Objective To research the effects of drought stress on activity of key enzyme in saikosaponin biosynthesis and saponins content in Bupleurum chinense; To reveal response of Bupleurum chinense to drought stress from protein level. Methods Bupleurum chinense was cultivated in potted water control experiment with 70%–80%, 60%–70%, 40%–50% and 20%–30% saturated soil water contents. The enzyme activities of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR), isoprenyl-based coke (IPPI), farnesyl pyrophosphate synthase (FPS) and β-carotenoid synthase (β-AS) of six months and one year old Bupleurum chinense were measured. The contents of saikosaponins a and d in samples of different soil water contents were determined by HPLC. Results When the saturated moisture was 40%–50%, the enzymatic activities of HMGR, IPPI, FPS and β-AS were the highest, followed by saturated moisture 60%–70%、70%–80% and 20%–30%. The trend of saikosaponin was similar to that of the activity of key enzyme. Conclusion 40%–50% soil saturated water content can significantly increase the activity of the key enzymes in the saikosaponin synthesis pathway. The saponin content and enzyme activity show a significant positive correlation, indicating that under drought stress, Bupleurum chinense regulates the synthesis of saikosaponin by regulating the key enzyme activity of saikosaponin synthesis pathway to respond drought stress.

BACKGROUND:The process of oxidative stress that impacts the curative effect exists in the region which accepts cel transplantation. However, there are few reports about the effects of oxidative stress on human dental pulp stem cels and relevant mechanism.
OBJECTIVE:To understand the effect of hydrogen peroxide on the senescence of human dental pulp stem cels.
METHODS:Human dental pulp stem cels were isolated and cultured in PBS, 100 and 200 μmol/L hydrogen peroxide for 2 hours, respectively. Cel morphology was observed under inverted microscope, degree of cel senescence monitored by β-galactosidase staining, cel proliferation ability detected by BrdU kit and cel counting method, cytoskeleton of dental pulp stem cels and expression of sirt1 tested using immunofluorescence method, and expression of sirt1 and p16 proteins measured by western blot assay.
RESULTS AND CONCLUSION:Dental pulp stem cels exhibited a fibroblast-like morphology with spindle-shaped appearance. After stimulated by hydrogen peroxide, the cel volume was enlarged, theβ-galactosidase staining deepened and the proliferation of dental pulp stem cels reduced. The enhancement of senescence of dental pulp stem cels was accompanied with the increasing concentration of hydrogen peroxide, and in this process, the expression of p16 was raised while the expression of sirt1 was decreased. In conclusion, the senescence of human dental pulp stem cels can be promoted by the stimulation of hydrogen peroxide, and sirt1 and p16 are involved in this process. Our findings may provide a theoretical and experimental foundation for autologous transplantation of dental pulp stem cels.

ificantly lower than in both middle and low group(P＜0.05).Conclusion TP may improve the antioxidant ability of experimental aging mice.

OBJECTIVETo study the relationship between Chinese medicine syndrome and serum level of high-sensitivity C-reactive protein (hs-CRP) in patients with stable coronary heart disease (CHD).

METHODSSerum hs-CRP level was determined in 346 patients with stable CHD, whose diagnoses were confirmed by coronary angiography, and the correlation between patients' syndrome and serum hs-CRP level was analyzed.

RESULTSIn all the CHD patients enrolled, the most commonly encountered Chinese medicine complex syndromes were the qi deficiency and blood-stasis syndrome, the yang deficiency and blood-stasis syndrome, and the qi deficiency with blood-stasis and turbid phlegm syndrome; the dominant syndrome types were blood-stasis, qi deficiency and phlegm-turbidity, which revealed in 324 patients (93.6%), 189 patients (54.6%) and 140 patients (40.5%), respectively. Comparisons of hs-CRP level between different complex syndromes and syndrome elements showed no significant difference (P > 0.05). However, for patients with phlegm-turbidity syndrome, the hs-CRP level was much higher in patients tended to heat than in those tended to cold (2.23 +/- 2.12 mg/L vs 1.59 +/- 1.27 mg/L, P < 0.05). Besides, the score of blood-stasis syndrome showed no correlation with hs-CRP level (Person correlation coefficient: 0.069, P = 0.203).

CONCLUSIONThe elevation of hs-CRP level is closely cor related with phlegm-heat syndrome in stable CHD patients. As an inflammatory biomarker, hs-CRP may be regarded as one of the microcosmic indices of toxin in Chinese medicine, which provides an objective evidence for the pathogenesis hinge of accumulated heat to transform toxin, so it is worthy of further study.

Adult ; Aged ; C-Reactive Protein ; metabolism ; Coronary Angiography ; Coronary Disease ; blood ; diagnosis ; Diagnosis, Differential ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged

Objectlve To observe the influence of Tourmaline on peripheral neuropathy with diabetes mellitus(DM).Methods Tourmaline bedding series were applied to pafients with DM peripheral neuropathy,observeA the changes of subjective symptoms and nervous function,and thus analyzed the therapeutic effects of Tourmaline on DM peripheral neuropathy.Results AfarTourmalinewas usedfor halfa year,some symptoms of the diseasc were alleviated,and after one year's usage.symptoms of spontaneous pain,numbness.and burning sensation improved greatly.Body examinations showed tendon reflex.vibration sense,nerve conducfion velocity,and autonomic nerve were improved on different levels,and progress of disease was obviously coming down.Conclusion Tourmaline bedding series has the function of improving peripheral neuropathy,whichis aconvenientand safety healthcaremethodto CalTyon.

·AlM: To investigate pathogeny and effects of surgery on paralytic strabismus.
· METHODS: A retrospective study was done in 46 patients with paralytic strabismus who underwent squint correction in our hospital from June 2010 to June 2013. Among 26 horizontal strabismus, the cases of extra rectus palsy was 16, internal rectus palsy was 10.Among all20 vertical strabismus, the cases of superior oblique palsy, superior rectus palsy, inferior rectus palsy, double elevator palsy counted for 7, 8, 2 and 3, respectively. Pathogenesis: trauma was 19 cases, followed by 10 cases that the causes could not be identified.Nine was congen ital paralytic strabismus, 8 o ccurred after nose or brain surgery. The surgery methods included rectus muscle recession, rectus muscle resection, partial rectus muscle transposition, Jensen procedure, inferior oblique myectomy and anterior transposition of inferior oblique. Statistical software SPSS10.0 was used in chi-square test between two groups, while the situation of paralysis eye movements improved by two methods in the horizontal strabismus group was compared with t test.
· RESULTS: Among all horizontal strabismus the rate of cure, improvement and inefficiency was 20 ( 77%) , 5 ( 19%) and 1 ( 4%) , respectively. Among vertical strabismus the ratio of cure, improvement and inefficiency was 15 (75%), 3 (15%) and 2 (10%).There was no significantly difference between the two groups ( P >0.05 ). The movements of paralytic eyes were improved. Two procedures used in horizontal strabismus, can improve paralysis eye movements were 3.76 ±0.91, 3.72 ±0.84mm, with no significant difference (P=0.93) statistically.
· CONCLUSlON: Paralytic strabismus in adults had complicated conditions. Choosing different operation methods in treating paralytic strabismus according to the degree of paralysis can result in satisfactory cosmetically alignment of the eyes and modify head position and diplopia.

In order to investigate the biological effects of the VP22?-mE6?/mE7 built-in adjuvant fusion protein vaccine on the tumor associated with HPV-16 infection.HSV-1 VP22? and HPV-16 mE6?/mE7 genes were cloned,and the pET28a-VP22?-mE6?/mE7 recombinat prokaryotic expression vector was constructed.Vector was transformed into Rosetta(DE3)E.coli string and expressed under the induction of IPTG.The re-naturalized protein was then purified via Ni2+ affinity adsorbent column and identified by SDS-PAGE and Western blot.Purified protein was immunized BalB/C and C57BL/6 mice to evaluate the immunogenicity and anti-tumor activity.The expressed recombinant protein formed as inclusion body with a prediction MW about 34kDa and contained approximately 45% of total somatic protein.The VP22?-mE6?/mE7 immunization induced higher titer of specific IgG against HPV,higher level of lymphocyte proliferation and better effect on suppressing HPV16 positive TC-1 tumor growth than the mice immunized with mE6?/mE7 alone.The results showed that the recombined built-in adjuvant vaccine could induce specific cellular immune response in vitro and inhibit the TC-1 tumor proliferation in vivo,that would be a foundation for further studies and developments of inner adjuvant vaccines.

OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

OBJECTIVETo observe the effect of Yangjing Zhongyu Decoction (YZD) on mRNA and protein expression of PCNA, StAR, and FSHR in ovarian granulose cells (GCs) cultured by excess androgen.

METHODSOvarian GCs from porcine follicles were isolated and cultured in vitro. Follicular stimulating hormone (FSH) or YZD was added in the GCs treated by excess testosterone propionate. Totally 48 h later mRNA and protein expression of PCNA, StAR, and FSHR were detected by RT-PCR and Western blot.

RESULTSExcess androgen inhibited mRNA and protein expression of PCNA, StAR, and FSHR of GCs. FSH and YZD could antagonize inhibition of excess androgens, and promote mRNA and protein expression of PCNA, StAR, and FSHR in GCs.

CONCLUSIONYZD could antagonize the inhibition of excess androgen on mRNA and protein expression of PCNA, StAR and FSHR in GCs. Thus, we inferred that YZD could improve the follicle dysplasia by promoting mRNA and protein expression of PCNA, StAR and FSHR in GCs.

Androgens ; pharmacology ; Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Membrane Transport Proteins ; genetics ; metabolism ; Ovarian Follicle ; cytology ; drug effects ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics ; Receptors, FSH ; genetics ; metabolism ; Swine

In this study,thermosensitive and repairable molecularly imprinted solid-phase microextraction fibers were synthesized using spiramycin as template molecule,methacrylic acid and N-isopropylacrylamide as functional monomers,ethylene glycol dimethacrylate as crosslinking agent,and silanized quartz capillary as carrier. The prepared molecularly imprinted solid-phase microextraction fibers were characterized by scanning electron microscope and nitrogen adsorption/desorption,and various parameters affecting the extraction efficiency were optimized. Due to high selectivity and sensitivity of the fibers for macrolide antibiotics,the quantitative analysis of four macrolide antibiotics in food matrix,spiramycin,tilmicosin,tylosin,and josamycin,was peroformed in combination with high performance liquid chromatography. In the range of 0.5 to 50 μg/mL,the chromatographic peak area showed a good linear relationship with the concentration. The spike recoveries of the samples at three different addition levels were between 81.8% and 119.1%;the inter-day precisions were less than 13.8% (n=6),and the intra-day precisions were less than 15.5% (n=3).