[Objective]To sum up pro. Lian Jianwei’s clinical experience in treating acne. [Method] By proved recipe analysis, it shows pro. Lian Jianwei ’s treatment based on differentiation of signs from different angles and the clinical thought of flexibly applying ancient formulae on acne. [Result] Pro. Lian Jianwei is good at revising ancient formulae for treating acne with good effect, according to patients ’pulse signs and mechanism;if the food is stagnated in the gastric-intestine, which wil transform into heat and up along the channel, Baohe Pil shal be used to remove stagnation and heat; if the liver is de-pressed with heat, deficient blood and weak spleen, Xiaoyan Powder shal be used for regulation and stretching the wood; use Wenqing Drink to nourish blood and remove toxin for deficient blood and hot liver, toxic heat implicated in spleen and stomach; apply Zisheng Pil to regulate and tonify middle part of the body for deficient spleen and stomach with turbidness and dyspepsia, etc. [Conclusion] Pro. Lian Jianwei has rich clinical experience, his clinical thought of “treatment based on differentiation of signs, with flexible methods and formulae, coping with shifting diseases by sticking to a fundamental principle”, worthy our deep experiencing and study.

Purpose] This paper aims to summarize the clinical practice of using Buzhong Yiqi Decoction to treat the obstructed nine orifices. [Approach] After explaining Li Dongyuan’s theory that those obstructed nine orifices reflecting headaches and tinnitus are caused by intestines and stomach diseases,which is in his Suwen On Deficiency and Excess and analyzing several simple cases of using buzhong yiqi decoction to treat the obstructed nine orifices,this paper tries to trace the theoretical origin of Prof. Lian’s practice and present the practical experience of flexible usage of Buzhong Yiqi Decoction. [Result] The outcome of several patients under treatment shows that the short-and-mid-term treatment is very effective. [Conclusion] Prof. Lian’s philosophy of traditional Chinese medicine reflects his attitude that “those patients suffering from insufficiency of primordial Qi and clear Yang failing to distribute”or“clear Yang coming out of stomach”ask for strengthening the spleen and stomach so their stomach recovers to the healthy state. As a result,their Yang Qi has been greatly improved,the spleen and stomach in the middle energizer have returned to their original place,reaching the autonomous regulation of the nine orifices,which has shed some light on the usefulness of Buzhong Yiqi on treatment diseases other than those of spleen and stomach.

Objective To evaluate the value of MSCTA in combination with CTV in diagnosis of pulmonary embolism and deep venous thrombosis (DVT) of lower limb. Methods 47 patients with pulmonary embolism proved clinically were examined with MSCTA using 120 kV,210 mA,collimater of 0.75 mm and the automatic bolus traching technique,the speed of contrast agent infu-sion was 3～4 m/s after 170 s delayed,venous scan of lower limbs was performed. Results 47 cases were well perform the trunk of bilateral pulmonary artery, pulmonary lobar artery, pulmonary segemental artery, pulmonary inferior segmental artery could be showed by MSCTA, MPR, MIP and VRT. 363 arterial segments were involved and 31 cases accompanied with DVT. Conclusion MSCTA in combination with CTV is of significant value in detecting pulmonary embolism accompanied with deep venous thrombo-sis.

BACKGROUND:Orthopedists should pay more attentions to nonunion prevention in view of nonunion treatment, that is, active interventions should be taken to avoid bone delayed union and nonunion.
OBJECTIVE:To explore the effect of composite tissue-engineered scaffold constructed by nano-hydroxyapatite/polyamide 66 (nHA/PA 66) combined with bone marrow mesenchymal stem cels to repair a femoral fracture with severe nonunion.
METHODS:Rat bone marrow mesenchymal stem cels were isolated and culturedin vitro, and then they were divided into three groups: bone marrow mesenchymal stem cels without osteogenic induction, with osteogenic induction or combined with nHA/PA 66 folowed by osteogenic induction as control group, test group or composite group, respectively. Then osteogenic differentiation of bone marrow mesenchymal stem celsin vitrowas analyzed by measuring alkaline phosphatase activity and alizarin red staining, cel adhesion on the nHA/PA 66 was observed using scanning electron microscopy, and the celgrowth and proliferation were detected by MTT assay. In the meanwhile, established Sprague-Dawley rat femur nonunion models were randomly divided into three groups: the areas of nonunion were implanted with nothing as blank control group,those were with nHA/PA 66 as simple scaffold group, and the others were with nHA/PA 66 combined with bone marrow mesenchymal stem cels as composite scaffold group. Afterwards, X-ray examination, micro-CT and Masson staining were used to evaluate the femoral healing.
RESULTS AND CONCLUSION:At 6 and 12 days after osteogenic induction, alkaline phosphatase activity in the test group was significantly higher than that in the control group; at 14 days, compared with the control group, the amount of mineralized nodules in the test group was significantly higher, which indicated that bone marrow mesenchymal stem cels after osteogenic induction could differentiate into osteoblasts. Attached cels spread wel on the scaffold with good proliferation activity, suggesting that nHA/PA 66 is suitable for cel adherence, proliferation and osteogenic differentiation. Besides, at 12 weeks after modeling, in the blank control group, no calus appeared in the nonunion region. In the simple scaffold group, the broken femur did not heal at 8 and 12 weeks after surgery. In the composite scaffold group, the broken femur did not heal at 8 weeks, but a lot of calus appeared; at 12 weeks, bone healing achieved and the scaffold was encased and absorbed.These findings demonstrate that the tissue-engineered bone scaffolds constructed by bone marrow mesenchymal stem cels and nHA/PA 66 effectively prevent bone nonunion by accelerating femoral healing in a rat femur nonunion model.

Objective To explore the biocompatibility of nano-hydroxyapatite/polyamide 66 (nHA/PA66) with human bone mesenchymal stem cells (hBMSCs) after osteogenic induction.Methods After hBMSCs were isolated and cultured in vitro,the experiment was conducted in 3 groups.Group A were hBMSCs subjected to no osteogenic induction,group B hBMSCs subjected to osteogenic induction,and group C was the composite of nHA/PA66 with hBMSCs subjected to osteogenic induction.Adhesion of the cells onto the nHA/PA66 in group C was observed by electron microscope scanning.Growth and proliferation of the cells in groups B and C were detected by MTI test at 1,2 and 3 weeks.The ability of osteogenic differentiation of hBMSCs in vitro was analyzed by alkaline phosphatase (ALP) activity and alizarin red staining.The ability of osteogenic differentiation of hBMSCs on nHA/PA66 was tested by ALP activity.Results Electron microscope scanning showed that the cells spread and attached well on the surface of the composite scaffold in group C;the proliferation of the cells in groups B and C showed no significant difference (P ＞ O.05).These suggested that the proliferation of hBMSCs was not affected by nHA/PA66.The number of mineralized nodules in group B was significantly larger than in group A (P ＜ O.05);the ALP activity of the cells in group A was significantly lower than in group B at 6 and 12 days (P ＜ 0.05);no significant differences were observed between groups B and C (P ＞ 0.05).These indicated that the hBMSCs were capable of osteogenic differentiation which was not affected by nHA/PA66.In groups B and C,the ALP activity of the cells at 12 days was significantly higher than at 6 days,indicating the ALP activity increased with increased induction time (P ＜ 0.05).Conclusion nHA/PA66 can be used as a carrier of hBMSCs in bone tissue engineering because hBMSCs can well adhere to,proliferate,and differentiate into bone on nHA/PA66 scaffolds.

Objective: To probe and reveal the brain tomic and the wise sharpening action of Fuyuanbunao Granules and Its toxicity and side effect. Methods: The effects of Fuyuanbunao Granules on the memory acquisition disturbance induced by pentobarbital sodium or anisodine, the memory consolidation disturbance induced by sodium nitrite, and the memory repetition disturbance induced by alcohol were observed. Meanwhile, the acute toxicity test was done. Results: The granules could obriously improve the memory disturbance induced by the factors obove, and the improvement effect varied with the dose. The effects of larger dose groups were more obvious. There were no death of animals or toxicity and sile effect in test. Conclusion:Fuyuanbunao Granules maybe have the cholinergic acction. It can promote the aerobic metabolism, improve the cerebral anoxia and the inhibited state of center nervous system. It has low toxicity and can be used in clinical application.

Objective:To observe the change of periphery and centra lymphocyte subsets at the crest-time of MOG_(35-55) induced EAE disease in mice,and to explore the alteration of cellular immunity and humoral immunity in the invasion process in EAE.Methods:MOG_(35-55) was used to establish EAE model in femina C57BL/6 mice.The behavioral changes and the histological scores were recorded after the mice were immuned .The changes of CD3~+CD4~+,CD3~+CD8~+,CD4~+CD25~+ and B220~+ on periphery and centra lymphocytes in spleen,brain and spinal cord were analyzed by flow cytometry.Results:The CD3~+CD4~+,CD3~+CD8~+,CD4~+CD25~+ and B220~+ lymphocytes were detected in the brain and spinal cord of EAE group mice,but they were not detected in CFA control group.The CD3~+CD4~+ and CD3+CD8+lymphocytes in the spleen of EAE crest-time group were lower than those in CFA control group(P＜0.05).The B220~+ lymphocytes were obviously higher than in the CFA control group (P＜0.01).And CD4~+CD25~+ lymphocytes were slight higher than the CFA control group.Conclusion:At the crest-time during EAE,the CD3~+CD4~+,CD3~+CD8~+lymphocytes of spleen reduced obviously,B220~+ lymphocytes increased markedly,and the CD4~+CD25~+ lymphocytes just have the increasing trend.It indicates that cellular immunity and humoral immunity coregulated the patho-process at the crest-time of EAE,T lymphocytes and B lymphocytes all played important roles in the pathogenesy of EAE.

Pig copper zinc superoxide dismutase (CuZnSOD) is an important antioxidant enzyme. Some studies focused on the function of CuZnSOD gene, but the transcriptional regulation of the CuZnSOD gene is not yet fully elucidated. Therefore, the aims of the study were to determine the core promoter region and to explore its mechanism of transcriptional regulation. The 853 bp DNA sequence of 5'-flanking promoter was amplified by performing PCR. A series of CuZnSOD promoter fragments with gradually truncated 5'-end were produced by nested PCR and inserted into pGL3-Basic vector. The activities of the promoters were measured by the dual-luciferase assay system after transient transfection into the NIH/3T3 cells. The results demonstrated that there were 2 potential transcription start sites in the regions from initiation codon to -87 bp and -266 bp, respectively. The region from -383 bp to +67 bp in CuZnSOD gene promoter showed higher activity than other regions, and further deletion analysis demonstrated that the region from -75 bp to -32 bp contained an essential promoter sequence for pig CuZnSOD gene transcription. In addition, several potential transcription factor binding sites were predicted with bioinformatics method. These results suggest that these transcription factor binding sites may be involved in the transcriptional regulation of CuZnSOD gene.
Animals ; Cloning, Molecular ; Gene Expression Regulation ; Genetic Vectors ; Mice ; NIH 3T3 Cells ; Promoter Regions, Genetic ; Superoxide Dismutase ; genetics ; Swine ; Transfection

BACKGROUND:Unlike the ilium derived from the paraxial mesoderm, the mandible from cranial neural crest has a unique mechanism. Core binding factorα1 (Cbfα1) is a key transcription factor for skeletogenic process. However, the role of Cbfα1/p56 subtype in mandible tissue is yet not clear. OBJECTIVE:To research the expression of Cbfα1/p56 subtype in bone marrow mesenchymal cells from rat mandible in vitro. METHODS:Bone marrow mesenchymal stem cells from rat mandible and ilium were in vitro isolated and purified by primary culture. The characteristics of bone marrow mesenchymal cells were compared through the methods of enzyme linked immunosorbent assay and real-time PCR, including growth curve, alkaline phosphatase activity and relative mRNA expression of Cbfα1 subtypes. RESULTS AND CONCLUSION:Bone marrow mesenchymal cells from rat mandible and ilium were successful y obtained. Bone marrow mesenchymal cells from the mandible proliferated more rapidly, alkaline phosphatase activity of which was higher than iliac cells. The relative mRNA expression of Cbfα1/p56 subtype in bone marrow mesenchymal cells from the mandible was more than that in iliac cells at 6 days of culture (P<0.05), while the expression of Cbfα1/p57 in each time showed no statistical significance (P>0.05). The results showed that Cbfα1/p56 is very significant in the early osteogenic differentiation of bone marrow mesenchymal cells from the mandible.

Objective The aim of this study was to investigate the relationship between phosphotyrosine interac-tion domain containing 1 ( PID1 ) gene and variation in intramuscular fat ( IMF ) content and the possibility to generate transgenic animals by testicular injection .Methods Expression vector pIRES2-acGFP-PID1 carrying pig PID1 gene was incubated with transfection reagents and injected into the testes of male New Zealand rabbits .We examined the F1 genera-tion by fluorescence detection , PCR, Western blotting and measuring the IMF content .The F1 generation gave birth to the F2 generation.Then we examined the F2 generation through detecting the positive rate and the IMF content .Results The exogenous PID1 gene and fluorescent protein gene were expressed at different levels both in the F 1 generation and the F2 generation, and the positive rates were 35.88%and 34.33%, respectively.The IMF content was significantly elevated (P<0.05) in the transgenic positive individuals compared with the negative ones and the control group , and the PID1 protein expression was similarly higher .Conclusions The results of this study demonstrate that PID 1 gene affects intramuscular fat content significantly .Moreover, the results of our analysis provide further evidence that transgenic animals can be gener -ated by testicular injection , and the exogenous gene can be inherited steadily .