Correlation between changes in flavor compounds and microbial community ecological succession in the liquid fermentation of rice wine.

Microorganisms play an important role in regulating flavor compounds in rice wine, whereas we often don't understand how did they affect flavor compounds. Here, the relations between flavor compounds and microbial community ecological succession were investigated by monitoring flavor compounds and microbial community throughout the fermentation stage of rice wine. The composition of microbial community showed a dynamic change, but 13 dominant bacterial genera and 4 dominant fungal genera were detected throughout the fermentation stages. Saccharomyces presented a strong negative correlation with fungi genera but had positive associations with bacteria genera. Similarly, flavor compounds in rice wine were also showed the dynamic change, and 112 volatile compounds and 17 free amino acids were identified in the whole stages. The alcohol-ester ratio was decreased in the LTF stage, indicating that low temperature boosts ester formation. The potential correlation between flavor compounds and microbial community indicated that Delftia, Chryseobacterium, Rhizopus and Wickerhamomyces were the core functional microorganisms in rice wine. These findings clarified the correlation between changes in flavor compounds and in microbial community in the liquid fermentation of rice wine, and these results have some reference value for the quality improvement and technological optimization in liquid fermentation of rice wine.

An enzymatic colorimetric whole-cell biosensor for high-throughput identification of lysine overproducers.

L-lysine is a crucial nutrient for both humans and animals, and its main commercial use is as a supplement in animal feed to promote chicken and other animal growth. Fluorescence biosensors based on the transcriptional regulator have been developed for high-throughput screening of L-lysine producers. However, due to its inability to specifically detect lysine, this fluorescent biosensor cannot be employed to screen high-yielding strains. Here, we present a novel technique for observing L-lysine concentrations within individual Corynebacterium glutamicum cells. The transcriptional regulator LysG and its binding site, as well as the phytoene desaturase that catalyzes the synthesis of the red pigment, make up the functional core of the biosensor. The lysine-sensitive mutant LysG(E123Y, E125A), which improved the sensitivity of biosensors, was generated by site-directed saturation mutagenesis. In addition, we increased the lysine-induced chromogenic biosensor response to 320 mM by optimizing the L-lysine export mechanism and the pathway for the synthesis of lycopene precursors. The direct identification of producers with elevated L-lysine accumulation is thus made straightforward by colorimetric screening. Lys-8, a lysine producer with a maximum lysine titer of 316.2 mM, was sorted out based on the biosensor. The enzymatic colorimetric biosensor constructed here is a simple tool with great potential for the development of high-level lysine-producing C. glutamicum.

[Dynamic Changes of Ultrasonographic Images of Zusanli (ST 36) Region During Deqi of Acupuncture Needle Stimulation].

OBJECTIVE: To observe the dynamic changes of ultrosonographic images of Zusanli (ST 36) area during Deqi after inserting acupuncture needles, so as to provide objective evidence for clinical Deqi. METHODS: Sixty healthy volunteers (30 men and 30 women) were recruited in the present study. The subjects were asked to take a supine position on a test-bed, sterilized disposable filiform needles were perpendicularly inserted into bilateral ST 36 till Deqi (without lifting, thrusting and twirling), when, high-resolution ultrasound scanning was conducted over the regional skeletal muscles by using a radio-frequency (5-12 MHz) coil probe (Philips IU Elite Ultrasound Diagnostic Instrument) after smearing a layer of gel at the skin of ST 36. RESULTS: Among these 120 points of the 60 volunteer subjects, a marked Deqi was obtained from 108 points (consisting of 90%) including 52 points in men and 56 in women, and 53 at the left ST 36 and 56 at the right ST 36. Ultrosonographic scanning displayed that when a strongest Deqi feeling was obtained from bilateral ST 36 in these 60 volunteers, the acupuncture needle-tip was about (25.32±5.82) mm in the vertical depth, and was (5.45±0.55) mm lateral to the tibia, involving the anterior tibial muscle near the deep fascia in 46 acupoints, and the intersection fascia of anterior tibial muscle and extensor digitorum longus in 62 acupoints. CONCLUSIONS: The anterior tibial muscle near the deep fascia and the intersection fascia of anterior tibial muscle and extensor digitorum longus around ST 36 are involved in strongest Deqi. There are no significant differences in the gender and location (left and right).

Gastrodia elata attenuates inflammatory response by inhibiting the NF-κB pathway in rheumatoid arthritis fibroblast-like synoviocytes.

Gastrodia elata (GE), which belongs to the Orchidaceae family, was found to possess anti-inflammatory activity. However, the effect of GE on inflammatory response in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) remains largely unknown. Thus, the aim of this study was to investigate the effects of GE on tumor necrosis factor-α (TNF-α)-induced inflammatory response in RA-FLS and the underlying molecular mechanism was also explored. Our results demonstrated that GE significantly attenuated TNF-α-induced IL-6 and IL-8 production in RA-FLS. GE also inhibited TNF-α-induced MMP-3 and MMP-13 expression in RA-FLS. Furthermore, pretreatment with GE significantly attenuated TNF-α-induced the expression of p-p65 and IκBα degradation in RA-FLS. In conclusion, this study demonstrated for the first time that GE attenuated inflammatory response by inhibiting the NF-κB pathway signaling in RA-FLS. Thus, GE might have a therapeutic potential towards the treatment of RA.

[Study on method of paeonia radix alba producing and concocting integration processing].

OBJECTIVE: To study the origin pre-treating and processing integration techniques of Paeonia Radix Alba. METHOD: Different processing integration techniques were adopted and compared with traditional processing techniques to determine drying rate, aqueous extracts and peoniflori content. RESULT: Half-dry slices baked at 100 degrees C for 20 min and steamed at 100 degrees C for 10 min had the highest peoniflori contents. Half-dry slices baked at 100 degrees C for 20 min had the highest content of aqueous extracts. Products processed with conventional method and sulfur-fumigation had the lowest content of aqueous extracts. CONCLUSION: The origin processing integration techniques of Paeonia Radix Alba lose less active ingredients than conventional processing methods.

[Effect of different treatment on endophytic bacterial communities in continuous cropping of Chrysanthemum morifoliu].

To reveal the effect of rotation cropping and bacterial manure on the growth of Chrysanthemum morifolium and screen the beneficial endophytic, the diversity of endophytic and dominant genera of different treatment groups were analyzed. Four different treatments were continuous cropping, rotation, self-made organic fertilizer and commercially available fertilizer, respectively. Endophytic bacterial diversity and dominant genera in different organs were examined using Terminal Restriction Fragment Length Polymorphism (T-RFLP). The results showed that enzyme Hae III was more appropriate than enzyme Hinfl because the number of TRFs digested by enzyme Hae III was more than that of enzyme Hinfl. In comparison of diversity, the endophytic bacterial communities' diversity index in group of cropping rotation and fertilizer was higher than that of continuous cropping which indicated that the addition of exogenous microorganism in soil could increase the diversity of plant endophyte. 18 dominant species were selected, including 3 kinds of Firmicutes, 4 kinds of Actinomycetes and 11 kinds of Proteobacteria. The results of dominant species comparison showed that the number of dominant species in continuous cropping of Ch. morifolium was significantly less than that of the rotation group. Some dominant bacteria in rotation group and fertilizer group such as Arthrobacter, Streptomyces, Streptomyces, Flavobacterium and Mycobacterium were not found in the continuous cropping of Ch. mortfolium group. Dominant species of fertilizer treatment group was similar with the rotation group, and the continuous cropping group's dominant species was more abundant. It indicates that these bacteria may be able to mitigate hindrance in continuous cropping, especially the Flavobacterium which can decompose the pathogenic fungi is worthy of further attention. Compared with leaves, there are more dominant species in roots and stems. The diversity of edophytic bacterial communities in continuous cropping of Ch. morifolium stays below than that in the rotation of Ch. morifolium, and fertilizer treatment can increase the diversity of continuous cropping so that it could mitigate hindrance in continuous cropping.

Vitamin D deficiency affects the immunity against Mycobacterium tuberculosis infection in mice.

The aim of the study is to investigate the immunological changes after stimulation with bacillus Calmette-Guerin (BCG) in mice with vitamin D deficiency. After weaning, mice were divided into the vitamin D-deficient group (-D group), the normal group (N group), and the vitamin D-supplemented group (+D group). Twelve-week-old mice were intraperitoneally injected with 0.5 mg/ml BCG (≥1.0 × 10(6) CFU/mg) and maintained for 6 weeks. Spleen lymphocytes were isolated, and the percentages of CD4(+) and CD8(+) lymphocytes were determined by flow cytometry. IFN-γ levels, IL-10 levels, and the TB-PPD-specific antibody titer were determined by ELISA. The inter-group difference was analyzed using one-way ANOVA, and multiple comparisons were analyzed using the LSD test. The percentage of CD4(+) cells was 27.1 ± 0.6 in the -D group, 23.62 ± 0.42 in the N group, and 19.46 ± 0.32 in the +D group (P < 0.05). The percentage of CD8(+) lymphocytes was 12.15 ± 0.61 in the -D group, 8.7 ± 0.64 in the N group, and 7.12 ± 0.48 in the +D group (P < 0.05). The CD4(+)/CD8(+) ratio was 2.23 ± 0.15 in the -D group, 2.71 ± 0.21 in the N group, and 2.73 ± 0.31 in the +D group (P < 0.05). The plasma IFN-γ levels were 416.42 ± 16.42 pg/ml in the -D group, 325.41 ± 11.16 pg/ml in the N group, and 276.26 ± 25.32 pg/ml in the +D group (P < 0.005). The plasma IL-10 levels were 16.45 ± 1.58 pg/ml in the -D group, 24.31 ± 2.16 pg/ml in the N group, and 26.28 ± 0.42 pg/ml in the +D group (P < 0.005). The serum TB-PPD-specific antibody level was significantly higher in the -D group than in the N and +D groups. Vitamin D deficiency affects the immunity against Mycobacterium tuberculosis infection in mice.

[Study on the initial processing of Corydalis yanhusuo].

OBJECTIVE: To establish initial processing technology of Corydalis yanhusuo. METHODS: Investigated the effect of the factors such as slice method, dry method, drying temperature on the content of water-extract, ethanol-extract, effective component in Corydalis yanhusuo pieces. Compared the quality with that of the traditional initial processing samples. RESULTS: The best initial process method was: cut fresh Corydalis yanhusuo into 4 - 5 mm thick slices, dry at 70 - 80 degrees C or microwave dry. CONCLUSION: The study provides theoretical base for modifying the initial processing.

[Therapeutic efficacy of drug susceptibility test-guided individualized anti-tuberculosis chemotherapy for spinal tuberculosis].

OBJECTIVE: To investigate the efficacy of individualized anti-tuberculosis chemotherapy guided by drug susceptibility testing for spinal tuberculosis through analyses on the post-operative follow-up outcomes. METHODS: The diagnoses of spinal tuberculosis were established by clinical, radiological and histological evaluation in 132 patients who were admitted from August 2005 to January 2010, 62 patients (37 male and 25 female) with follow-up more than 12 months in this study. The average age was 33.6 years (ranging from 4 - 67 years). The infected samples were collected during surgery. After processed in a routine laboratory procedure, the samples were inoculated into vials of the BACT/ALERT 3D system. The drug susceptibility testing was performed using absolute concentration method, which included 11 first-line and second-line drugs. Four or five anti-tuberculosis drug regimen was chosen according to the results of drug susceptibility testing. All the patients were followed up a month later, and then once 3 months in the following 11 months, and subsequently at intervals of half a year. The clinical status, erythrocyte sedimentation rate (ESR), roentgenogram, MRI and 3D-CT were concerned to estimate the progress of tuberculosis. RESULTS: The culture positive rate was 45.2% (28/62). The average detection time was 42 d (ranging from 28 - 58 d). The drug susceptibility testing showed a total drug resistance level of 24.2%:12.9% for isoniazid, 4.8% for rifampicin, 3.2% for ethambutol, 9.7% for streptomycin, 6.4% for pasiniazid, 14.5% for levofloxacin, 1.6% for rifapentine. The mean follow-up period was 21 months (ranging from 12 - 44 months). According to Bridwell criteria, grade I bony fusion was obtained in all patients in 8 - 12 months. CONCLUSION: Guided by drug susceptibility testing, individualized anti-tuberculosis chemotherapy for 12 to 18 months is effective for spinal tuberculosis.

[Comparative analysis of sequence alignment of SH3GL1 gene as a disease candidate gene of adolescent idiopathic scoliosis].

OBJECTIVE: To identify whether SH3GL1 gene serves as a disease associated gene of adolescent idiopathic scoliosis (AIS). METHODS: Positioning candidate cloning: "case-sibling or case-family control design" research scheme based on family constellation was designed. Fifty-six AIS patients (15 male and 41 female, mean age 15 years old, ranged from 8 to 22 years old, Cobb angle from 25 degrees to 110 degrees , average Cobb angle of 67.5 degrees ) from November 2007 to December 2008 were recruited. In all patients, blood preparation was collected, and genome DNA was extracted. According to nucleotide sequence of gene SH3GL1, primer pair for PCR amplification, cloning, and sequencing with 10 exons as emphasis was designed. Sequence comparative analysis for exon sequencing result between sib pairs or family pairs, and that between sib pair or family pairs and NCBI (National Center for Biotechnology Information) were conducted through Vector NTI Advance 10.3 software to judge whether basic group mutation occurred or not. Amino acid sequence comparative analysis for prediction was made. RESULTS: Ten exons of the candidate gene SH3GL1 were successfully amplified and cloned in genome DNA of an AIS sib pair and family pairs, and the sequencing obtained positive results. Twelve basic group mutations were found in 10 exons of the candidate gene SH3GL1 of patients with AIS. These mutations were located in the second exon (3 mutations), the fourth exon (1 mutations), the fifth exon (4 mutations), the sixth exon (1 mutations), the eighth exon (1 mutations), and the tenth exon (2 mutations, noncoding region). If basic group in 515 of mRNA was mutated to T, termination codon(TAG) came into being and open reading frame was altered. The sequence of protein showed brachytmema protein was encoded, which could cause changes of primary structure. CONCLUSION: SH3GL1 is possibly one of the disease associated genes of AIS.

[Study on the influence on Rhizoma Corydalis by different harvest time and processing methods].

OBJECTIVE: To choose the best harvest time and initial processing method of Rhizoma Corydalis. METHODS: Taking the content of tetrahydropalmatine and dry rates as major indices, different samples from different harvest time and different processing methods were investigated by HPLC. The C18 column (250 mm x 4.6 mm, 5 microm) was used with mobile phase of methanol-0.1% phosphoric acid solution (pH = 6.0, adjusted with triethylamine) (58: 42). The mobile phase flow rate was 1.0 mL/min. The detection wavelength was 280 nm. RESULTS: The contents of tetrahydropalmatine were similar although the harvest time was different. The dry rate on May 21 was higher than the others. The content of tetrahydropalmatine was the highest by steaming and reached 0.11%. CONCLUSION: The best harvest time for Rhizoma Corydalis is around 10 days after the above-ground plant wilted, and the best initial processing method for Rhizoma Corydalis is steaming.

[Effect of environmental factors on growth of Huperzia serrata].

OBJECTIVE: To establish and optimize the cultivation systems for wild Huperzia serrata, through assessing the effects of environmental factors on its growth. METHOD: H. serrata was planted under natural field condition, the growth status of H. serrata under different light intensities, temperatures, relative humidity (R. H.), fertilizer levels, and so on were investigated and analyzed. RESULT: H. serrata grew nicely under following environmental conditions: temperature ranging from 18.9-26.3 degrees C, R. H. ranging from 81%-90% and 1330-3000 lx light intensity. A cultivation system, in which H. serrata was planted in the bamboo forest and did not occupy the arable land, was established through the study. CONCLUSION: Concluded that light intensity and R. H are the two factors which greatly confine the growth of H. serrata. As long as the levels of the above two factors meets the demands, H. serrata could be cultivated.