Application of molecular markers in the research of genetic diversity in medical helminths / 中国热带医学

@#Human-animal parasitic diseases caused by medical helminths are hazardous to human health. Genetic polymorphism studies on medical helminth populations can not only understand the biological characteristics and genetic structure of their populations, but also help reveal how they adapt to their parasitic environment, thus contributing to deepen our understanding of the epidemiological patterns of parasitic diseases and improve our understanding of accurate prevention and control of parasitic diseases. With the development of molecular biology, molecular markers such as DNA barcodes, simple sequence repeats, and single nucleotide polymorphism markers have been widely used to study the genetic relationships among parasite populations and individuals, and to reveal the genetic variation of parasite populations and the evolution of species origins. In this paper, we systematically review the application of three molecular markers commonly used in the study of genetic polymorphism in medical helminths, with a view to laying the foundation for related research.

Nucleosides-based identification model for Fritillariae Cirrhosae Bulbus / 中国中药杂志

A high performance liquid chromatography( HPLC) method was established for the fast,and precise determination of ten nucleosides in Fritillariae Cirrhosae Bulbus and its counterfeits. Then multivariate statistical analyses,such as clustering analysis,principal component analysis( PCA),and Fisher' s linear discriminant analysis( LDA),were conducted to establish a discriminant function model for an integrated analysis. The results indicated that data acquisition time of a single sample was shortened within 16 min by the HPLC method. In the range of 5-1 000 mg·kg~(-1),the mass concentrations of all nucleosides exhibited good linear relationships with the corresponding peak areas( R2> 0. 999). The spiked recoveries were in the range of 93. 83%-108. 9% with RSDs of0. 12%-1. 3%( n = 5). The limit of quantitation( LOQ) was 0. 98-4. 13 mg·kg~(-1). As revealed by the clustering analysis,Fritillariae Cirrhosae Bulbus and the counterfeits could be discriminated into two clusters based on the content of nucleosides. Fisher's LDA could achieve this discrimination,while PCA dimension reduction failed. The accuracy of the discriminant function model established on the screened characteristic indicators reached 97. 5%. The present study proposed a new identification method of Fritillariae Cirrhosae Bulbus with one-dimensional indicators,which is simple,accurate,and reliable. It can provide a scientific basis for further optimizing the identification techniques for Fritillariae Cirrhosae Bulbus and inspiration for quality control strategy development of Chinese medicinal materials.

Performance of loop-mediated isothermal amplification (LAMP) assay for detection of Schistosoma japonicum infection in Oncomelania snails in schistosomiasis transmission-interrupted regions / 中国血吸虫病防治杂志

Objective To compare the effectiveness of loop-mediated isothermal amplification (LAMP) assay and microscopic examinations for detection of Schistosoma japonicum infections in Oncomelania hupensis in transmission-interrupted regions, so as to provide insights into the optimization of snail surveillance tools in these regions. Methods Four hilly schistosomiasis-endemic villages where transmission interruption was achieved were selected in Heqing County of Yunnan Province as the study villages, including Xinzhuang and Gule villages in hilly regions and Lianyi and Yitou villages in dam regions. Snail survey was performed by means of systematic sampling combined with environmental sampling in July 2018. All captured snails were identified for S. japonicum infections using microscopy. In addition, 10 to 20 snails were randomly sampled from each snail habitat following microscopy, numbered according to environments and subjected to LAMP assay. The positive rate of settings with S. japonicum-infected snails was compared among villages. Results A total of 7 949 living snails were captured from 83 snail habitats in 4 villages, and no S. japonicum infection was detected in snails. There were 226 mixed samples containing 1 786 snails subjected to LAMP assay, and positive LAMP assay was found in 3 mixed samples from 3 snail habitats in 2 dam villages. The positive rates of settings with S. japonicum-infected snails were comparable between Lianyi Village (one setting) and Yitou Village (2 set tings) (5.89% vs. 14.29%, P = 0.344). However, the overall positive rate of settings with S. japonicum-infected snails was significantly higher in dam villages (9.67%, 3/31) than in hilly villages (0) (P = 0.048). Conclusions LAMP assay is more sensitive to detect S. japonicum infections in O. hupensis than conventional microcopy method, which may serve as a supplementary method for detection of S. japonicum infections in O. hupensis in high-risk snail habitats in hilly transmission-interrupted regions.

Clinical research progress on disappearing colorectal liver metastases / 中华胃肠外科杂志

Colorectal cancer is currently one of the most common digestive system tumors, and the liver is the most common metastatic site of colorectal cancer. In recent years, with the continuous development of the multidisciplinary treatment for colorectal cancer patients, there are quite a few cases of disappearing liver metastases (DLM) after receiving preoperative chemotherapy (or combined targeted drug therapy), and the diagnosis and treatment of DLM is currently still a very challenging and controversial topic. This article sorts out the related researches on DLM in recent years, mainly including the following 4 aspects: (1) The factors associated with DLM, including the size and number of liver metastases, chemotherapy regimens and cycles, targeted therapy drugs, and the pattern of liver metastases, Ras/Braf status and the location of the primary lesion. (2) The relationship between DLM and true complete response (pathological complete response and persistent clinical complete response), and the related predictive factors of pathological complete response. (3) Clinical evaluation of DLM: preoperative evaluation includes ultrasound, CT, MRI, and PET, while intraoperative evaluation includes intraoperative exploration, intraoperative ultrasound, and augmented reality. (4) DLM treatment strategies, including surgical treatment, local treatment, non-surgical treatment and individualized treatment.

MicroRNA144￣3p targeting histone deacetylase 2 promoting cardiomyocyte hypertrophy and cardiac function damage / 解剖学报

Objective To investigate the role and possible mechanism of microRNA(miR)144-3p in promoting cardiomyocyte hypertrophy. Methods Forty-five C57BL/ 6 mice were divided into control group, myocardial hypertrophy model group (model group), and miR144-3p transfection group (transfection group) according to their transfection method. The cardiac function related indexes of three groups of mice were detected. HE staining was performed on mouse myocardial tissue.The expression of miR144-3p in mouse cardiomyocytes was detected by Real-time PCR. Antinuclear factor (ANF), β-myosin heavy chain (β-MHC), actin α1 (Acta1) and histone deacetylase 2 (HDAC2) were detected by Western blotting in three groups. Results Compared with the control group, the interventricular septal thickness- diastolic(IVSd), interventricular septal thickness-systolic(IVSs), diastolic left ventricular posterior wall thickness(IVPWd), systolic left ventricular posterior wall thickness(IVPWs), ejection fraction(EF), cardiac weight index and left cardiac index of the model group and the transfection group were significantly higher, while systolic left ventricular diameter (LVDs) and diastolic left ventricular diameter(LVDd)were lower (P0. 05). Compared with the control group, the relative expression of miR144-3p in the model group and the transfection group was significantly higher than that in the model group (P<0. 05). Compared with the control group, the expression levels of antinuclear factor, β-myosin heavy chain, Actinα1 and histone deacetylase 2 in the model group and the transfected group were significantly higher (P<0. 05). Conclusion miR144-3p can aggravate cardiac hypertrophy by up-regulating HDAC2 and is expected to become a new therapeutic target.

Analysis of classical prescription Jinshui Liujun Jian based on ancient literature / 中国中药杂志

To provide the ancient literary evidence support for the clinical application and development of classical prescription based on systematical collection and analysis of the ancient Chinese medical literature containing Jinshui Liujun Jian, including its origin and development. Bibliometric analysis was used and information of Jinshui Liujun Jian in ancient Chinese medical literature was then collected for statistical analysis of formula compositions, main indications, dosage, preparation methods, etc. A total of 151 valid items of data were obtained from 48 ancient Chinese medicine books. Jinshui Liujun Jian was first recorded in Jingyue Quanshu written by ZHANG Jiebin. This prescription consisted of Rehmanniae Radix Praeparata, Angelicae Sinensis Radix, Pinelliae Rhizome, Citri Reticulatae Pericarpium, Poria and Glycyrrhizae Radix et Rhizome Praeparata cum Melle, and it was mainly used to treat the deficiency of lung and kidney, edema and excess production of phlegm, or Yin deficiency in the old, insufficient blood-qi, wind-cold evil, cough and disgusting, asthma and excessive phlegm. Doctors in later dynasties mostly followed the prescription compositions, dosages and indications in Jingyue Quanshu, and extended the clinical application of this prescription.

Large- scale prospective clinical study on prophylactic intervention of COVID-19 in community population using Huoxiang Zhengqi Oral Liquid and Jinhao Jiere Granules / 中国中药杂志

To scientifically evaluate the intervention effect of Chinese medicine preventive administration(combined use of Huo-xiang Zhengqi Oral Liquid and Jinhao Jiere Granules) on community population in the case of coronavirus disease 2019(COVID-19), a large cohort, prospective, randomized, and parallel-controlled clinical study was conducted. Total 22 065 subjects were included and randomly divided into 2 groups. The non-intervention group was given health guidance only, while the traditional Chinese medicine(TCM) intervention group was given two coordinated TCM in addition to health guidance. The medical instructions were as follows. Huoxiang Zhengqi Oral Liquid: oral before meals, 10 mL/time, 2 times/day, a course of 5 days. Jinhao Jiere Granules: dissolve in boiling water and take after meals, 8 g/time, 2 times/day, a course of 5 days, followed up for 14 days, respectively. The study found that with the intake of medication, the incidence rate of TCM intervention group was basically maintained at a low and continuous stable level(0.01%-0.02%), while the non-intervention group showed an overall trend of continuous growth(0.02%-0.18%) from 3 to 14 days. No suspected or confirmed COVID-19 case occurred in either group. There were 2 cases of colds in the TCM intervention group and 26 cases in the non-intervention group. The incidence of colds in the TCM intervention group was significantly lower(P<0.05) than that in the non-intervention group. In the population of 16-60 years old, the incidence rate of non-intervention and intervention groups were 0.01% and 0.25%, respectively. The difference of colds incidence between the two groups was statistically significant(P<0.05). In the population older than 60 years old, they were 0.04% and 0.21%, respectively. The incidence of colds in the non-intervention group was higher than that in the intervention group, but not reaching statistical difference. The protection rate of TCM for the whole population was 91.8%, especially for the population of age 16-60(95.0%). It was suggested that TCM intervention(combined use of Huoxiang Zhengqi Oral Liquid and Jinhao Jiere Granules) could effectively protect community residents against respiratory diseases, such as colds, which was worthy of promotion in the community. In addition, in terms of safety, the incidence of adverse events and adverse reactions in the TCM intervention group was relatively low, which was basically consistent with the drug instructions.

Molluscicidal activity of 25% wettable powder of pyriclobenzuron sulphate against Oncomelania hupensis robertsoni / 中国血吸虫病防治杂志

Objective To evaluate the molluscicidal activity of a novel molluscicide pyriclobenzuron against Oncomelania hupensis robertsoni in the mountain regions of Yunan Province, and test its toxicity to fish, so as to provide scientific evidence for the extensive application of this molluscicide in schistosomiasis-endemic foci of Yunan Province. Methods In the laboratory and snail-breeding field of Dali Prefecture, Yunnan Province, the molluscicidal activity of 5% wettable powder of pyriclobenzuron sulphate (25% PBU) against O. hupensis robertsoni was assessed by using the immersion and spraying method, and the acute toxicity of 25% PBU to carp fries was tested, while 25% wettable powder of niclosamide ethanolamine salt (50% WPNES) served as a control. Results The 1-, 2- and 3-day 25% PBU LC50 and LC90 values were 0.47, 0.25 and 0.23 mg/L, and 1.54, 0.61 and 0.49 mg / L for O. h. robertsoni by using the immersion method in laboratory, and immersion with 25% PBU at 1.0 mg / L for 1 day achieved a comparable molluscicidal efficacy in relative to 50% WPNES at 1.0 mg/L. Spraying with 25% PBU at 4.0 g/m2 achieved 1-, 3- and 7-day snail mortalities of 64.23%, 96.67% and 100.00% in laboratory, respectively, which were not significantly different from those caused by treatment with 50% WPNES at 1.0 g/m2 (all P values > 0.05). One-day field immersion with 25% PBU at doses of 1, 2 and 4 g/m3 resulted in snail mortalities of 90.00%, 93.33% and 100.00%, respectively, which were not significantly different from those caused by treatment with 50% WPNES at 1.0 g/m3 (all P values > 0.05), and 3-day field spraying with 25% PBU at doses of 2.0 and 4.0 g/m2 caused snail mortalities of 86.36% and 87.72%, respectively, which were not significantly different from those caused by 50% WPNES treatment (both P values > 0.05). The 24-, 48- and 72-hour LC50 values of 25% PBU to carp fries were 29.38, 24.62 and 23.38 mg/L, respectively, and no fish death was observed within 72 hours of exposure to 25% PBU at a concentration of 17.5 mg/L and lower. Conclusion 25% PBU is a novel, highly potent and environment-friendly molluscicide that is feasible in fish ponds, and the recommended dose is 1 g/m3 for field immersion and 2 g/m2 for field spraying in schistosomiasis-endemic areas of Yunnan Province.

Molluscicidal activity of 25% wettable powder of pyriclobenzuron sulphate against Oncomelania hupensis robertsoni / 中国血吸虫病防治杂志

Objective To evaluate the molluscicidal activity of a novel molluscicide pyriclobenzuron against Oncomelania hupensis robertsoni in the mountain regions of Yunan Province, and test its toxicity to fish, so as to provide scientific evidence for the extensive application of this molluscicide in schistosomiasis-endemic foci of Yunan Province. Methods In the laboratory and snail-breeding field of Dali Prefecture, Yunnan Province, the molluscicidal activity of 5% wettable powder of pyriclobenzuron sulphate (25% PBU) against O. hupensis robertsoni was assessed by using the immersion and spraying method, and the acute toxicity of 25% PBU to carp fries was tested, while 25% wettable powder of niclosamide ethanolamine salt (50% WPNES) served as a control. Results The 1-, 2- and 3-day 25% PBU LC50 and LC90 values were 0.47, 0.25 and 0.23 mg/L, and 1.54, 0.61 and 0.49 mg / L for O. h. robertsoni by using the immersion method in laboratory, and immersion with 25% PBU at 1.0 mg / L for 1 day achieved a comparable molluscicidal efficacy in relative to 50% WPNES at 1.0 mg/L. Spraying with 25% PBU at 4.0 g/m2 achieved 1-, 3- and 7-day snail mortalities of 64.23%, 96.67% and 100.00% in laboratory, respectively, which were not significantly different from those caused by treatment with 50% WPNES at 1.0 g/m2 (all P values > 0.05). One-day field immersion with 25% PBU at doses of 1, 2 and 4 g/m3 resulted in snail mortalities of 90.00%, 93.33% and 100.00%, respectively, which were not significantly different from those caused by treatment with 50% WPNES at 1.0 g/m3 (all P values > 0.05), and 3-day field spraying with 25% PBU at doses of 2.0 and 4.0 g/m2 caused snail mortalities of 86.36% and 87.72%, respectively, which were not significantly different from those caused by 50% WPNES treatment (both P values > 0.05). The 24-, 48- and 72-hour LC50 values of 25% PBU to carp fries were 29.38, 24.62 and 23.38 mg/L, respectively, and no fish death was observed within 72 hours of exposure to 25% PBU at a concentration of 17.5 mg/L and lower. Conclusion 25% PBU is a novel, highly potent and environment-friendly molluscicide that is feasible in fish ponds, and the recommended dose is 1 g/m3 for field immersion and 2 g/m2 for field spraying in schistosomiasis-endemic areas of Yunnan Province.

Analysis of lipophilic components of Salvia miltiorrhiza roots and S. yunnanensis roots by UPLC and LC-MS/MS / 中国中药杂志

Fingerprints of lipophilic components in the roots of Salvia miltiorrhiza and S.yunnanensis were analyzed by UPLC-DADand UPLC coupled with mass spectroscopy to evaluate the differences and similarities of the lipophilic components in the two kinds of herbs.The UPLC analysis of 18 batches of S.miltiorrhiza and 16 batches of S.yunnanensis was performed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Shimadzu LC-20AD;mobile phase was 0.026%phosphoric acid(A)-acetonitrile(B)with gradient elution;flow rate was 0.4 m L·min~(-1);detection wavelength was set at 270 nm;injection volume was 2μL.The molecular structures of the lipophilic components were analyzed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Thermo U3000 UPLC Q Exactive Orbitrap LC-MS/MS with a mobile phaseconsisting of 0.1%formic acid water(A)and 0.1%formic acidacetonitrile(B).The mass spectrometry was acquired in positive modes using ESI.There are 10 common peaks in the lipophilic components of S.miltiorrhiza.The similarity between the 16 batches of S.miltiorrhiza and their own reference spectra was greater than 0.942,and the average similarity was 0.973.There are 12 common peaks in the lipophilic components of S.yunnanensis.The similarity between the 18 batches of S.yunnanensis and their own reference spectra was greater than 0.937,and the average similarity was 0.976.The similarity between the reference chromatograms of S.miltiorrhiza and S.yunnanensis was only 0.900.There are three lipophilic components in S.yunnanensis,which are not found in S.miltiorrhiza,and one of which isα-lapachone.There is a lipophilic component in S.miltiorrhiza not found in S.yunnanensis,which may be miltirone.The two herbs contain 8 common lipophilic components including dihydrotanshinoneⅠ,cryptotanshinone,tanshinoneⅠ,tanshinoneⅡ_A,nortanshinone in which the content of tanshinoneⅡ_A,dihydrotanshinoneⅠand cryptotanshinone of S.yunnanensisis significantly lower than that of S.miltiorrhiza(P<0.01),and the contents of tanshinoneⅠand nortanshinone are significantly lower than that of S.miltiorrhiza too(P<0.05).There are significant differences in the types and contents of lipophilic components between the roots of S.miltiorrhiza and S.yunnanensis,and the similarity between the fingerprints of interspecies is much lower than that between the same species.Therefore,the roots of S.miltiorrhiza and S.yunnanensis are two kinds of herbs which are quite different in chemical compounds and compositions.

Advances in etiology, epidemiology and genetic diversity of Thelazia callipaeda / 中国血吸虫病防治杂志

Thelazia callipaeda, T. californiensis and T. gulosa are three causative agents of human thelaziasis. Most of the reported cases were caused by T. callipaeda, occurring in the old world, particularly in Asian and European countries. T. californiensis and T. gulosa have rarely been reported infecting humans in North America. T. callipaeda has long been called the oriental eye worm, referring to its traditional distribution across eastern and southeastern Asia (i.e., China, Korea, Japan, Indonesia, Thailand, and India) where infection is endemic in animals and humans, usually in poorer rural areas and mainly among children and the elderly. The identification of the parasite was mainly based on the characteristics of oral and genital organs. In Asia and Europe, vectors for this nematode are male Phortica okadai and P. variegata drosophilids respectively, which feed on ocular secretions of hosts and transmit infective stage larvae to domestic and wild carnivores, lagomorphs, and humans. China probably has the largest number of cases with thelaziasis in the world, and lots of cases have been existed in other Asian countries such as Japan and Korea. Although a few of human cases have been reported, there were high infection rates of wild animals and domesticated dogs and cats in most of European countries. Based on the cox1 gene, a total of 21 haplotypes were identified in the samples from worldwide, in which, one circulated only in European countries (h1), while the other 20 haplotypes were distributed in Korea, Japan and China. In general, the Chinese clinical isolates of T. callipaeda expressed high genetic diversity. The population differences between Europe and Asian countries were greater than those among China, Korea and Japan. The T. callipaeda populations from Europe and Asia should be divided into two separate sub-populations. These two groups started to diverge during the middle Pleistocene.

Molluscicidal effect and mechanism of action of niclosamidate against Onco-melania hupensis / 中国血吸虫病防治杂志

Objective To evaluate the molluscicidal effect of niclosamidate against Oncomelania hupensis in laboratory and explore its mechanism by determining the enzyme activities of six important enzymes in snail soft tissues.Methods O.hupensis snails were treated with niclosamidate at the concentration of 1.25 mg/L for 24 h and the snail soft tissues were separated and pre-pared for analysis.The enzyme activities of NOS,AChE,SDH,LDH,ACP and AKP were determined by ultraviolet spectropho-tometry.The morphology of the snail soft tissue was also observed.Results Niclosamidate exhibited a potent molluscicidal ef-fect against O.hupensis at the concentration of 5.00 mg/L with a mortality of 96.67% by the immersion method in laboratory.Af-ter immersed with niclosamidate(1.25 mg/L)for 24 h,the enzyme activities of NOS,AChE,ACP and AKP were significantly decreased compared with those of the controls(all P<0.01).There were no significant changes observed in the enzyme activi-ties of SDH and LDH(both P>0.05).Conclusion Niclosamidate possesses a potent molluscicidal effect against O.hupensis and its molluscicidal mechanism is probably by affecting the transmission of neurotransmitters,interfering with the circulation, metabolism and motor functions that require NO,and hindering the digestion and absorption of nutriments,which eventually re-sult in the death of the snails.

Effects of drying temperature, growing area and plucking time on two saponins from leaves of Hedera helix / 中成药

AIM To investigate the effects of drying temperature,growing area and plucking time on hederacoside C,α-hederin from leaves of Hedera helix L..METHODS Three batches of H.helix leaves plucked in different time from two growing areas were dried in a vacuum oven to the constant weight at 60 ℃,70 ℃,80 ℃,90 ℃ and 105 ℃,respectively.Two saponins in the processed leaves were determined by HPLC.The powders of the processed H.helix leaves of different batches were mixed with proper ratios,which were determined by least squares optimization method with constraints.RESULTS The content of hederacoside C in the processed H.helix leaves of the three batches increased while that of α-hederin decreased with increasing temperature.The relative error between measured value and desired contents of hederacoside C and α-hederin in the mixed H.helix leaves was less than 5.5％.CONCLUSION The effects of three factors on the content of two saponins in the H.helix leaves are in the order of drying temperature,growing area and plucking time.Mixing processed H.helix leaves of different quality statues reasonably can control the contents of two saponins in a certain range.

Study on 4 weeks medium intensity aeraobic exercise induce the difference expression of the proteomics in rat atrial muscle / 中国应用生理学杂志

OBJECTIVE@#To investigate the effect of moderate-intensity aerobic exercise on the differential expression of rat atrial muscle Proteomics and genes, which provide research basis for the rehabilitation of chronic cardiovascular diseases and exercise -induced cardiac remodeling research.@*METHODS@#Twenty male SD rats were randomly divided into control group and experimental group (=10) according to body weight. Rats in the experimental group were trained (6 days per week),which lasted for 4 weeks of moderate-intensity aerobic exercise at a rate of 24 m·min for 40 min (load intensity equivalent to 60%~70% VO). The proteins were separated by two dimensional gel electrophoresis, and the tandem time-of- flight mass spectrometer technique was used to identify 13 candidate target protein spots. The expression levels of these 13 protein spots were up-regulated more than 5 times or down -regulated to below 1/5. The mRNA of six target proteins were detected by reverse transcription-polymerase chain reaction (RT-PCR).@*RESULTS@#By software analysis, the experimental group compared with the control group, there were 8 protein points which their expression reduced more than 4/5 and 5 protein points up-regulated more than 5 times, 13 proteins were identified by mass spectrometry protein spots, the final identification results acquired 8 proteins and a unknown protein of molecular mass 54 KDa, such as:pyruvate dehydrogenase E1α1, mitochondrial aconitate hydratase, protein disulfide isomerase A3, methylmalonic acid semialdehyde dehydrogenase, mitochondrial dihydrolipoic acid dehydrogenase, isovaleryl coenzyme A dehydrogenase, glutathione synthetase, mitogen-activated protein kinase 3 and so on. Compared with the control group, the mRNA expression of methylmalonic acid semialdehyde dehydrogenase in the atrial muscle of rats was decreased after 4 weeks of moderate aerobic exercise (0.05); The mRNA expression level of isopentenyl-CoA dehydrogenase was increased (>0.05). The results indicated that the mRNA expression level was not completely consistent with the changes in mass spectrometry identification results.@*CONCLUSIONS@#The 4 weeks moderate-intensity aerobic exercise induced ignificant changes of rats atrial muscle protemics. The majority of the 13 identified target proteins in this experiment are energy metabolism enzymes. The majority of the expression of the target protein and the mRNA expression in the atrial muscle is inconsistent and different. Exercise may affect the regulation of gene transcription or downstream translation and modification of these target proteins, resulting in the change of differential expression.

Comparative study on contents of four alkaloids in homologous herbal medicines--Arecae Pericarpium and Arecae Semen / 中国中药杂志

To establish a high performance liquid chromatography (HPLC) method for simultaneous determination of four alkaloids(arecoline， guvacoline， arecaidine， and guvacine) in Arecae Pericarpium (AP) and Arecae Semen (AS), and compare the contents of these four alkaloids between different medicinal parts. The chromatographic conditions were as follows:Welch SCX（4.6 mm×250 mm， 5 μm）column, with acetonitrile-0.2% phosphoric acid solution (adjusted to pH 3.85-3.90 with ammonium hydroxide) at 50:50 as the mobile phase, at a flow rate of 0.5 mL·min⁻¹. The column temperature was set at 35 °C, and the detection wavelength was 215 nm. The results of content determination in 7 batches of AS and 10 batches of AP showed that, the contents of 4 alkaloids in AS (arecaidine 0.020%-0.045%, guvacine 0.031%-0.086%, arecoline 0.194%-0.346%, and guvacoline 0.065%-0.094%) were generally higher than those in AP (arecaidine 0.10%-0.032%, guvacine 0.006%-0.029% arecoline 0.00%-0.070%, and guvacoline 0.00%-0.020%), and most of the APs had no arecoline and arecaidine at all in fruit peel. The above results indicated that different alkaloids can be used to distinguish the different medicinal parts of Arera catechu. Arecoline， guvacoline， arecaidine， and guvacine can be used as the quality control markers of AS, while for AP, only arecaidine and guvacine were needed.

Emergency clinical laboratory system for civilian and military uses in actual military operations / 医疗卫生装备

Objective To develop an emergency clinical laboratory system for civilian and military uses to enhance medical support of filed medical unit for mass casualties.Methods The missions of field medical unit were analyzed in the actual confrontation,and an emergency clinical laboratory system for civilian and military uses was built to fulfill clinical laboratory support.The factors were explored for clinical laboratory examination,and some measures were taken accordingly.Results The requirements of field medical unit were met by the developed system.Conclusion The system can be applied in largescale military exercises to enhance clinical laboratory support ability of field medical unit.

Exploration of rapidly determining quality of traditional Chinese medicines by (NIR) spectroscopy based on internet sharing mode / 中国中药杂志

Because of the numerous varieties of herbal species and active ingredients in the traditional Chinese medicine(TCM),the traditional methods employed could hardly satisfy the current determination requirements of TCM.The present work proposed an idea to realize rapid determination of the quality of TCM based on near infrared(NIR)spectroscopy and internet sharing mode. Low cost and portable multi-source composite spectrometer was invented by our group for in-site fast measurement of spectra of TCM samples. The database could be set up by sharing spectra and quality detection data of TCM samples among TCM enterprises based on the internet platform.A novel method called as keeping same relationship between X and Y space based on K nearest neighbors(KNN-KSR for short)was applied to predict the contents of effective compounds of the samples. In addition,a comparative study between KNN-KSR and partial least squares(PLS)was conducted. Two datasets were applied to validate above idea:one was about 58 Ginkgo Folium samples samples measured with four near-infrared spectroscopy instruments and two multi-source composite spectrometers,another one was about 80 corn samples available online measured with three NIR instruments. The results show that the KNN-KSR method could obtain more reliable outcomes without correcting spectrum.However transforming the PLS models to other instruments could hardly acquire better predictive results until spectral calibration is performed. Meanwhile,the similar analysis results of total flavonoids and total lactones of Ginkgo Folium samples are achieved on the multi-source composite spectrometers and near-infrared spectroscopy instruments,and the prediction results of KNN-KSR are better than PLS. The idea proposed in present study is in urgent need of more samples spectra, and then to be verified by more case studies.

Four-year follow-up of corneal aberrations and visual functionsof myopic patients after laser in situ keratomileusis

Objective: To report on 4-year follow-up of corneal higher-order aberrations and daily visual functions of myopic patients after laser in situ keratomileusis [LASIK]

Methods: One hundred thirty four eyes of 67 patients who underwent LASIK guided by aspherical ablation were included in this study. The vision, corneal spherical aberration [SphA] and Coma were recorded before LASIK and at 6 month and 4 year after LASIK. The evaluation of the questionnaire about daily visual functions was performed by the same physician after LASIK

Results: No eye decreased the BCVA during 4 year follow-up. The effect index and safety index were 1.08 +/- 0.16, 1.11 +/- 0.17 and 1.12 +/- 0.16, 1.13 +/- 0.14 respectively at 6 month and 4 year post-LASIK. After LASIK the corneal SphA and Coma were significantly increased, however the difference between 6 month and 4 year post-LASIK was no statistical significance. Most patients [94.3%-92.4%] felt satisfaction or high satisfaction about the ability to perform each daily visual function after LASIK. Meanwhile there was still about 7.4%-9.2% patients who complained that they could not drive at night. Further analysis showed that the score of driving at night was negative correlation with corneal SphA [r=-0.645, p=0.040; r=-0.688, p=0.040 at 6 month and 4 year post-LASIK respectively]

Conclusions: Our four-year follow-up outcomes indicated that the myopic patients after LASIK had the long-term stable corneal aberration and satisfaction of daily visual functions

The Clinical Evaluation of Perioperative Integrated Operation Approaches in Operation Treatment of Pernicious Placenta Previa / 昆明医科大学学报

Objective To investigate the clinical effect of perioperative integrated operation approaches on the operation treatment of pernicious placenta previa.Methods 50 patients with pernicious placenta were divided into the treatment group (29 cases) and the control group (21 cases).Patients in treatment group were treated by integrated operation approaches,namely before cesarean section,patients were treated by uterine artery catheterization and after the baby was delivered,interventional therapy was applied immediately;patients in the control group were treated by routine operation method.The amount 24 h bleeding,hospital stay,hysterectomy rate and neonatal asphyxia rate were compared in the two groups.Results Compared with the control group,operative time,bleeding volume,bleeding volume in 24 h,postoperative hospitalization time in treatment group were shortened obviously,hysterectomy rate was lower significantly (P＜0.05);the neonatal asphyxia rate difference between the two groups had no statistical significance (P＞0.05).Conclusion Integrated operation approaches can shorten the operation time,bleeding and reduce the hysterectomy rate for patients with pernicious placenta previa,which can maintain the physical and mental health as much as possible,so it is a safe and effective treatment and worthy of clinical promotion.

Serodiagnosis of trichinellosis by ELISA using recombinant nudix hydrolase of Trichinella spiralis

Trichinella spiralis nudix hydrolase (TsNd) gene encoding a 46 kDa protein was expressed in Escherichia coli and the potential of recombinant TsNd protein (rTsNd) as an antigen for the serodiagnosis of trichinellosis was investigated by ELISA and compared with those of ELISA with T. spiralis muscle larval excretory–secretory (ES) antigens. The sensitivity of both ELISA was 100% (30/30), for the detection of anti-Trichinella IgG antibodies in sera of the experimentally infected mice, and the specificity of rTsNd-ELISA and ES-ELISA was 100% (54/54) and 98% (53/54), respectively (P>0.05). Serum anti-Trichinella antibodies were firstly detected by rTsNd-ELISA at 14 days post infection (dpi), then continued to increase with a detection rate of 100% at 36 dpi. The anti-Trichinella antibody levels at different times after infection were statistically different (P<0.05). The results showed that the rTsNd might be a potential candidate antigen for specific serodiagnosis of trichinellosis. But, it needs to be further evaluated with sera of the patients with trichinellosis and other helminthiasis.