The cis-epoxysuccinate hydrolase (CESH) from Rhizobium strain BK-20 is the key enzyme for L(+)-tartaric acid production. To establish a highly efficient and stable production process, we first optimized the enzyme production from Rhizobium strain BK-20, and then developed an immobilized cell-culture process for sustained production of L(+)-tartaric acid. The enzyme activity of free cells reached (3 498.0 +/- 142.6) U/g, and increased by 643% after optimization. The enzyme activity of immobilized cells reached (2 817.2 +/- 226.7) U/g, under the optimal condition with sodium alginate as carrier, cell concentration at 10% (W/V) and gel concentration at 1.5% (W/V). The immobilized cells preserved high enzyme activity and normal structure after 10 repeated batches. The conversion rate of the substrate was more than 98%, indicating its excellent production stability.
Alginates ; chemistry ; Cells, Immobilized ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Hydrolases ; metabolism ; Rhizobium ; enzymology ; metabolism ; Tartrates ; metabolism

Objective: To explore the tongue epithelial cell apoptosis quantitatively to analysis tongue demonstration change mechanism of perimenopausal syndrome by Chaihu Shugan Decoction. Methods: Patients with perimenopausal syndrome 60 cases, divided into kidney yin deficiency (30 cases), deficiency of both kidney yin and yang (30 cases), 30 healthy cases in normal group. The treatment course lasted for 2 months. The flow cytometry was used to detect the tongue epithelial cell apoptosis. Results: The tongue epithelial cell apoptosis rate of perimenopausal syndrome patients was higher than that in the normal group (P

Objective:To establish the HPLC fingerprint of ethanol parts of compound Sanhuang tincture ( an anti-infective drug) using high performance liquid chromatography, and analyze the classification of the chemical components. Methods: A Phenomenex Luna C18 (2) (250 mm × 4. 6 mm, 5 μm) column was used. The mobile phase was acetonitrile-0. 05 mol·L-1 aqueous potassium di-hydrogen phosphate solution (22 :78) and the flow rate was 1. 0 ml·min-1 . The DAD detector was used and the detection wavelength was 237 nm. The column temperature was 30 ℃ and the injection volume was 10 μl. Results:The fingerprints of compound Sanhuang tincture were obtained with promising separation degree and the number of theoretical plates. A total of six fingerprint characteristic peaks were identified, and the reproducibility, stability and precision of the method were good. Meanwhile, combined with the informa-tion of retention time of compound medicine, single drug and reference substance, the source of characteristic peak of the effective part of compound Sanhuang tincture was determined. Conclusion:The fingerprints of compound Sanhuang tincture have strong characteris-tics and good reproducibility, which have important reference value for the quality evaluation of ethanol parts of compound Sanhuang tincture.

Objective To observe the microstructural changes in lesions of alopecia areata (AA) with dermoscopy and to evaluate their correlation with clinicopathological manifestations. Methods The area of alopecia of 62 patients with AA and 44 patients with other types of hair loss were observed by using a noncontact polarized dermoscope (Dermlite, USA). Clinical data on and laboratory findings from these patients were collected. Pathological examination was carried out with scalp biopsy specimens from the alopecia area of 15 AA patients. Results Characteristic dermoscopic signs of AA included yellow dots, black dots, broken hairs, exclamation mark hairs, short vellus hair and newly-grown short hairs. Among these signs, yellow dots showed the highest prevalence (83.9%). Exclamation mark hairs, black dots and broken hairs were rather specific signs for AA, and the prevalence of the three signs was positively correlated with disease activity and positivity rate of hair-pull test. A positive correlation was also noted between the prevalence of elevated thyroid peroxidase antibody levels and positivity rate of hair-pull test (r = 0.269, P ＜ 0.05 ) as well as prevalence of broken hairs (r = 0.445, P ＜ 0.05), and between the prevalence of yellow dots and that of keratinous plug in follicular orifice. There was a negative correlation between the prevalence of newly-grown short hairs and perifollicular mast cell infiltration and between the prevalence of black dots and the anagen/catagen ratio. Conclusions Yellow dots can serve as a preliminary screening marker for AA. Exclamation mark hairs, black dots and broken hairs are highly sensitive for the confirmation of diagnosis of AA, and often predict progressive AA.Dermoscopic signs are well correlated to the histopathology features of AA, and may be useful for the evaluation of disease severity and guidance on the treatment of AA.

Objective Establishing a fingerprint method to identify the characteristic chemicals in the roots of Gentiana macrophylla and evaluate their quality.Methods RP-HPLC was developed for fingerprint analysis and determination of four ingredients in G macrophylla roots from different sources.LC-ESI-TOF-MS was employed to identify the chromatographic peaks of the fingerprint.Results Five common peaks were identified by comparing their retention time with reference secoiridoid glucosides.Eight major peaks in chromatographic fingerprint were analyzed by on-line LC-ESI-TOF-MS.Four secoiridoid glucosides were identified based on their MS data.Conclusion The method is specific and could be served for the quality identification and comprehensive evaluation of G macrophylla.

Objective To observe the effect of nerve growth factor(NGF) on CCL4‐induced hepatic fibrosis in mice .Methods The hepatic fibrosis model was induced by subcutaneous injection of CCL 4 in mice .Thirty female Kunming mice were equally and randomly divided into three groups :fibrosis model group (A) ,NGF intervention group (B) and normal saline control group (C) .At 8 weeks following the initiation of experiment ,the samples were collected to measure ALT ,AST ,TBIL ,ALB by the fully automativ biochemical analyzer ,an the liver fibrosis indices (HA ,LN ,PC Ⅲ ) by radioimmunoassay .The Ishaki scoring system was adopted to assess the severity of hepatic inflammation and fibrosis degree .Results Serum levels of ALT ,AST ,HA and LN in the group A and B were significantly higher than those in the group C (F= 111 .45 ,658 .80 ,157 .43 ,167 .99 ;P< 0 .05) ,the levels of ALT 、AST and LN in the group B were significantly lower than those in the group A (P< 0 .05) .The HE staining ,reticular fiber staining and Masson staining showed that the liver fibrosis degree and the liver tissue inflammation in the group A were most obvious ,the liver tissue inflamation in the group B were significantly alleviated as compared with the group A .No fibrous septum was formed and the fiber tissues were fine and short .No obvious inflammatory cells infiltration and fibers formation were found in the liver tissue of the group C .The scores of liver inflamation grade and fibrosis staging in the group C were higher than those in the group B and C ,moreover the scores of liver inflammation grade and fibsosis had statstical differences among 3 groups (P < 0 .05) .Conclusion NGF can block hepatic fibrosis induced by CCL4 and relieve the liver inflammation .

0.05).Microscopic morphology has also been observed, no visible damage could be found in the structure of lenses from eyes injected with ouabain (figs 3 and 4). Serial sections of paraffin-embedded lenses show that the number of fiber cells increased significantly in experimental samples treated with ouabain at a concentration of 0.1 ?M(table 4, P

Objective:To explore the relationship between metabolic acidosis and cardiac valve calcification in maintenance hemodialysis (MHD) patients in the Pearl River Delta Region.Methods:Patients on MHD greater than 3 months who were treated in 10 blood purification centers in the Pearl River Delta Region from July 1 to September 30, 2019 were selected for this multicenter cross-sectional study. Based on a Doppler ultrasound, MHD patients were further divided into non-valve calcification group and valve calcification group. The demographics data, frequency of dialysis, blood pressure, single pool Kt/V(spKt/V), dialysis medications and laboratory data were collected and compared. Spearman correlation analysis was used to analyze the correlation between serum carbon dioxide combining power (CO 2CP) and cardiac valve calcification. Multivariate logistic regression model was used to analyze the influencing factors of cardiac valve calcification. Results:A total of 664 MHD patients were included in this study, with age of (57.0±14.2) years old and dialysis age of 43.0 (22.3, 71.7) months, including 395 males (59.5%) and 269 females (40.5%). Among them, there were 119 patients (17.9%) with diabetes and 186 patients (28.0%) with dialysis 2 times per week. There were 329 patients (49.5%) in the valve calcification group, and 335 patients (50.5%) in the non-valve calcification group. Compared to those in non-valve calcification group, valve calcification group had longer duration of dialysis, higher proportion of patients with dialysis 2 times per week, higher levels of diastolic blood pressure, fasting blood glucose, intact parathyroid hormone and ferritin, higher proportion of patients with blood CO 2CP<19 mmol/L (median CO 2CP), higher proportion of patients on usage of calcium channel blocker, angiotensin converting enzyme inhibitor/angiotensin receptor blocker, α-receptor blocker, β-receptor blocker, calcitriol and lanthanum carbonate (all P<0.05), while the levels of spKt/V, hemoglobin, serum CO 2CP, corrected calcium, blood phosphorus, blood alkaline phosphatase, albumin, total cholesterol, triacylglycerol, low-density lipoprotein, high-density lipoprotein, transferrin saturation, and the proportion of patients on usage of sevelamer and cinacalcet were lower (all P<0.05). Spearman analysis showed significant negative correlation between serum CO 2CP and valve calcification ( rs=-0.697, P<0.001). Multivariate logistic regression analysis showed that dialysis performed twice a week ( OR=2.789, 95% CI 1.232-6.305, P=0.014), blood total cholesterol ( OR=1.449, 95% CI 1.014-2.071, P=0.042), CO 2CP<19 mmol/L ( OR=22.412, 95% CI 10.640-47.210, P<0.001) were the influencing factor of valve calcification in MHD patients. Conclusions:MHD patients with cardiac valve calcification have significant acid loading. Metabolic acidosis is an independent influencing factor for cardiac valve calcification in MHD patients.

OBJECTIVEThis study investigated the role and mechanism of calcineurin (CaN)-nuclear factor of activated T cells (NFAT) pathway in the myoblast apoptosis induced by cyclic tensile strain.

METHODSMyoblasts were cultured using an in vitro-mechanical stimulation model and imposed with tension for different hours with a multi-channel cell stress loading system. Cyclosporine (CsA) was used as CaN inhibitor to clarify the role of CaN in the apoptosis induced by cyclic stress. Hochest 33258 staining and flow cytometry detection were performed to detect the apoptotic cells. Real-time polymerase chain reaction was conducted to detect the mRNA expression of CaN and NFAT. Protein levels of NFAT3 were evaluated by Western blot.

RESULTSThe apoptosis rate increased with the extension of loading time. The mRNA expression of the CaN subunits, CnA and CnB, and the protein levels of NFAT3 also increased. When the myoblasts were incubated with CsA, the apoptosis rate decreased, the mRNA expression of CnA and NFAT3 significantly decreased, and the NFAT3 protein expression levels became significantly lower than those of the groups without CsA.

CONCLUSIONContinuous cyclic tensile stress can induce myoblast apoptosis. The CaN-NFAT signaling pathway may be involved in the cyclic stretch-induced apoptosis of myoblasts.

Apoptosis ; Calcineurin ; genetics ; Cyclosporine ; Flow Cytometry ; Myoblasts ; physiology ; NFATC Transcription Factors ; metabolism ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; T-Lymphocytes

BACKGROUND:Endoplasmic reticulum stress participates in the occurrence and development of many diseases, such as atherosclerosis, diabetes, and Alzheimer’s disease. GRP78 is a marker of endoplasmic reticulum stress. The expression of GRP78 reflects the degree of endoplasmic reticulum stress.
OBJECTIVE:To investigate the effect of cyclic stretch on GRP78 expression of L6 rat myoblasts, and to identify the relationship between cyclic stretch and endoplasmic reticulum stress.
METHODS:In vitro culture-tensile stimulation models of myoblasts of L6 rats were established successful y. The expression of GRP78 of myoblasts exposed to cyclic stretch was determined by reverse transcription-PCR and western blot assay. Stretch groups were subjected to 15%surface elongation at a frequency of 10 cycles per minute, over a period of 1, 6, 12 and 24 hours. cells were simultaneously seeded on a plate in the control and experimental groups with no stimulation.
RESULTS AND CONCLUSION:The expression of GRP78 mRNA was continuously elevated over time after stretched treatment, and significant differences were detected as compared with the control group (P<0.05). GRP78 protein expression began to increase at 1 hour after stretched treatment, was significantly increased at 6 hours, peaked at 24 hours, and significant differences were visible as compared with the control group (P<0.05). In conclusion, cyclic stretch induced the occurrence of endoplasmic reticulum stress, which was enhanced with prolonged time. However, prolonged stretch caused severe endoplasmic reticulum stress and leaded to apoptosis of myoblasts.