Effects of drought stress on physiological and biochemical and chemical components of Cinnamomum cassia seedlings / 中国中药杂志

Six month old Cinnamomum cassia seedlings were used to simulate drought stress with polyethylene glycol(PEG 6000). The physiological indicators(osmotic substances, antioxidant enzymes, etc.) and chemical components of seedlings under different drought levels and the correlation between the two were studied. The results showed that the chlorophyll content and relative water content decreased gradually with the increase of PGE 6000(0, 5%, 10%, 15%) concentration and time(3, 5, 7 d), while the soluble protein content, soluble sugar content and catalase(CAT) activity increased, but the rising rate slowed down with the time. The activities of peroxidase(POD), superoxide dismutase(SOD), malondialdehyde(MDA) and proline content increased at first and then decreased. The content of coumarin, cinnamaldehyde, cinnamic acid and dimethoxycinnamaldehyde decreased, while the content of cinnamyl alcohol continued to increase.Under drought stress, the fluorescence signals of reactive oxygen species and no contents in roots of C. cassia seedlings were significantly stronger than those of the control.Further correlation analysis showed that coumarin content, di-methoxycinnamaldehyde content and osmoregulation substance content were significantly negatively correlated(P<0.05), cinnamic acid content was significantly negatively correlated with POD and SOD activities(P<0.01).It was found that C. cassia seedlings showed a certain degree of drought tolerance under short-term or mild drought stress, but if the drought exceeded a certain degree, the physiological metabolism of the seedlings would be unbalanced.

Identification of Citrus medica var. sarcodactylis Infected with Citrus Huanglongbing / 中国实验方剂学杂志

Objective::To analyze and compare different samples in many aspects to identify Citrus medica var. sarcodactylis infected with Huanglongbing(HLB) timely and accurately, in order to prevent and control the disease in time. Method::HLB was identified through character analysis, reverse transcription-polymerase chain reaction(RT-PCR), enzyme digestion reaction and Real-time PCR. Result::In terms of characters, there were typically variegated yellow leaves and relatively small fruit, even with deformity but without " red nose fruit" among C. medica var. sarcodactylis infected with HLB. All of these can be used as the basis for the preliminary identification of HLB in the fields. According to the RT-PCR test results and enzyme digestion reaction, when the primer was OI1/OI2c, there was specific band of 1 160 bp, which could be cut into 520 bp and 640 bp by Xba I enzyme. These results were consistent with the characters of other citrus plants infected with HLB. According to the Real-time PCR detection results, C. medica var. sarcodactylis infected with HLB had amplification curves and dissolved peaks, with the melting temperature was 82 ℃ and Ct between 24.6 to 28.2, while the normal plants were not amplified. Conclusion::Character analysis can be used to roughly distinguish HLB in the fields, but with a certain subjectivity. RT-PCR or Real-time PCR can be used to identify C. medica var.sarcodactylis infected with HLB in a timely and accurate manner, and qPCR detection is more sensitive and quantitative. Through the combination of character analysis and molecular identification, C. medica var.sarcodactylis infected with HLB can be determined more timely and accurately.

Advances in quality research of Cinnamomum cassia / 中国中药杂志

Cinnamomum cassis is one of the commonly used traditional Chinese medicines in China. Its genuine producing areas distribute in Guangdong and Guangxi provinces. As an important edible herb and export variety of China, the quality control and internationalization of quality standards of C. cassis is extremely significant. In the recent years, with the development of the cinnamon industry, relevant academic research and the upgrade of the international standards, it is necessary to summarize the quality-related progress of C. cassis. In the present review, the germplasm resources, specific quality marker(Q-marker) and quality standards of C. cassis were summarized on the basis of published research during the last 10 years.

Chemical Compositions and Pharmacological Activities of Polygoni Multiflori Radix / 中国实验方剂学杂志

As a common tonic traditional Chinese medical herb,Polygoni Multiflori Radix has been extensively applied in clinic. In recent years,there have been many literatures related to Polygoni Multiflori Radix. By reviewing the literatures in domestic and foreign,the latest progress on chemical components and pharmacology of Polygoni Multiflori Radix have been summarized and analyzed. At present,more than 133 kinds of compounds have been isolated from Polygoni Multiflori Radix, such as stilbene glycosides,terpenoids,flavonoids,phospholipids and phenylpropanoids. Among them, stilbene dimer and dianthrone glycosides are two kind of new compounds recently isolated from it. The current researches about the pharmacological effect of Polygoni Multiflori Radix focus on anti-oxidative,anti-tumor,anti-atherosclerosis and neuro-protective effects,with potentials in treating neurodegenerative diseases, preventing and treating arterial and reducing blood sugar. Its medicinal ingredients mainly include stilbene glycosides,terpenoids and flavonoids,in which 2,3,5,4'-tetrahydroxysilbene 2-O-glucopyranoside shows a variety of biological activities. However,there are a few studies on the pharmacological activities of other compounds. Therefore,to ensure the further development and utilization of Polygoni Multiflori Radix,it is suggested to conduct a more in-depth and comprehensive research on the material basis of its efficacy.

Methodological study for detecting gene mutation of family with genotyping of compound heterogenicity of SEA alpha-thalassemia 1 and HbCS / 中国实验血液学杂志

This study was aimed to establish a method of PCR combination with PCR-RFLP for detecting the South-East Asian (SEA) deletion type alpha-thalassemia 1 and non-deletion mutation of Hb Constant Spring (CS), and to investigate the application value of this method. For the members of the families with alpha-thalassemia, SEA deletion mutation was detected by PCR, then the HbCS point mutation was screened by PCR-RFLP. The results indicated that 15 carriers with alpha-thalassemia (--(SEA)/) were found in 19 members from 7 families, and 2 families with genotype of --(SEA)/alpha(CS)alpha were screened out successfully. It is concluded that the PCR combination with PCR-RFLP is a simple, rapid, and reliable method for screening HbH disease with genotype of --(SEA)/alpha(CS)alpha.

Method for detection of gene mutation of hemoglobin E in population of Sichuan area / 临床检验杂志

Objective To establish a PCR-RFLP method for detection of CD26 mutation of hemoglobin E(HbE)gene.Methods The genome DNA from the family members who were suspected of having HbE combined with beta-thalassemia was amplified using PCR.The PCR products were digested by restriction endonuclease Mnl I,and then separated on PAGE.The electrophoretical patterns were finally analyzed to confirm if CD26 mutation was present or not.Results Four cases with CD26 mutation in two families were successfully screened out.Conclusion Although the incident frequency of CD26 mutation in Sichuan population is not high,it must not be neglected since it may present alone or combined with other type of thalassemia.PCR-RFLP method described in this study is available in screening mutated HbE gene and treating the disease owing to its simplicity,rapidity and specificity.

Determination of sperm creatine kinase and its isozyme in infertile patients / 中国组织工程研究

BACKGROUND: As a biochemical marker of cytoplasm, the increased activity of creatine kinase (CK) in human spermatozoa is correlated with both the residual cytoplasma and the ratio of sperm with abnormal func tion. It is a marker of mature sperm and associated with the potential of in semination. OBJECTIVE: To investigate the differences of CK activity in sperm and the relative contents of sperm CK-MM and CK-BB isoenzymes between normal fertile males and infertile males, and evaluate its significance in clinical diagnosis of male infertility. DESIGN: Case-control observation. SETTING: Laboratory of Biochemistry, Family Planning Research Insti tute of Sichuan Province. PARTICIPANTS: Ninety-four male infertile patients between January 1999 and October 2000 were selected from the Department of Family Planning Sciences of Sichuan Province, who had no aspermatism with their wives proved to be fertile. The average age of subjects were 31 years. Eighty subjects with the sperm count ＞ 2×1010 L-1 were taken as normal sperm group and 14 subjects with the sperm counts ＜ 2×1010 L-1 were considered as oligospermia group. Semen obtained from 27 healthy males who were normal in routine examinations and with children was taken as the healthy control group.METHODS: Semen sample collected by masturbation after abstinence of 3 to 5 days was incubated at 37 ℃ for liquefication and routinely analyzed.Total activity of CK in sperm was determined by using a kinetic spectrophotometry and the relative contents of CK isozyme was determined by agarose gel electrophoresis followed by density scanning of CK isozyme.MAIN OUTCOME MEASURES: Sperm counts, percentages of viability and motility of sperm, total CK activity and the relative contents of CK-MM and CK-BB isozyme in spermatozoa.RESULTS: A total of 94 enrolled patients and 27 normal controls were involved in the analysis of results. ①Sperm counts, percentage of viability and motility in oligospermia group ( Ⅱ + Ⅲ, WHO method) were obviously lower than those in the healthy control group, and those in the normal sperm group, except the sperm counts, were remarkably lower than the healthy control group [the sperm counts in healthy control group, normal sperm group and oligospermia group was (6.05 ±0.81 )×1010 L-1, (7.76±1.37)×1010 L-1 and (1.46±0.19)×1010 L-1 respectively (P ＜ 0.01). The survival rate in healthy control group, normal sperm group and oligospermia group was 85.1%,56.8% and 58.2%, P ＜ 0.01, and the sperm motility was 62.9% ,34.6% and 29.5% respectively, P ＜ 0.01].②Total activity of sperm CK in oligospermia group was significantly higher than the healthy control group [respectively (9.000±6.117) and (1.933 ±0.943) kat/108 sperm,P＜ 0.05],although that in the normal sperm group (2.800±0.862) kat/108 sperm was a little higher than the healthy control group, while there were no significant difference between the two groups (P ＞ 0.05).③The relative contents of sperm CK-MM isozyme in the normal sperm group and oligospermia group were obviously decreased (that in the healthy control group, normal sperm group and oligospermia group was 30.5%, 19.0% and 18.0% respectively,P ＜ 0.05), which implied a remarkable difference in sperm differentiation between healthy control group and the latter two groups.CONCLUSION: The determination of sperm CK is meaningful for the diagnosis and pathogenesis of oligospermia. More work should be done on the distribution of sperm CK-MM isozyme in different infertile population as well as its importance in the diagnosis of infertility.