[Applications of immobilization techniques in the screening of active constituents from traditional Chinese medicines].

It has been an active approach to screen the active ingredients in traditional Chinese medicines (TCMs) according to the affinity property between small molecule compounds and biomaterials such as cells, bacteria and proteins. On the other hand, the biomaterials can be immobilized on a solid support before the screening procedure. The immobilization method not only can maintain the biological activities of biomaterials, but also have other advantages such as high efficiency, simple operation, easy to be continuous and automatic, etc. Carrier materials (solid supports) for the immobilization including silica gel, magnetic materials, hollow fiber, and the surface plasma resonance sensor chips have been used to immobilize biomaterials and successfully applied in the screening of active ingredients from TCMs. In this paper, applications of immobilization techniques in the screening of active components from TCMs were reviewed to provide a scientific reference to the future applications.

[Efficiency of multi-round fluorescence in situ hybridization and its influencing factors in preimplantation genetic diagnosis].

OBJECTIVE: To investigate the efficiency of multi-round fluorescence in situ hybridization (FISH) and its influencing factors in preimplantation genetic diagnosis (PGD). METHODS: A total of 48 couples accepted PGD because of various reasons: 24 with Robertsonian translocations, 16 with reciprocal translocations, 2 with pericentric inversions, one with advanced maternal age who had a previous liveborn of Down syndrome, 3 suffered from sex chromosome abnormalities and 2 repeated spontaneous miscarriages. After 72 retrieval cycles, 432 cleavage stage embryos with more than six cells were biopsied on day three. Only intact nuclei (396) were hybridized in order to verify the chromosomal status of the individual embryos. If previous FISH has failed to give conclusive results while the nuclei remained undamaged, the nuclei were hybridized once again. A total of 870 times of hybridization were conducted to 396 nuclei. Signal identification rates of each round as well as the influence of different probes to the hybridization efficiency were compared. Factors leading to inconclusive FISH results were analyzed as well. RESULTS: Five hundred and thirty five out of 870 hybridizations gave identifiable signals (61.5%). The second and third round FISH showed the best signals with an identification rate of 71.8% and 77.4%, respectively, which were significantly higher than those of the first round (52.8%, P < 0.01), the fourth round (55.8%, P < 0.05, P < 0.01), the fifth round (54.5%, P < 0.05) and the sixth round (27.3%, P < 0.01). The identification rate of centromere specific probe signals (CEP group) was 80.3% and the former three rounds in this group got the best quality of signals with an identification rate of 85.7%, 85.1% and 88.0%, respectively, which was significantly higher than that of the latter three rounds. The identification rate of other probe was much lower than with the CEP probe (55.2% vs. 80.3%, P < 0.01) and the best quality of signal in this group was achieved in the fifth round (72.7%), followed by the second round (66.1%) and the third round (63.8%). The identification rate of the first round (50.3%) and the sixth round (22.2%) were significantly lower compared with the second round (P < 0.01). During the 6 rounds of FISH, 335 hybridizations did not give conclusion results (38.5%, 335/870). The main cause of unidentification was weak signals (20.9%, 182/870). Other common factors included background interference (7.6%, 66/870) and failed hybridization (6.1%, 53/870). Rare causes included nucleus damage (1.8%, 16/870), nucleus loss (1.1%, 10/870) and signal split/overlap (0.9%, 8/870). CONCLUSION: Multi-round FISH can improve the utility of single nucleus in PGD and the former three rounds have the highest efficiency. The hybridization effect of CEP is better than other probe. Poor signal quality is the common cause of unidentification results.

[Cumulative effects of K-function in the research of point patterns]. / ç‚¹æ ¼å±€ç ”ç©¶è¿‡ç¨‹ä¸­K-函数的累积效应.

The spatial pattern of plant population is one of primary issues in ecological research. Point pattern analy-sis is considered as an important method to study the spatial pattern of plant population. Ripley's K function has been commonly used for point pattern analysis. However, the cumulative effect of Ripley's K function may lead to specific spatial pattern charcteristics. To explore how the cumulative effect of Ripley's K function affects population pattern, the data of clumped distribution, random distribution and regular distribution of Stipa grandis were simulated by R software. All data generated by R software were analyzed by Ripley's K function and the non-cumulative pairwise correlation function g(r). The results showed that for clumped distribution (or regular distribution), the cumulative effect of Ripley's K function was manifested in two aspects. On the one hand, the scale of clumped distribution (or regular distribution) was increased due to Ripley's K function. On the other hand, Ripley's K function could detect the difference of the distribution of cluster (or negative interaction range) in the sampling space, exhibiting different pattern characteristics. For random distribution, Ripley's K function had no cumulative effect. In conclusion, the combination of Ripley's K function and pairwise correlation function by collecting replicate samples could better reveal the essential characteristics of the pattern in the study of population pattern.

[Clinical features, diagnosis and treatment of 5 cases of primary pneumonic plague in Tibet in 2010].

OBJECTIVE: To explore the clinical manifestations, the feature of chest X-ray, the clinical outcome, and the clinical treatments of severe pneumonic plague. METHODS: We observed the clinical course of primary pneumonic plague in 5 patients, who infected Yersinia pestis in Tibet during September 2010, including manifestations of chest X-ray, the antibiotic therapy, respiratory support and the prognosis. RESULTS: All of the 5 patients presented with high fever, bloody sputum and difficulty breathing. The chest X-ray showed signs consistent with necrotizing inflammation with multiple lobar involvement. Mass-like lesions might coalesce, and the "white lung" sign might appear. Three out of the 5 patients presented with hypoxemia. The results of reverse indirect hemagglutination assay (RIHA) in these patients were positive on the second day of the illness onset. All of these patients recovered after antibiotic therapy and other treatments. However, the absorption of lung lesions was very slow. CONCLUSIONS: Patients infected with primary pneumonic plague presented with rapid onset high fever and hemoptysis, and the lung injury was very severe. The positive result of RIHA was useful for early diagnosis of plague. Streptomycin should be the first choice for Yersinia pestis infection, but its optimal dose needed further study. Fluoroquinolones can be used as combination with Streptomycin. Nutritional support and symptomatic treatment, as well as non-invasive or invasive mechanical ventilation when needed, were important for the management of the disease.

[Location of Dctn1 in the mouse testis and sperm and its role in spermiogenesis].

OBJECTIVE: To investigate the role of dynactin 1 (Dctn1) in the process of mouse spermiogenesis. METHODS: Western blot and indirect immunofluorescence were used to analyze the expression and location of Dctn1 in the mouse testis and spermatozoa. The highest efficiency of small interference RNA (siRNA) was verified by GC2-spd cell line in vitro and in vivo studies, respectively. Dctn1 siRNA mixed with the indicator (0.4% trypan blue) was injected into the seminiferous tubules of 3-week-old ICR mice through rete testis microinjection, and negative control siRNA injected into the control testes. The normal group included 3-week-old ICR mice that did not receive any treatment. Spermatozoa were collected from the cauda epididymis 3 weeks after siRNA injection for morphological analysis. RESULTS: Dctn1 was mainly localized in the tail of spermatozoa. After interference, the sperm tail abnormality in the Dctn1 siRNA group was (23.57 +/- 0.55)%, significantly higher than (12.35 +/- 2.29)% in the control (P < 0.01, n = 3), and it was (3.37 +/- 0.69)% in the normal group. CONCLUSION: Dctn1 plays an important role in mouse spermiogenesis, and mainly affects the formation of the tail of spermatozoa.

Identification of antioxidants in essential oil of radix Angelicae sinensis using HPLC coupled with DAD-MS and ABTS-based assay.

Radix Angelicae sinensis (named danggui in Chinese), a commonly used traditional Chinese medicine, has multiple pharmacological activities. The essential oil of danggui is usually considered to be its main active fraction. However, to date, studies on the antioxidant potentials of danggui have focused on water-soluble compounds. In this paper, the antioxidant activity of the commercial essential oil of danggui was investigated by DPPH radical scavenging assay, ABTS radical scavenging assay, and beta-carotene bleaching test. Antioxidant constituents in the essential oil were identified using HPLC coupled with DAD-MS and ABTS-based assay. The results showed that the essential oil of danggui has concentration-dependent antioxidant activity, which can be attributed to its component (coniferyl ferulate). This is the first report on the antioxidant activity of essential oil from danggui; its antioxidant potential was lower than those of positive controls, ascorbic acid and BHA.

[Genetic effects of workers occupationally exposed to mercury using three genetic end-points].

OBJECTIVE: To investigating genetic effects of workers occupationally exposed to mercury (Hg). METHODS: The peripheral lymphocytes from 20 workers exposed to mercury and 20 controls were measured with micronucleus test, comet assay, hrpt gene mutation test and TCR gene mutation test. RESULTS: The mean micronuclei rate(MNR) and mean micronucleated cells rate(MCR) in 20 workers were (5.90 +/- 0.91) per thousand and (5.30 +/- 0.81) per thousand, respectively while MNR and MCR in controls were (1.50 +/- 0.47) per thousand and (1.30 +/- 0.31) per thousand respectively, The difference of MNR and MCR between workers and controls was very significant (P < 0.01). The mean tail length (MTL) of workers and controls were (3.16 +/- 0.31) and (0.99 +/- 0.07) microm, respectively. The mean tail moment (MTM) of workers and controls were 1.63 +/- 0.22 and 0.39 +/- 0.03, respectively, There was a significant difference in MTL and MTM between workers and controls(P < 0.01). When the average mutation frequencies (Mfs-hprt) of hprt and (Mfs-TCR) of TCR of workers were compared with those of controls, there were not significant difference (P > 0.05). CONCLUSION: The results of the investigation indicated that the adverse genetic effects in workers occupationally exposed to mercury could be detected.

An ultrafiltration and high performance liquid chromatography coupled with diode array detector and mass spectrometry approach for screening and characterizing thrombin inhibitors from Rhizoma Chuanxiong.

Thrombin (THR) plays a significant role in thromboembolic diseases, direct THR inhibitors are a class of important clinical anticoagulant drugs. This study established a THR in-solution based biospecific extraction combined with ultrafiltration and high performance liquid chromatography coupled with diode array detector and mass spectrometry analysis (TUA) method to screen and identify ligands for THR in Rhizoma Chuanxiong. After evaluating the reliability of the present TUA method using positive (argatroban) and negative (adenosine, tirofiban, ticagrelor) control drugs, this method was successfully applied to detect eight potential active compounds in Rhizoma Chuanxiong. Two new THR-targeted compounds isochlorogenic acid C and senkyunolide I with high THR inhibitory activity (IC50 206.48 and 197.23µM, respectively) were identified by liquid chromatography/mass spectrometry and enzyme inhibitory activity test finally. They were reported with direct THR inhibition activity for the first time and their ligand-THR interactions were explored by in silico molecular docking research. In addition, based on the TUA screening result, four compounds gained similar structure with the two hit compounds were also investigated as promising candidates targeting THR with high binding energy (>5.0kcal/mol). These results may prove that the proposed method could effectively screen THR inhibitors in complex mixtures.

[Effect of Polysilicic Acid on the Deposition Behavior of Different Aluminum Species].

To understand the water quality problems caused by deposition and re-dissolution behaviors of residual aluminum in drinking water distribution system (DWDS), the effect of polysilicic acid on the deposition and re-dissolution behaviors of different aluminum species was investigated by using jar tests and quartz crystal microbalance, which could demonstrate the deposition behavior of different aluminum species and the physic-chemical properties of deposited layers. The results showed that the effect of polysilicic acid on the deposition behavior of different aluminum species was significant. Al0 (mainly monomeric species), Al13 and Al30 (polymeric species) could react rapidly with polysilicic acid, and some insoluble products formed. However, the stability of insoluble products from different aluminum species was distinct. The insoluble product formed by reaction of Al0 with polysilicic was relatively stable. With regard to the systems of polysilicic acid with Al13 and Al30, the dissolved aluminum concentration increased with reaction time, which indicated that the deposited product could be gradually transformed to soluble monomeric or oligomeric aluminum species. Compared with the influence of Al/Si ratio on Al0, the ratio of aluminum to silicon significantly affected the deposition and re-dissolution behavior of Al13 and Al30, and the re-dissolution rate of insoluble aluminum was the most rapid when the ratio of aluminum with silica was 0.2:20 (as mass ratio of Al to SiO2).