The broad-spectrum antibacterial action of berberine hydrochloride mainly contributes to recurrent aphtha, periapical periodontitis, radioactive mucositis and pericoronitis, however a little evidence support the action mechanism underlying periodontitis treatment. OBJECTIVE: To determine the effects of berberine hydrochloride on the expressions of related cytokines in periodontal tissues of experimental periodontitis rats, to reveal and understand the action pathway of berberine hydrochloride on oral tissue repair. METHODS: Sixty Wistar rats weighing 160-200 g, aged 3 months Models, were involved in this study.Models of experimental periodontitis were established in rats through a use of local steel-wire ligation and systemic injection of prednisone acetate. Forty successfully established models were randomized into periodontitis model group (n=8) and periodontitis treatment group (n=32), at the same time, 10 normal rats served as control group. The treatment group of animals were fed with 0.06 g/kg berberine hydrochloride daily and medicated to death over the 1, 2, 3, 4 weekends (8 rats each). The model group was fed with isodose normal saline. The model group and normal control group were killed at the fourth weekend. Main observations: ①Oral gross observation and X-ray film examination; ②Pathological assay of periodontal tissues; ③Immunohistochemical SABC method was conducted to determine the expressions of tumor necrosis factor-α (TNF-α), bone gla protein (BGP), interleukin-1β (IL-1β), interleukin-6 (IL-6) in periodontal tissues in rats. RESULTS AND CONCLUSION:①Following hormone injection, gum tissue exhibited erosion and pyorrhea in model group of rats; the above-mentioned symptoms were relieved in rats of treatment group; there was no abnormality in periodontal tissues of normal rats. X-ray examination revealed alveolar crest resorption and obvious interredicular shadow in the model group.②Rats of model group showed obvious pathologic changes in periodontal tissues, the levels of TNF-α, IL-1β, IL-6 were significantly higher and the level of BGP was dramatically lower than those in normal group (P < 0.05); Treatment with berberine hydrochloride decreased the levels of TNF-α, IL-1β, IL-6 in periodontal tissues and increased the level of BGP compared with model group (P < 0.05). The periodontal tissues in groups treated with berberine hydrochloride exhibited pathological changes at inflammatory repair stage. Results showed that berberine hydrochloride inhibits the expressions of TNF-α, IL-1β, IL-6 in periodontal tissues in experiment rat models of periodontitis, and promotes the expression of BGP and repair of periodontal tissue.

OBJECTIVETo evaluate the effects of Icariin (ICA) on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in vitro and in vivo.

METHODSAn enzymatic digestion block was used in vitro to culture hPDLSCs, which were separated and purified by limited dilution cloning. The hPDLSCs were identified using cell-surface markers and cocultured with 1 x 10(-7) mol.L-1 ICA solution. The proliferation ability of these cells was determined by thiazolyl blue tetrazolium bromide (MTT) assay. After staining with alkaline phosphatase (ALP), osteogenesis was detected by enzyme-linked immunosorbent assay. Osteoblast-related genes were analyzed by reverse transcription-polymerase chain reaction. Alizarin red staining was performed to measure the level of calcium deposition. The hPDLSCs were cocultured with 1 x 10(-7) mol.L-1 ICA and nano-hydroxyapatite scaffolds in vivo before transplantation into subcutaneous tissues of nude mice. Osteogenic abilities were histochemically analyzed after 30 days of induction.

RESULTSThe hPDLSCs were affected by 1 x 10(-7) mol.L-1 ICA, and MTT assay showed that the proliferation of the groups treated with ICA in vitro was better than that of the control groups on the second day. The ALP activity of the treated hPDLSCs was significantly enhanced after cell culture for 3, 5, and 7 days. The gene expression of osteoblastic markers was also significantly enhanced after 7 days. The deposition of mineralization after incubation with 1 x 10(-7) mol.L-1 ICA increased compared with the control after cell culture for 14, 21, and 28 days. Furthermore, the bone expression of the treatment groups in vivo was significantly enhanced compared with that of the control groups.

CONCLUSIONTreatment with 1 x 10(-7) mol.L-1 ICA can significantly promote proliferation and differentiation of hPDLSCs in vitro and in vivo. ICA can effectively function as a bioactive growth factor in periodontal tissue engineering to replace traditional growth factors.

Alkaline Phosphatase ; Animals ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Coculture Techniques ; Enzyme-Linked Immunosorbent Assay ; Flavonoids ; Humans ; Mice ; Mice, Nude ; Osteoblasts ; Osteogenesis ; Periodontal Ligament ; Stem Cells

Objective To exlore the influence of internal quality control and external quality control assessment(EQA) resulting from applicability of control samples in measurement of whole blood viscosity (WBV) through the analysis and comparison of applicability of 1 non-Newtonian fluid internal quality control sample in 3 viscometers. Methods Viscometer B, C and D were used to measure WBV of 30 blood samples in parallel under the shear rate(SR) of 1 s-1,30 s~(-1) and 200 s~(-1), then the blood SR-WBV curves of 3 viscometers were drawn according to the results. At the same time, viscometers B, C and D were used respectively to determine the WBV of control A 10 times in one day, then the control A SR-WBV curves were mapped. Three viscometers were used to measure the manufactory control samples and control A 5 times in one day for 4 days. Four groups of daily values of manufactory control samples and control A of each instrument were used to carry out F test to calculate whether 4 daily values are difference. Finally, the control A was dispensed in 49 laboratories nationwide chosen for measurement. On the basis of viscometer used, 20 laboratories were classified as group B, 20 laboratories were classified as group C and 9 laboratories were classified as group D. Then the data under SR of 1 s~(-1) were analyzed to calculate the coefficient of variation (CV) in the group. Results There was significant difference among the WBV of blood samples measured by the viscometers B, C and D. The results under SR of 1 s~(-1) declined in turn, and they were highest under SR of 30 s~(-1) followed by the values of viscometer D and B and they were (8.14±0.75), highest under SR of 30 s-1 followed by the values of viscometer B and D, and they were (7.35±0.07), daily values of manufactory control and control A of each instruments in four groups were compared. Under SR of 1 s~(-1), there was no difference between daily values of manufactory control and control A in viscometer B (F = 2.63, 1.37, P > 0.05), and there was no difference of daily values of manufactory control among viscometer C and D (F = 0.33,3. 14, P > 0.05), but significant daily difference existed when control A was tested by viscometer C and D (F = 5.76, 8.00, P < 0.05). Under SR of 30 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers(F =0.31, 0.18, 2.26, P >0.05), and there was no difference of daily values of control A among 3 viscometers' (F = 1.03, 1.83, 2.40, P > 0.05); Under SR of 200 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers (F =2.59, 0.68, 2.96, P > 0.05), and there was no difference of daily values of control A among 3 viscometers (F=2.31, 3.01, 2.28, P>0.05). When control A was tested under SR of 1 s~(-1) in 49 laboratories nationwide, the WBV values in groups of viscometer B, C and D were (18.47±1.30), (11.17±2.38), viscometer D and C were 63.75% and 21.3%. Conclusions Control A could fully mimic the properties of whole blood steadily on viscometer B, but partially mimic viscometer C and D, so the control A is most appropriate for viscometer B. Because current non-Newtonian fluid internal quality control could mimic rheological properties of whole blood under specifically conditions, laboratories should evaluate the consistent degree between control and whole blood, only the candidates which can mimic the properties of whole blood approximately could be chosen as quality control of WBV. When third-party control is chosen to be samples of EQA, its applicability should be in consideration. Pretest should be performed adequately to define applicability of third-party control, so as to reduce the difference among laboratories due to applicability of control and reflect detection quality of laboratories exactly.

Objective??To?evaluate?the?effects?of?Icariin?(ICA)?on?the?proliferation?and?osteogenic?differentiation?of?human?periodontal?ligament?stem?cells?(hPDLSCs)?in vitro and?in vivo. Methods??An?enzymatic?digestion?block?was?used?in vitro?to?culture?hPDLSCs,?which?were?separated?and?purified?by?limited?dilution?cloning.?The?hPDLSCs?were?identified?using?cell-surface?markers?and?cocultured?with?1×10?7 mol·L?1 ICA?solution.?The?proliferation?ability?of?these?cells?was?determined?by?thiazolyl?blue?tetrazolium?bromide?(MTT)?assay.?After?staining?with?alkaline?phosphatase?(ALP),?osteogenesis?was?detected?by?enzyme-linked?immunosorbent?assay.?Osteoblast-related?genes?were?analyzed?by?reverse?transcription-polymerase?chain?reaction.?Alizarin?red?staining?was?performed?to?measure?the?level?of?calcium?deposition.?The?hPDLSCs?were?cocultured?with?1×10?7 mol·L?1 ICA?and?nano-hydroxyapatite?scaffolds?in vivo?before?transplantation?into?subcutaneous?tissues?of?nude?mice.?Osteogenic?abilities?were?histochemically?analyzed?after?30?days?of?induction.?Results??The?hPDLSCs?were?affected?by?1×10?7 mol·L?1 ICA,?and?MTT?assay?showed?that?the?proliferation?of?the?groups?treated?with?ICA?in vitro?was?better?than?that?of?the?control?groups?on?the?second?day.?The?ALP?activity?of?the?treated?hPDLSCs?was?significantly?enhanced?after?cell?culture?for?3,?5,?and?7?days.?The?gene?expression?of?osteoblastic?markers?was?also?significantly?enhanced?after?7?days.?The?deposition?of?mineralization?after?incubation?with?1×10?7 mol·L?1?ICA?increased?compared?with?the?control?after?cell?culture?for?14,?21,?and?28?days.?Furthermore,?the?bone?expression?of?the?treatment?groups?in vivo?was?significantly?enhanced?com-pared?with?that?of?the?control?groups.?Conclusion??Treatment?with?1×10?7 mol·L?1 ICA?can?significantly?promote?proliferation?and?differentiation?of?hPDLSCs?in vitro?and?in vivo.?ICA?can?effectively?function?as?a?bioactive?growth?factor?in?periodontal?tissue?engineering?to?replace?traditional?growth?factors.

[Objective] To explore the optimal pressure parameters for chyle plasma filtration under low-temperature conditions, and to improve the quality of chyle plasma treatment and filtration efficiency by improving experimental methods. [Methods] The filtration efficiency and filtration time of 30 severe chyle plasma samples under conventional preparation environment pressure and under preparation environment with a controlled filtration membrane pressure difference of 0.5 bar were compared. [Results] The absorbance of severe chyle plasma samples before and after filtration under two different preparation pressures was statistically significant (P<0.05), and both achieved the expected filtration effect. Under the preparation environment of controlling the pressure difference of the filtration membrane to 0.5 bar, the filtration was faster and with better effect, which was statistically significant compared to the conventional preparation environment pressure (P<0.05). [Conclusion] By selecting the optimal pressure parameters for filtering chyle plasma under low-temperature conditions, the efficiency of chyle plasma filtration under low-temperature conditions has been improved, and the practicality and reliability of low-temperature filtration technology have been enhanced.