1C4 monoclonal antibody-based ELISA is established successfully. It exhibits a high sensitivity and specificity in detecting circulating antigen of Schistosoma japonicum.

Perioperative care to patients with extramammary Paget’s disease treated with resection of tumor of vulva expansion and flap repair / 现代临床护理

Objective To summarize perioperative points for nursing patients with extramammary Paget’s disease undergoing resection of tumor of vulva expansion and flap repair.Method Eleven patients with extramammary Paget’s disease were managed with resection of tumor of vulva expansion and flap repair,and with perioperative care as well.Results The tumors in all of the patients were removed completely and the flaps survived.All patients were discharged for hospitalization of(4.5±0.7)days.No flap infection or necrosis occurred.Conclusion The measures for nursing the patients with extramammary Paget’s disease undergoing resection of tumor of vulva expansion and flap repair may include preoperative preparation,mental care,postoperative observation of flaps, prevention of complications,health education,instruction on nutrition and formation of proper life style,which may be beneficial for the smooth manipulation of resection as well as for the postoperative rehabilitation.

Expression pattern of hsa-miR-9 and its association with BCL6 in EBV-positive and EBV-negative Burkitt lymphoma cell lines / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differential expression pattern of hsa-miR-9 between EBV-positive and -negative Burkitt lymphoma cell lines and its association with BCL-6.</p><p><b>METHODS</b>The expression of hsa-miR9 and BCL-6 mRNAs in EBV(+) Raji and EBV-Ramous cells in mRNA levels were detected using fluorescence quantitative PCR (QRT-PCR). The two cells lines were transiently transfected with hsa-mir9-inhibitor and hsa-mir9-minicsvia Oligofectamine 2000, and the changes in BCL6 expressions was detected using QRT-PCR and Western blotting. Annexin V/PI staining was used to analyze the apoptosis and morphological changes of the transfected cells.</p><p><b>RESULTS</b>The expression of Hsa-miR9 and BCL-6 was significantly higher in EBV(+) Raji cells than EBV(-) Ramous cells (P<0.01). BCL-6 mRNA and protein expression was reduced in EBV(+) Raji cells after transfection with hsa-miR9-inhibitor but up-regulated in EBV(-) Ramous cells transfected with hsa-miR9-minics. Flow cytometry revealed a significantly decreased apoptosis rate in EBV(+) Raji cells transfected with hsa-miR9-inhibitor but an increased rate in EBV(-) Ramous cells transfected with hsa-miR9-minics, and the results were confirmed by microscopic observations.</p><p><b>CONCLUSION</b>Hsa-miR9 positively regulate the expression of BCL-6 and apoptosis of EBV(+) Raji cells and EBV(-) Ramous cells.</p>

Dynamics of specific antibody and circulating antigen in serum from the mice infected with Armillifer agkistrodontis / 中国人兽共患病学报

In this study,adults of Armillifer agkistrodontis (A.agkistrodontis) were collected from Agkistrodon acutus,and then the eggs were separated to feed mice.In the next step,when the infection model was established,blood serum of infected mice were collected after 1,2 and 3 weeks,respectively.Furthermore,ELISA and dot- ELISA were used to detect the dynamic change of specific antibodies and circulating antigens respectively.The specific antibodies increased from 8th week,reached the top at 12th week,decreased from 16th week,and then maintain at the same level constantly.Meanwhile,the specific antibodies were typed.It is evident that IgM antibody appeared first.However,it was substitute by IgG1 after 16 weeks.Moreover,the circulating antigens have been detected in the 1st week by dot-ELISA.Then,the dilution between 1:8 to 1:128were founded in 3rd week.The highest dilution with 1:256 appeared at 8th week,maintained before 11th week and then decreased gradually,which might provide a significant clinical implication for early diagnosis of circulating antigens.

Modern research on compatibility of Ginseng with Trogopterus Dung / 中华中医药杂志

The‘eighteen clashes’and‘nineteen fears’have been tought of traditional prohibited combination in long time,but many examples show ginseng and trogopterus dung could be safely used after compatibility,so many scholars fouce on the essential of compatibility.While people can not reach an accordant consideration for compatibility of Ginseng with Trogopterus Dung,therefore we have a overview based on documents for about 30 years,which will provide evidences for researching systematically on the nature of compatibility of Ginseng with Trogopterus Dung.

X-ray diagnosis of salpingian diverticulum--a report of 7 cases / 华中科技大学学报（医学）（英德文版）

Seven cases of typical salpingian diverticulum were identified by hysterosalpinography (HSG). The differentiation diagnosis of the disease was discussed. HSG is believed to be the method of choice for the diagnosis of this disease.

X-ray diagnosis of salpingian diverticulum--a report of 7 cases / 华中科技大学学报（医学）（英德文版）

Seven cases of typical salpingian diverticulum were identified by hysterosalpinography (HSG). The differentiation diagnosis of the disease was discussed. HSG is believed to be the method of choice for the diagnosis of this disease.

Observation on Decline Phase of Maintenance of Brugia malayiMeriones unguiculatus Model / 中国寄生虫学与寄生虫病杂志

Objective To observe the degeneration of Brugia malayi in Meriones unguiculatus model. Methods Microfilaria of Brugia malayi derived from Meriones unguiculatus was used to infect Anopheles sinensis . Infective stage larvae (L 3) from mosquito vector were collected and inoculated into abdomen of Meriones unguiculatus. Successive 33 generations of the parasite in the rodent model have been observed. Results Since 1974 when the animal model was established, the parasite has lasted for 33 generations, the positive rate of Meriones unguiculatus with microfilaria gradually reduced from 80% of the 28th generation to 16% of the 32nd generation and finally to 0 at the 33rd generation. Conclusion It became difficult for the larvae of Brugia malayi to develop and/or reproduce in the animal model after multiple inoculations for generations.