Phylogenetic and mutation point analysis of DNA barcoding sequences in Polygonatum cyrtonema / 中草药

Objective: In this work, phylogenetic analysis was used to compare the ITS2 and psbA-trnH sequences of Polygonatum cyrtonema samples from different geographical sources, so as to explore the genetic diversity and genetic relationship of these resources. Methods: PCR method was used to amplify the regions of ITS and psbA-trnH, and the sequences of ITS2 and psbA-trnH were obtained after the amplified fragment sequences were blasted in NCBI database. The neighbor joining (NJ) and maximum parsimony (MP) methods were used to construct phylogenetic trees and Kimura two-parameter (K2-P) model was used to calculate the genetic distance of different samples. Mega and DNAman softwares were applied for mutiple alignment of ITS2 and psbA-trnH sequences of 25 samples of P. cyrtonema. Results: The lengths of ITS2 and psbA-trnH sequences of Anhui Qingyang and Fujian Taining samples of P. cyrtonema were 224 bp and 620 bp, respectively. The lengths of ITS2 and psbA-trnH of the remaining 24 samples were 225 bp and 621 bp, respectively. ITS2 and psbA-trnH had seven and four mutation points, respectively. These 25 samples were clustered into two groups based on ITS2 sequences. Five samples in Hunan and Guizhou were clustered into one group, while the other 20 samples were clustered into another group. The genetic distance showed that the samples from Huaxi and Jianhe in Guizhou Province and Jianyang in Fujian Province had the largest genetic distance. Phylogenetic tree constructed by psbA-trnH sequences were unable to distinguish 25 samples from different geographical sources. Conclusion: Phylogenetic and mutation analysis will provide the theoretic foundation to utilize the resources of P. cyrtonema, investigate their evolution, and evaluate their genuineness. The results of mutation point will also be used in the identification of related P. cyrtonema resources.

Cultivable bacterial diversity of Paris polyphylla var. chinensis with 16 S rRNA sequence analysis / 中草药

Objective To analyze the bacterial diversity in stem rot and healthy plants of Paris polyphylla var. chinensis. Methods Bacterial strains were isolated from rhizomes, stems and leaves of the diseased and healthy plants of P. polyphylla var. chinensis using beef extract-peptone medium. Using 16 S rRNA universal primers 27F/1492R for PCR amplification, combined with DNA sequencing technology to preliminary identification the bacterial strains. Results The bacteria could be divided into 23 microbial species, belonging to 11 genera, isolated from healthy and diseased plants of P. polyphylla var. chinensis. Eleven endophytic bacteria, belonging to 4 genera were contained from healthy plants, in which there were 9, 10, and 5 species isolated from rhizomes, stems and leaves, respectively. Fourteen endophytic bacteria, belonging to 10 genera were contained from diseased plant, in which there were 11, 8, and 3 species isolated from rhizomes, stems, and leaves, respectively. The content of endophytic bacteria in rhizome of healthy plant was the highest, reached up to 2.999 × 105 cfu/g, while that in leaf was the lowest with 7.32 × 104 cfu/g. The quantities of Bacillus species in rhizome (73.3%), stem (67.1%), and leaf (81.8%) of healthy plant were the highest groups, Pseudomonas species in rhizome (35.6%), stem (50.3%) and leaf (60.5%) of diseased plants were the highest groups. Shannon-Wiener index and evenness index of healthy plant of P. polyphylla var. chinensis were higher than that of diseased plants. Conclusion The dominant group in the healthy plant of P. polyphylla var. chinensis was Bacillus, while the dominant groups in the diseased plant of P. polyphylla var. chinensis was Pseudomonas. The population diversity of the cultivable bacteria in healthy plant of P. polyphylla var. chinensis was more abundant than that of diseased plant.

Frequency, distribution of CD4+, CD8+ T cells and expression of CD38 in gastric mucosa of HIV infections / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the frequencies and distribution of CD4+ T cells and CD8+ T cells as well as the changes of immune activation in gastric mucosa of HIV-infected individuals.</p><p><b>METHODS</b>42 HIV-infected individuals were recruited into this investigation, and 36 patients had definite diagnosis of clinical stage. Biopsy of gastric mucosal tissues was performed by fiberoptic gastroscope including 10 normal people as a control group. Then, immunohistochemistry was used to detect expression of CD4, CD8 and CD38 in gastric mucosa, and the distinctions among three groups were analyzed with LEICA Qwin image analysis system.</p><p><b>RESULTS</b>(1) Compared with asymptomatic HIV carriers and control group, CD4 T cells remarkably decreased in the gastric mucosa of AIDS patients (P < 0.01). In gastric mucosa of asymptomatic HIV carriers, there were still some CD4+ T cells in lymphoid follicles and stroma where CD4+ T cells were unevenly distributed, the frequency of CD4+ T cells was not significantly different between asymptomatic HIV carriers and control group (P > 0.05); (2) Phenomenon of CD8+ T cells infiltrating mucosal epithelium and gland was general in HIV-infected individuals. CD8+ T cells took on local excessive hyperplasia in gastric mucosa of some individuals. As compared with control group, CD8+ T cells markedly increased in gastric mucosa of infected individuals (P < 0.01), but the distinction of asymptomatic HIV carriers and AIDS patients was not significant (P > 0.05); (3) CD38-expressing cells mainly distributed over gastric mucosal surface to superficial layer(1/3-2/3 layer) of HIV-infected individuals, and was more intensive than control group (P < or = 0.01), but there was not noticeable difference between asymptomatic HIV carriers and AIDS patients (P > 0.05).</p><p><b>CONCLUSION</b>The frequencies and distribution of gastric mucosal CD4+ T cells of HIV-infected individuals were closely correlated with progression of disease. Disfunction of mucosal immune system which was resulted from HIV infection and injury of CD4+ T cells could be an important cause of CD8+ T cells increasing and CD38-expressing enhancement.</p>

Primary application of identification-aided system for diagnosis and differential diagnosis of emerging infectious diseases / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To verify the rate of diagnostic fitting between the clinic and the indentification-aided for diagnosis and differential diagnosis system, for emerging infections diseases (EID) established.</p><p><b>METHODS</b>314 cases of 49 kinds of contagious diseases diagnosed and another 186 patients with fever who not diagnosed were tested by the system.</p><p><b>RESULTS</b>Preliminary verification was made in 314 cases diagnosed which classified to 49 kinds of contagious diseases of infectious diseases and the results showed that the coincidence rate of clinical diagnosis and first diagnosis of this system was 61.9%; the suggestive rate of first three diagnoses was 78.1%, and that of first five diagnoses was 86.6%. The diagnosis of another 186 patients with fever were diagnosed by the system and the results showed that the coincidence rate of clinical diagnosis and first diagnosis was 59.7%; the suggestive rate of first three diagnoses was 77.9%, and that of first five diagnoses was 85.4%.</p><p><b>CONCLUSIONS</b>The system can accurately suggest impossible diagnosis and differential diagnosis, and be useful for our medical work.</p>

Randomized double-blinded and controlled clinical trial on treatment of HIV/AIDS by Zhongyan-4 / 中国结合医学杂志

<p><b>OBJECTIVE</b>To assess the efficacy and safety of Zhongyan-4 (ZY-4, a Chinese herbal preparation worked out according to the therapeutic principle of supplementing qi, nourishing Yin, clearing heat and detoxication) in treating HIV/AIDS patients in the early or middle stage.</p><p><b>METHODS</b>Adopted was randomized double-blinded and placebo-parallel-controlled method, with 72 HIV/AIDS patients randomly divided into the ZY-4 group (36 patients) treated with ZY-4 and the control group (36 patients) treated with placebo. The treatment course was six months. The index of CD(4)(+), CD(8)(+) counts, body weight, clinical symptom scoring were estimated at 4 time points (0, 1, 3 and 6 month in the course), and also the viral load before and after treatment. The whole course of observation was completed in 63 patients, 30 in the ZY-4 group and 33 in the control group.</p><p><b>RESULTS</b>CD(4)(+) count in the ZY-4 group got elevated by 7.70 +/- 150.96/mm(3) on average, while that in the control group lowered by 27.33 +/- 85.28/mm(3). Fifteen out of the 30 patients in the ZY-4 group had their CD(4)(+) count increased, which was evidently much higher than that in the control group (8/33, P < 0.05), suggesting that the efficacy of ZY-4 is superior to that of placebo in elevating CD(4)(+) count. Moreover, ZY-4 showed actions in elevating CD(45)RA(+) and CD(8)(+) count, reducing HIV virus load, improving clinical symptom/sign and increasing body weight of patients. No obvious adverse reaction was found in the clinical trial.</p><p><b>CONCLUSION</b>ZY-4 has an immunity-protective and/or rebuilding function in HIV/AIDS patients in the early and middle stage, and also shows effects in lowering viral load, increasing body weight and improving symptoms and signs to a certain degree.</p>

Changes of blood cells in patients with severe acute respiratory syndrome / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the changes of blood corpuscles of patients with severe acute respiratory syndrome (SARS) in Beijing.</p><p><b>METHODS</b>Totally 43 patients (21 male and 22 female, 19-74 years old age range) diagnosed as of probable SARS were included in this study. Their corpuscles in the peripheral blood were tested every two days, and the results were analyzed.</p><p><b>RESULTS</b>Patients with SARS were more likely to develop leukocytopenia, lymphopenia and thrombocytopenia in the early period of disease than those in control group. The situation, especially lymphopenia and thrombocytopenia, could not be reversed in patients who died. Persistent low counts of lymphocytes and platelets at presentation might be associated with adverse outcomes.</p><p><b>CONCLUSION</b>Low counts of leukocytes, lymphocytes and platelets were common among patients in the early stage of SARS. Persistent Lymphopenia and thrombocytopenia may be associated with the prognosis.</p>