Objective To develop a method for determining verbascoside in Mitonghua Granules by HPLC. Method Verbascoside was determined by HPLC method,Agilent ZORBAX-Extend C18 column (4.6 mm?150 mm,5?m) with Acetonitrile-1 %Acetic acid(13∶87) as the mobile phase was used for elution.The flow rate was at 1.0 mL/min and the detective wavelength was at 334 nm. The column temperature was at 30 ℃. Results There was a good linearity relationship at the range of 0.062 8～1.256 0 ?g for verbascoside (r=0.999 83) .The average recovery was 99.61 %,and RSD was 1.55 %(n=6). Conclusion The method is simple,reproducible and accurate with a good repeatability,and can be used for the quality control of Mitonghua Capsules.

OBJECTIVETo evaluate the functional outcomes of modified limited "L" incision beside the Achilles tendon with distraction bone block arthrodesis in treatment of subtalar osteoarthritis.

METHODSFrom March 2009 to September 2012, a total of 22 cases of old calcaneus fractures with subtalar osteoarthritis were treated with modified limited "L" incision and distraction bone block arthrodesis including 13 males and 9 females with a mean age of 35.3 years old (ranged 22 to 49). The mean time from calcaneal fracture was 21 months (ranged 11 to 32). According to the Stephens-Sanders classification, 16 cases were type II and 6 were type III. The modified-AOFAS ankle-hindfoot score was used for functional outcomes evaluation.

RESULTSThere was one incision necrosis and no infection, implant failure, bone-graft absorbed or talus necrosis was note at the follow-up time. A total of 21 cases were followed up for a mean time of 29 months (ranged from 18 to 46 months). All of the cases reached a bony union within 4 months postoperation. The mean modified-AOFAS ankle-hindfoot score was 82.6 points (ranged from 66 to 92 points),reached a significantly improvement in comparing with the mean preoperative score (50.8 points,ranged from 32 to 65 points, P < 0.01).

CONCLUSIONThe modified limited"L" incision beside the Achilles tendon with distraction bone block arthrodesis is an acceptable and alternative treatment method for subtalar osteoarthritis. This method is easy to use and with less complication. It can correct the main pathological changes and reach good functional outcomes.

Adult ; Arthrodesis ; methods ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis ; surgery ; Subtalar Joint ; surgery

BackgroundGlaucoma is an optic neuropathy caused by structural damage of the optic nerve,and its early diagnosis is critical for arresting the irreversible damage of visual function. Optical coherence tomography (OCT) allows an early diagnosis of glaucoma by the measurements of the optic disc and retinal nerve fiber parameters. Objective This study was carried out to evaluate the effects of optic disc tomography and the measurement of the retinal nerve fiber layer (RNFL)thickness by spectral-domain OCT on the diagnosis of glaucomatous eye. MethodsIt was a noninterventional, cross-sectionalstudy. The optic disctopographic parameters and total and regional RNFL thickness were measured by RTVue OCT in 62 normal eyes and 67 glaucomatous eyes. The area under the receiver operating characteristic curve( ROC ) was used to assess the ability to differentiate glaucoma eyes from normal eyes of each testing parameter. This trial complied with the Helsinki Declaration and was approved by the Clinical Trial Ethic Committee of Beijing TongrenHospital. All of the participants signed the written informed consent before any medical examination. Results In the comparison of demography ,the ages of patients, the mean deficiency( MD ) and pattern standard difference( PSD ) of perimetry were obviously larger in the glaucoma group, primary open angle glaucoma ( POAG ) group and primary closure-angle glaucoma(PACG) group than those of normal controls( P＜0. 01 ). No significant differences were found in the disc area between a total glaucoma group, POAG group or PACG group and normal group ( P =0. 101,0. 741 and 0. 652, respectively) ;however, the average RNFL thickness between normal eyes and glaucomatous eyes were significantly different( 109. 758 μm versus 79. 539 μm, P＜0. 01 ). Among the eight regions around the optic disc, the thickest RNFL located at the inferotemporal( 150. 109 μm) and superotemporal( 146. 105 μm) regions in normal eyes,and at the superotemporal( 104. 354 μm) and inferotemporal( 102. 436 μm) regions in glaucomatous eyes. Both in normal and glaucomatous eyes,the thinnest RNFL located at the nasal(NU+NL) and temporal(TU + TL) regions. For optic disc topographic parameters,the highest ROC were observed in rim volume( ROC--0. 850,0. 841 and 0. 862 in total glaucoma,POAG and PACG, respectively) and vertical cup/disc ratio( ROC =0. 840,0. 849 and 0. 830 in total glaucoma,POAG and PACG,respectively), and the sensitivities for specificity cutoff set at 80％ were 73.1％ and 76. 1％ in total glaucoma,73.0％ and 81.1％ in POAG and 73.3％ and 70.0％ in PACG, respectively. For RNFL thickness ,the highest ROC was observed in average RNFL( ROC =0. 925,0. 910 and 0. 942 in total glaucoma, POAG and PACG,respectively) ,and the sensitivities for specificity cutoff set at 80％ were 89. 6％ ,89.2％ and 90. 0％ in total glaucoma,POAG and PACG, respectively. Among the eight regions around the optic disc, RNFL thickness of region IT achieved the highest ROC, RNFL thickness of region TU and TL had the lowest ROC. Conclusions RTVue OCT appears to be of fair discriminating ability in distinguishing normal from glaucomatous eyes. RTVue OCT shows promise for the diagnosis of glaucoma.

AIM: To analyze and identify the phosphoproteins associated with diazoxide preconditioning. METHODS: Proteomics technique was used to investigate the changes of phosphoprotein after diazoxide preconditioning. Adult rat ventricular myocytes were pretreated in the presence and absence of 200 ?mol/L diazoxide for 10 min. Phosphoproteins prepared and enriched respectively from control and diazoxide pretreated groups were then separated by two-dimensional (2D) gel electrophoresis and stained with sliver staining kit. Phosphoproteins of interest were further identified by mass spectrometry. RESULTS: Associated with diazoxide preconditioning, the proteins of chaperonin containing TCP-1 and hypothetical protein XP_346548 were phosphorylated significantly. The proteins of 94 kD glucose-regulated protein, calpactin I heavy chain and ferritin were dephosphorylated markedly (P

Objective To study the effect of caffeic acid phenethyl ester (CAPE) on the expression of ?-catenin in the cultured colorectal cancer cell lines. Methods HCT116 and W480 cells were treated with CAPE at serial concentrations of 2.5, 5, and 10 mg/L. ?-catenin protein expression was assayed by Western blot analysis. ?-catenin localization was detected by indirect immunofluorescence. Results CAPE treatment was associated with decreased total ?-catenin protein expression. The expression of ?-catenin at the cell nucleus and cytoplasm was downregulated, but at the cell-cell linked site the ?-catenin protein expression was upregulated. Conclusion CAPE can downregulate the expression of ?-catenin and inhibit the translocation of ?-catenin to nucleus, which may play an important role in the anticancer activity of CAPE.

Objective To explore the nutritional status and determinants among rural stranded children under 7 years of age. Methods The group of stranded children (n=7585) and the children for control (n=7557) were identified by multi-stage stratified cluster sampling. The ascertainment methods mainly included questionnaire, anthropometric measurements and laboratory examination. Results In this study, there were three groups, including the group of stranded children whose single parent left home to look for better job somewhere, the group of stranded children whose parent both left home and the third group serving as control. Prevalent rates on the following items were: stunting -- 14.9%, 17.9% and 16.3%, underweight -- 7.2%, 8.3% and 7.6%, wasting -- 3.1%, 3.4% and 3.3% and anemia -- 18.9%, 20.6%, 18.7% respectively. The prevalent rate of stunting in the group of children with both parents away from home was significantly higher than that those with only one parent did. The prevalence rate of anemia in the group with both parents away from home was significantly higher than that in the control group. Based on multivariate non-conditional logistic regression analysis, the determinants of stunting among the stranded children were: age, with low birth weight, living alone, being the only child in the family, mother's education level, frequency of eating breakfast and snacks every week, motives of caretakers and the relationship between children and the caretakers. The determinants of underweight among those stranded children were: age of child, with low birth weight, the frequency of eating snacks every week, motives of the caretakers and having rickets. Conclusion The nutritional status among rural stranded children was relatively poor, especially in those whose parents were both away from home, which calls for urgent improvement.

Recently, herbal medicine including traditional Chinese medicine (TCM) has gained huge attention in the world. In 2015, the global trades of herbal medicine reached 93.15 billion US dollars. And, the latest statistics from the Ministry of Industry and Information Technology of People's Republic of China showed that total sales of Chinese patent medicine and raw herbs reached 120 billion US dollars in 2014. Therefore, the aim of this study was to analyze the situation of international marketing on herbal medicine and how much TCM shared in it. The PubMed database, search engines and government websites and research reports were searched for analyses. The results showed that total trades of TCM products in both domestic and foreign markets, were about 135 billion US dollars, including Chinese patent medicine, raw herbs, herbal extracts, herbal health care products, whose proportion of the global marketing was 80%.

OBJECTIVETo optimize cultivation methods of bone marrow mesenchymal stem cells (MSCs) from hepatitis B patients and to investigate their biological characteristics.

METHODSGrowth curves of hepatitis B patients MSCs cultivated with five culture media and two inoculation methods were compared; the shapes, appearances, surface markers and bionomics of the cultivated MSCs were studied.

RESULTSInoculating the cells obtained directly from bone marrow aspirations was not as successful as using the marrow cells after their density gradient centrifugations (76% vs 88%), but the differences in the results were not statistically significant (P more than 0.05). The successful cultivation rates using five culture media were different and the differences were statistically significant (P less than 0.01). The autoserum medium was most successful, fatal bovine serum (FBS) medium was next successful and the non-patient serum medium was the least successful. The growth curves of the cultivations using the different media were parallel to this. Changing the whole culture media every 2 or 3 days was better than changing half of the media. The shapes, appearances, surface markers and the growth characteristics of MSCs from the hepatitis B patients were almost the same as MSCs from the normal adult.

CONCLUSIONThe best cultivation method of MSCs from hepatitis B patients is: separating marrow cells using density gradient centrifugal separation, cultivating them using an autoserum culture medium, and completely changing the medium every 2-3 days. The biological characteristics of MSCs from the hepatitis B patients using the above methods are almost the same as those from normal adults.

Adult ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Hepatitis B ; Humans ; Mesenchymal Stromal Cells ; cytology ; Middle Aged

OBJECTIVETo investigate the effect of diet-induced obesity on the apoptosis of testicular spermatogenic cells in pubertal male rats.

METHODSForty healthy male rats were equally and randomly divided into a control and a high-fat group, the former fed on normal diet, while the latter high-fat and high-calorie diet. The testes of the rats were harvested at the end of 10 weeks for detection of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) in the peripheral blood with the automatic biochemical analyzer. Pathological changes of the testis were observed under the light microscope, the apoptosis of the testicular cells detected by TUNEL, the expressions of Bcl-2 and Bax proteins determined by immunohistochemistry, and those of Bcl-2 mRNA and Bax mRNA measured by RT-PCR.

RESULTSThe levels of TC, TG, LDL-C and HDL-C were significantly higher in the high-fat group (5.17 +/- 0.17, 1.18 +/- 0.09, 1.76 +/- 0.11 and 5.08 +/- 0.18) than in the control (1.38 +/- 0.12, 0.39 +/- 0.05, 0.97 +/- 0.07 and 0.75 +/- 0.06) (P < 0.05), so was the apoptotic index of spermatogenic cells (37.17 +/- 2.74 versus 5.16 +/- 0.81, P < 0.01), and the apoptotic spermatogenic cells were mainly spermatogonia and spermatocytes. The expressions of Bax protein and Bax mRNA were markedly higher in the high-fat group (153.26 +/- 8.74 and 1.08 +/- 0.12) than in the control (101.81 +/- 6.14 and 0.37 +/- 0.04) (P < 0.01), while those of Bcl-2 protein and Bcl-2 mRNA remarkably lower in the former (139.26 +/- 7.21 and 0.46 +/- 0.05) than in the latter (159.37 +/- 8.96 and 1.05 +/- 0.11) (P < 0.01).

CONCLUSIONDiet-induced obesity can increase the apoptosis of spermatogenic cells in the rat testis, which may be associated with the reduced expression of Bcl-2 and elevated expression of Bax.

Animals ; Apoptosis ; Diet ; Lipids ; blood ; Male ; Obesity ; etiology ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Testis ; cytology ; pathology ; bcl-2-Associated X Protein ; metabolism

Objective: To investigate the possibility of adipose tissue mesenchymal stem cell-derived microvesicles (ADSC-MVs) to improve the retention volume of fat transplantation. Methods: Human adipose tissue was obtained from 5 healthy female patients aged from 20 to 30 years, who came to the hospital for abdominal liposuction. Adipose tissue mesenchymal stem cells (ADSCs) were acquired by collagenase enzymatic hydrolysis. ADSC-MVs were isolated from the supernatant of cultured ADSCs through ultra-centrifugation, and characterized by transmission electron microscope and PKH26 staining. Sixteen BALB/c-nu nude mice were randomly divided into 2 groups (n=8, mice/group) by random number table method. In EV group, the mice were subcutaneously injected 0.35 ml human fat, with 7 μg ADSC-EVs (final concentration is 20 μg/ml), while 0.35 ml human fat were in blank group. The mice were sacrificed and the grafts were harvested at 1 month and 3 months after transplantation. Weight and volume of transplanted fat were tested. Morphology of adipocytes, fibrosis and necrosis of fat tissue were confirmed by HE staining. Blood vessel density were accessed by CD31 staining.SPSS 13.0 was used for statistical analysis and Student-t test was applied to compare the data from two groups. Results: One month after fat transplantation, the volume of fat grafts in blank group was lower than that in MV group. Besides, lipoliquefaction can be observed in fat grafts of blank group. Fat grafts in MV group had a higher volume and littler necrosis and lipolique faction. Three months after grafting, the volume of fat grafts in blank group [(0.057±0.009) ml] was significantly lower than that in MV group [(0.103±0.015) ml], t=8.117, P<0.001. MV group had an increased number of vessels (29.6±4.0/field) compared with grafts from the Blank group (23.0±2.4/field), t=2.825, P=0.022. Histological analysis revealed that the fat grafts in MV group consisted less fat necrosis and fibrosis than control group. Conclusions ADSC-MVs can improve the retention volume of fat transplantation.