0.05.[Conclusion]VDR TruⅠ and FokⅠpolymorphisms are not related to LDD in Han nationality.

OBJECTIVE: To determine the changes of the plasma metabolic phenotype in rats with chronic restraint stress (rats with syndrome of liver qi stagnation and spleen deficiency), so as to reveal the biological features of the syndrome of liver qi stagnation and spleen deficiency, and to examine potential application of metabonomic analysis in studies of syndromes in traditional Chinese medicine. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into four groups: group A, 7 d normal control group; group B, 21 d normal control group; group C, 7 d stress group; and group D, 21 d stress group, with 6 rats in each group. Chronic restraint was used to induce stress in rats. Blood was collected from the cardio-ventricle under anesthesia on the 8th (groups A and C) or 22nd day (groups B and D) and detected by using the Fourier variable superconducting nuclear magnetic resonance (NMR) spectrometer (Varian UnityInova 600 M). Free induction decay signals were transferred into one-dimensional NMR spectrogram via 32 k Fourier transformation. Segmental integral calculus (0.04 ppm per segment) was performed from 4.5-0.5 ppm (Carr-Purcell-Meiboom-Gill, CPMG) or 6.0-0 ppm (longitudinal eddy-delay, LED) as defaulted 1H spectra values by using the VNMR software. Data were saved as text or excel files after normalization and then used for pattern recognition analyses. All the data were analyzed by principal component analysis (PCA) using the SIMCA-P 10.0 software (Umetrics AB, Umea, Sweden). RESULTS: The PCA analysis of rat plasma (1)H NMR spectra revealed different metabolic spectra between stress and control groups, which were consistent with alterations of in vivo metabolisms in rats under stress stimuli. Compared with the normal control group, rats with repeated stress displayed significant changes in spectral peak shapes of acetate, lactate, tyrosine, low-density lipoprotein, and unknown compounds (3.44 ppm). These altered metabolites can be used as biomarkers of syndrome of liver qi stagnation and spleen deficiency for further studies. CONCLUSION: The (1)H NMR spectra of metabolites in the rat blood are differentially changed after chronic stress. Specific, characteristic metabolic products can be identified by analyses of metabonomics, which lead to interpretation of biological feature of Chinese medicine syndromes. Therefore, metabonomic analysis is an approach with good development prospects to studies of TCM syndromes.

A PCR method was used to amplify the sequence encoding the mature peptide of?-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream of?-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU/ml. The properties of the recombinant mannanase were characterized.

Objective To evaluate clinical efficacy of multiple regimen combination in treatment of osteoporosis of perimenopausal or postmenopausal women. Methods From Jul. 2008 to Dec. 2009, 109 women with low bone mineral density (BMD) or osteoporosis treated in Department of Obstetrics and Gynecology, Affiliated Second Hospital, Wenzhou Medical College were enrolled randomly into 3 group,including 36 women in Group A managed by osteoform 1000 mg/d + alfacalcidol 0. 25 μg/bid orally, 40 women in group B managed by osteoform 1000 mg/d + alfacalcidol 0. 25 μg/bid + tibolone 1.25 mg/d orally and 33 women in group C managed by ostcoform 1000 mg/d + alfacalcidol 0. 25 μg/bid +bisphosphonates 70 mg/w orally. After 48 weeks BMD on lumbar 1 -4 (L1-4) and left femur were detected by X-ray. Bone alkaline phosphatase(BALP) ,cross linked clelopeptide of type Ⅰ collagen(CTX) and 25-hydroxychole calciferol [25 (OH) D3] was measured by enzyme linked immunosorbent assay (ELISA).Result Seven women (6. 4%, 7/109) were withdrawed form this study, including 2 cases losing follow up in group A, 3 cases stopping treatment in group B, 2 cases giving up treatment due to severe adverse effect (burning in upper abdomen) in group C. (1) Pain relieve: after 48 weeks treatment, women in 3 groups improved symptom of pain significantly, the rates of pain relieve were 85% (29/34)in group A, 92% (34/37) in group B and 94% (29/31) in group C. (2) BMD: BMD was improved significantly in women in 3 groups after treatment. BMD of L1-4 were (0.88±0.15) g/cm2 in group A,(0.89±0.18) g/cm2 in group B and (0.87±0.10) g/cm2 in group C before treatment, and converted to (0.90±0.01) g/cm2 in group A, (0.93±0.09) g/cm2 in group B and (0.91±0.11) g/cm2 in group C after treatment. BMD of left femur were (0.87±0.07) g/cm2 in group A, (0.87±0.07) g/cm2 in group B and (0.85±0. 12) g/cm2 in group C before treatment and converted to (0.90± 0.03) g/cm2 in group A, (0.91±0.08) g/cm2 in group B and (0.89 ±0.12) g/cm2 in group C after treatment. It was shown significantly different BMD between group B or C and group A (P < 0. 01), however, there was no significant different BMD between group B and C (P >0. 05). (3) Index of bone metabolism: BALP were (26±6) μg/L in group A, (26±9) μg/L in group B and (28±7) μg/L in group C before treatment and converted to (22±5) μg/L in group A, (20±9)μg/L in group B and (22±8)μg/L in group C after treatment, which showed statistical difference (P < 0.05). CTX were (0.85±0.20) ng/L in group A, (0.84±0.47) ng/L in group B, and (0. 88 ±0. 11) ng/L in group C before treatment and converted to (0. 81 ±0. 19) ng/L in group A, (0. 77±0.33) ng/L in group B, and (0.82 ±0. 14) ng/L in group C after treatment, which showed statistical difference (P < 0. 05). Conclusions Those 3 regimens combination could be used in treatment of osteoporosis by decreasing bone conversion, increasing bone density, decreasing bone absorption. Regimen A was only suitable for basic therapy,the other two regimens could provide better treatment.

Objective To investigate the expression and significance of JWA in Acute Myeloid Leukemia (AML). Methods Bone marrow mononuclear cell specimens were taken from 22 AML patients in newly diagnosis stage and complete remission stage respectively , and the JWA expression were detected at RNA and protein level. Results (1) JWA was expressed in all the samples at RNA and protein level. (2) At protein level, JWA expression was higher in the cells from newly diagnosed AML patients than in those from patients in complete remission stage (P < 0.05). (3)The complete remission rate of patients with higher expression level of JWA (87.5%) was similar to those with lower expression (83.3%). The complete remission rate of patients with higher expression level of JWA after the first course (31.25%) was lower than those with lower expression (66.7%). The early recurrence rate of patients with higher expression level of JWA (42.86%) was higher than those with lower expression (20%). Conclusion JWA may play an important role in the development and progression of AML. Increased expression of JWA may be one of the causes of refractory and relapsed AML.

Objective To explore the role of brief ischemia pretreatment in the induction of renal ischemic tolerance,and investigate its effects on tubular cell necrosis,apoptosis and proliferation. Methods Male Sprague-Dawley rats were randomly divided into three groups,including sham-operated group (Sham),ischemia/reperfusion injured group subjected to theocclusion of both renal pedicles for 40 min followed by reperfusion(I/R),and preconditioned group with 20-min ischemia pretreatment induced 4 days before I/R(IPC).Histological changes were evaluated by PAS staining.The ultra-structure of tubular cells was observed by transmission electron microscopy(TEM).Apoptosis was confirmed by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL).The proliferation of tubular cells was evaluated with proliferating cell nuclear antigen(PCNA). Results Twenty-minites ischemia pretreatment offered both promising functional and histological protection against 40-min ischemia/reperfusion injury (P<0.01).The mortality rate wag reduced from 33%in I/R group to 0 in IPC group.The renopmtection offered by 20-min ischemia pretreatment was accompanied with reduced postischemic tubular cell apoptosis and necrosis (P<0.05), and increased cell proliferation (PCNA positive) (P< 0.01). Conclusions Brief and sublethal prior ischemia can render the kidney more tolerant to subsequent prolonged I/R injury. Its ability to tilt the balance of tubular cell fate toward survival, reducing postischemic cell death and enhancing cell proliferation, may play an important role in renal protection of ischemic preconditioning.

Background and purpose: Increasing evidence has indicated that polymorphisms in the microRNA (miRNA, miR) binding site of target gene can alter the ability of miRNA and modulate the risk of cancer. We aimed to investigate the association between a miR-520a binding site single nucleotide polymorphism (SNP) rs141178472 in the PIK3CA 3’UTR and the risk of colorectal cancer in a Chinese Han population. Methods:The polymorphism rs141178472 was analyzed in a case-control study, including 386 colorectal cancer patients and 394 age-and sex-matched controls. The relationship between the polymorphism and the risk of colorectal cancer was examined by statistical methods. Results:Individuals carrying the rs141178472 CC genotype or C allele had an increased risk of developing colorectal cancer (CC vs TT, OR=1.716, 95%CI:1.084-2.716, P=0.022;C vs T, OR=1.258, 95%CI:1.021-1.551, P=0.033). Furthermore, the expression of PIK3CA was detected in the peripheral blood mononucleated cell of colorectal cancer patients, suggesting that mRNA levels of PIK3CA might be associated with SNP rs141178472. Conclusion:These ifndings provide evidence that a miR-520a binding site polymorphism rs141178472 in the PIK3CA 3’UTR may play crucial roles in the etiology of colorectal cancer.

AIM: To observe effect of the iris location to femtosecond - combined wavefront guided LASIK for myopia and astigmatism.?METHODS:The patients with astigmatism >1. 0D during the same time and followed up for 1a were selected. A total of 129 eyes in 67 patients were treated under iris location with femtosecond-combined wavefront guided LASIK ( experimental group) and 161 eyes in 82 cases with femtosecond-combined wavefront guided LASIK ( control group) . Laser cutting went with the same laser machine. The uncorrected visual acuity ( UCVA ) , best corrected visual acuity ( BCVA) , and wavefront aberration between the two groups were compared at 1, 3, 6mo and 1a after surgery.?RESULTS:At 1 and 3mo after surgery, the number of patients with better postoperative UCVA than preoperative BCVA between the two group showed a statistically significant difference (χ2=6. 423, P=0. 011,χ2=14. 431, P=0. 01 ); at 1d and 1mo after surgery, the residual astigmatism showed a statistically significant difference between two groups (t=1. 98, P<0. 05; t=2. 23, P<0. 05). At 3, 6mo and 1a after surgery, the differences on the change of residual astigmatism between the two groups weren’t significant ( P > 0. 05 ). At 6mo and 1a after surgery, the differences on UCVA between the two groups weren’t significant ( P > 0. 05 ). Until 1a after surgery, the root mean square ( RMS ) of high order wavefront aberration of the two groups, spherical aberration and coma aberration ( COMA ) were all enhanced compared to before surgery(P<0. 05). At 1, 3mo after surgery, the RMS showed a statistically significant difference between two groups (P<0. 05). At 1, 3, 6mo, 1a after surgery, the increase of COMA in experimental group was significantly lower than that in control group (P<0. 05).? CONCLUSION: Iris location technology applied in femtosecond - combined wavefront guided LASIK for myopia and astigmatism, can make the vision recovery faster, the RMS of high order and COMA increase less, the residual astigmatism less, show better and more stable treatment effect.

AIM:To contrast analysis of postoperative variation of corneal posterior surface heights after Femtosecond LASlK ( FS - LASlK ) and small incision lenticule extraction ( SMlLE) for high myopia.
●METHODS: Sixty-seven cases of high myopic patients (132 eyes) operated with laser corneal refractive in our hospital from May to Dec. in 2015 (-6. 00D≤spherical equivalent degree≤-10. 00D) were selected and divided into FS-LASlK group and SMlLE group. The thickness of corneal flaps at FS-LASlK and the thickness of map at SMlLE were designed to be 110μm. Corneal posterior surface heights were examined by Pentacam at preoperation, postoperative 3 and 6mo after FS-LASlK and SMlLE operation. Surface height changes after preoperative, postoperative 3 and 6mo were compared by measuring Pentacam corneal analysis system.
●RESULTS: Six months after operation, the FS-LASlK posterior corneal surface height was 6. 47 ± 1. 65mm, significantly higher than 5. 20 ± 1. 32mm before operation. SMlLE posterior corneal surface height was 6. 40 ± 1. 33mm, significantly higher than 5. 18 ± 1. 25mm before operation, the differences were statistically significant( P<0. 05). Six month after surgery, two methods of corneal surface height variation obtained was 1. 29 ± 1. 28mm and 1. 22 ± 0. 89mm, there was no significant difference ( P>0. 05).
●CONCLUSION:After FS-LASlK and SMlLE, the corneal posterior surface is protrusive. FS - LASlK is slightly obvious than SMlLE in early period. The stability of the posterior surface is better after SMlLE.

Objective To explore the enhancement pattern and characterization of hepatic hemangionmas with contrast enhanced ultrasound (CEUS).Methods A total of 44 patients with 49 nodules preliminary diagnosed of liver hemangioma were included in this study.For each nodule,the enhancement pattern,level,and dynamic change of CEUS were evaluated,and the features of hemangionmas were groups as echoic and compared with those on CEUS.Results All hemangiomas enhanced in arterial phase with centripetal progression in venous and late phase on CEUS,among which 41 lesions showed peripheral nodular enhancement while 8 showed peripheral rim-like enhancement.The whole-tumor enhancement pattern was seen in 13 lesions and part-tumor enhancement was shown in 36 lesions.The performance of part-tumor was independent of tumor echoic and hypoechoic tumors mostly presented to be whole tumor enhancement pattern.During portal venous and late phase,42 lesions showed hyperenhancement,while 7 lesions showed isoenhancement.Conclusions CEUS can suggest the enhancement dynamic characters of hepatic giant hemangionmas and reveals the relationship of grey-scale echoic and enhancement pattern of hemangiomas.It is important to diagnose the hepatic giant hemangionmas for CEUS.