This study aimed at preparing the chemical reference substance of garcinol.A preparative high-speed countercurrent chromatography (HSCCC) was adopted for the isolation of garcinol from the twigs of Garcinia multifiora Champ,G.esculenta Y.H.Li and the fruits of G.xanthochymus Hook.f.ex T.Anders.The crude extracts were separated by HSCCC with two phase solvent systems composed of n-hexane-ethyl acetate-95％ ethanol-water (8∶8∶12∶4,volume ratio) using the upper stationary phase and the lower mobile phase.Under the conditions of a flow rate of 2.0 and 4.0 mL· min-1,and the apparatus rotation at 850 rpm,the column temperature at 25℃ and the detection wavelength at 254 nm,the fraction containing garcinol was purified by the Sephadex LH20 chromatography.The purity test in the extracts was determined by high-performance liquid chromatography.As a result,the contents of garcinol were 45,281 and 4080mg respectively separated from 104.765 g twigs of G.multiflora Champ,105.270 g G.esculenta Y.H.Li and 102.318 g the fruits of G.xanthochymus Hook.f.ex T.with the purity of 98.72％,98.36％ and 98.42％.Compared with the sample pretreatments and separation efficiency of the three plants,it was found that the fat-soluble extracts rapidly and efficiently extracted using n-hexane-ethyl acetate-95％ ethanol-water (5∶5∶5∶5,volume ratio) contained abundant garcinol taking fruits of G.xanthochymus Hook.f.ex T as the raw material with the combination of HSCCC and Sephadex LH-20 chromatography.In conclusion,it was indicated that the combination method is efficient with high operability and large preparation quantity.

The classical prescriptions in Typhoid Miscellaneous Disease can be used to alleviate the common hung cancer symptoms and complications like fever, cough, pain and pleural effusion effectively. The preoperative medication can improve lung cancer patients’ surgical tolerance, and postoperative medication can facilitate patients’ postoperative rehabilitation. And if combined with chemotherapy, they cound reduce toxicity and enhance efficiency, enhance the patients’ immunity and improve their life quality. Research shows that classical prescriptions can improve the immunity of lung cancer model mice, facilitate apoptosis of tumor cell and control its migration. This paper sumed up the clinical and experimental research of lung cancer treatment with classical prescriptions in recent 5 years.

Clinical data of 4 cases of infants with hemophilia complicated with hematorrhachis diagnosed in Guangzhou Women and Children′s Medical Center from January 2017 to December 2018 was retrospectively analyzed.The clinical manifestations of 4 children was atypical, only showing irritability and crying, poor spirit and weak head in the early stage, and torticollis and limb weakness in the late stage may easily contribute to a misdiagnosis.The spinal magnetic resonance imaging is an important diagnosis of hematorrhachis.The most common site of disease was cervicothoracic and thoracolumbar spinal epidural hemorrhage.Of the 4 patients in this article, 1 patient was treated with surgery combined with infusion factor Ⅷ replacement therapy.Three patients were treated with infusion factor Ⅷ replacement therapy alone to obtain clinical rehabilitation.The prognosis of early diagnosis was good, the time from onset to diagnosis in 3 cases was within 1 week, after treatment all had rehabilitation; 1 case within 2 weeks of diagnosis, partial rehabilitation.It is suggested that hemophilia complicated with hematorrhachis in infants is hidden, and it can only be irritable and crying in the early stage, and neurological symptoms appear in the late stage.Improving diagnostic awareness, early diagnosis and early infusion factor replacement therapy are critical for prognosis.

Objective To explore the mechanism of the nerve growth factor (NGF) accommodated by electro-acupuncture in rats with diabetes peripheral neuropathy (DPN).MethodsThe rat DPN model was established with streptozotocin (STZ). The rats were randomly divided into the normal group, model group, western medicine group and electro-acupuncture group. The animals of the electrol-acupuncture group were treated with electrol-acupuncture on "Zusanli" and "Shenshu" after DPN model established. The nervous tissues of all animals were examined with the techniques of immunity and fluorescent quantitation PCR to detect the albumen and mRNA expression of NGF.ResultsThe albumen and mRNA expression of NGF of the animals in the model group, western medicine group and electro-acupuncture group were significantly lower than that in the normal group ( P<0.05～0.01), and the albumen and mRNA expression of NGF was up-regulation in the electro-acupuncture group after electro-acupuncture treatment and was significantly higher than that in the model group ( P<0.01).ConclusionElectro-acupuncture on "Zusanli" and "Shenshu" can up-regulate the albumen and mRNA expression of NGF, and improve reparation of the sciatic nerve in DPN animals.

Objective To compare the effect of glucosamine (Virtral-s) on the cartilage oligomeric matrix protein (COMP) secretion of chondrocytes and synoviocytes in vitro.Methods Chondrocytes and synoviocytes isolated from knee cartilage of osteoarthritic patients were cultured by phased enzymatic digestion.Sera containing Virtral-s of the experimental animals were obtained after orally administrated Virtral-s at the dosages that equal to human.Cells were cultured in the medium with Virtral-s containing sera.Super-natant COMP level was tested by enzyme-linked immunoabsorbent assays (ELISA).Results COMP conceu-tration of synoviocytes cultured in vitro was significantly higher than that of chondrocytes (P＜0.05).Virtral-s could significantly increase COMP secretion in cultured chondrocytes in vitro (P＜0.05),however,it had a weaker role on synoviocytes,ie,it could only mildly reduce COMP secretion of synoviocytes.Conclusion Glucosamine (Virtral-s)-containing serum can promote COMP secretion of chondrocytes in vitro,and it has no significant effect on synoviocytes in vitro.

BACKGROUND: Pathophysiological mechanism of local microenvironment is complex after central nerve injury; especially,both inflammatory reaction at an acute phase and formation of secondary glial scar have tremendous effects on effective regeneration of axon, regeneration and arrangement of local nerve cells, proliferation and migration of local stem cells;therefore, it becomes a basic reason for blocking nerve repair in an early period. Thus, how to effectively resist inflammatory factors in injured region at an acute phase and how to optimize microenvironment of neural regeneration are the most important strategies for repairing spinal cord injury in recent years.OBJECTIVE: To design, establish and screen the best expression of interleukin-6 receptor (IL-6R) α to inhibit shRNA adenovirus expression vector by using spinal cord injury models.DESIGN: Duplicative measurement study.SETTING: Department of Spine Surgery, the Second People's Hospital of Shenzhen.MATERIALS: A total of 40 healthy Wistar rats, either gender, 8-10 weeks old, were selected in this study. Rabbit-anti-rat glial fibrillary acidic protein (GFAP) antibody Ⅰ was provided by Santa Cruz Compan; siRNA eukaryon expression plasmid pGenesil (cohtaining green fluorescent expression system) was provided by Wuhan Jingsai Bioengineering Company.METHODS: The experiment was carried out in ImmuneOpening Laboratory, Basic Medical Faulty, Tongji Medical College, Huazhong University of Science and Technology, and Medica Laboratory Center, the Second People's Hospital of Shenzhen in November 2006. Three pairs of shRNA template which composed of 19 bp reverse repeated motif of IL-6 receptor (IL-6R) α target sequence with 9 bp spacer were designed and synthesized, then the recombinant adenovirus expression vectors with green fluorescence protein were constructed in vitro respectively. The acute spinal cord injury models were completed, and the adenovirus recombinants were regionally injected post 12 hours after spinal cord injury;in addition, the inhibitory effect of RNA interference (RNAi) on expression of IL-6R in local region after spinal cord injury were detected by using real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot so as to screen adenovirus expression vector which had the best inhibitory effect on expression of IL-6R.MAIN OUTCOME MEASURES: Inhibitory effect of RNAi on expressions of IL-6R RNA and protein in local region after spinal cord injury.RESULTS: Sequence analysis showed that IL-6R-shRNA recombinant adenovirus expression vector was successfully constructed, and optimal IL-6R-shRNA recombinant adenovirus expression vector was screened by using real-time fluorescence quantitative RT-PCR and Western blot. The IL-6R expressions were 49% and 56% at the levels of mRNA and protein, respectively.CONCLUSION: The IL-6R--shRNA recombinant adenovirus expression vector is successfully constructed and screened.The gene expression of IL-6R can be highly inhibited after acute spinal cord injury.

Objective To review the experience of treatment for prosthetic valve endocarditis(PVE)after mitral valve replacement(MVR)in 16 cases.Methods From September 1979 to December 2010,16 patients were diagnosed as PVE after MVR by modified Duke University diagnostic criteria.There were 10 males and 6 females.Their ages ranged from 19 to 55 years old(mean 28 years old).The incidence of PVE was 0.97％(16/1657)after MVR.Blood culture positive was in 5 cases.Medical treatment alone was performed in 10 patients who were treated by using Penicillin or Vancomycin with other broad-spectrum antibiotics,using Fluconazole and Amphotericin if necessary.Combined medical and surgical management were performed in 6 cases.One emergency case was operated because of obstinate heart failure.Five cases underwent operation after adequate antibiotics treatment and general condition improvement.The infective tissue and vegetation were aggressively debrided after the infective prosthetic valve removed.Before the new valve was replaced paravalvular tissue must be flushed with diluted iodine solution and large quantities of normal saline.Tricuspid valve repair (TVR)was performed during the same period in 3 cases.Results Hospital death occurred in 8 patients and only 2 patients were recovery in group with medical treatment only.The main reasons of death were infective shock and cardiac failure in 4 patients,cerebral complications(embolism or bleeding)and multiple organs failure in 4 cases.While 6 patients cured after combined medical and surgical management.Follow-up had been carried out in 8 cases for 1.7 to 15 years(mean 5.1 years).Eight years later one patient was re-operated because of severe tricuspid regurgitation and paravalvular leak.There was no PVE recurrence in others.Conclusion Combined medical and surgical management for PVE get a better result than medical treatment alone.Good prognosis lies in timely diagnosis,adequate antibiotics,aggressive debridement of infective tissue and better myocardium protection during operation.

Background Studies show that regulatory T cells (Treg) are a kind of T cell subsets to negatively regulate immune response,and play an important role in maintaining immune homeostasis and immune tolerance.Autoimmune uveitis is an autoimmune disease,the regulation of Treg cells in pathogenesis and progression of autoimmune uveitis is not fully unelucidated.Objective This study was to observe the dynamic changes of Treg in experimental autoimmune uveitis (EAU) rats and explore the role of Treg cells in the pathological process of EAUrats.Methods Eighty four 6-8 week-old SPF Lewis rats were randomly divided into model group and control group.The mixed emulsifier of interphotoreceptor retinoid-binding protein (IRBP)1177-1191,tuberculin (TB),complete Freund adjuvant (CFA) and PBS (300 μl) was subcutaneously injected in double rear foot pad,abdominal side and back,and only equal amount of TB,CFA and PBS emulsifier was used in the same way in the control group.Ocular inflammation symptoms was examined at 9,13,18,23,28,35 and 48 days after modeling and scored based on the severity of the inflammatory.Six rats of each group were sacrificed in above time points respectively for the histopathological examination of iris,ciliary body and retinas by haematoxylin-eosin staining.The lymphocytes were isolated and cultured from rat spleens,and the proportion of Foxp3-labelled cells,a specific marker of Treg cells,was assayed by flow cytometry.The relative expression level of Foxp3 mRNA in the lymphocytes detected by using realtime quantitative PCR (RT-PCR).The use and care of the rats complied with the ARVO Statement.Results Eye inflammatory response appeared at 8 days after immunization,showing vasodilation and hyperemia of rat iris in the model group,and the response peaked at 13 days,with exudation and hypopyon in the anterior chamber.The highest inflammatory scores were 3.75±0.42 at day 13,and the ocular inflammation reaction was gradually relieved after that and disappeared at 23 days after immunity.A significant difference in ocular inflammatory scores of model rats was found among different time points (F =81.709,P ＜ 0.001);while no inflammatory symptom was observed in the control group.Histopathology examination showed obvious infiltration of inflammatory cells in the iris,ciliary body and retinas in model rats,including neutrophils,lymphocytes and mononuclear cells.The proportion of Foxp3-labelled cells in spleen lymphocytes was (5.50 ± 0.64)％,(13.36 ± 0.98)％,(10.34 ± 0.79)％,(9.58 ± 1.02)％,(6.73 ±0.81)％ and (5.58 ± 0.47) ％ in the model group on day 13,18,23,28,35,48 respectively,with statistically significant differences in comparison with (2.80 ± 0.38) ％,(3.36 ± 0.53) ％,(3.65 ± 0.57) ％,(3.37 ± 0.43) ％,(3.33±0.50)％ and (3.13±0.61)％ in the control group (t=-6.272,-15.556,-11.910,-9.753,-6.154,-5.491,all at P＜0.01).The change trend of Foxp3 mRNA expression was consistent to the dynamic change of the proportion of Foxp3-labelled cells.Conclusions The pathogenesis and development is closely associated with the dynamic changes of CD4+ CD25 + Foxp3+ Treg cells in EAU rats.

Objective To investigate the effects of ulinastatin (UTI) combined with magnesium isoglycyrrhizinate (MgIG) on the expression of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) in lung tis?sue of rats with pulmonary fibrosis induced by bleomycin (BLM). Methods Ninety Wistar rats were randomly divided into five groups: BLM group, methylprednisolone (MTH) group, UTI group, MgIG group and UTI combined with MgIG (UTI+MgIG) group, n=18 for each group. The rat model of pulmonary fibrosis was established by injecting bleomycin through tra?chea in five groups. Twenty-four hours after treatment with BLM,rats were treated with normal saline every day in BLM group, and rats were treated by corresponding drugs in other groups. Six rats of each group were killed at the 7th,14th and 28th day respectively. The pathological changes of alveolitis and pulmonary fibrosis were evaluated by HE staining, and ex?pression levels of TGF-β1 and CTGF in lung tissues were detected by immunohistochemistry method. Results (1) Com?pared with BLM group, the degree of alveolitis and pulmonary fibrosis was reduced in other groups. There was significant dif?ference in alveolitis at the 7th and 14th day between UTI+MgIG group and BLM group. And there was significant difference in pulmonary fibrosis at the 14th and 28th day between UTI+MgIG group and BLM group (P<0.05). (2) Compared with BLM group, the expression levels of TGF-β1 and CTGF were decreased in other groups. In UTI+MgIG group, the expres?sion levels of TGF-β1 were significantly lower at the 7th and 14th day compared with those in UTI group and MgIG group, and which were significantly lower at the 28th day than those in MTH group, UTI group and MgIG group (P<0.05). The ex?pression levels of CTGF were significantly lower at the 7th day in UTI + MgIG group than those in UTI group and MgIG group, and which were significantly lower at the 14th and 28th day than those in MTH group, UTI group and MgIG group (P<0.05). Conclusion The combination of UTI and MgIG can alleviate alveolitis and fibrosis in BLM-induced pulmonary fibrosis rats, which might related with the down-regulation of TGF-β1 and CTGF expressions.

Objective To investigate the clinical features and the imaging examination changes of bacterial meningitis in children of different age groups,and to offer theoretical basis for the clinic diagnosis of bacterial meningi-tis. Methods The sick children with bacterial meningitis treated in Guangzhou Women and Children′s Medical Center from January 2011 to June 2013,were recruited and divided into three groups according to the age,including newborns group,infants group and more than 1 year group. Eighty-nine cases with purulent meningitis were recruited,included 58 males and 31 females. Among them,there were 34 cases in the newborns group,41 cases in the infants group and 14 ca-ses in the more than 1-year group. The information on the clinical features,laboratory examination and the imaging ex-amination were summarized and analyzed,and were compared among the 3 groups. Results (1)Among them,54 cases with high fever,44 cases with respiratory symptoms,12 cases with alimentary tract symptoms. (2)The major neurologi-cal features included convulsions(44 cases,49. 4%),fatigue(36 cases,40. 4%),vomiting(21 cases,23. 6%),cervical rigidity(9 cases,10. 1%),fontanel full(7 cases,7. 9%),headache(6 cases,6. 7%),limb paralysis(5 cases,5. 6%). The percentage of high fever,vomiting,headache,cervical rigidity in the more than 1 year group were significantly more than those of the newborns group and the infants group(χ2=10. 093,12. 063,34. 466,7. 177,all P<0. 05). (3)Among them,whitebloodcell(WBC)accountswerefrom2.20to60.60×109/L,themeanwas(16.49±10.37)×109/L.Hy-persensitive C-reaction protein concentration in blood was 4. 00 to 376. 53 g/L,the mean was (131. 07±86. 91) g/L. In cerebrospinal fluid(CSF),WBC accounts were from 1 to 21 800×106/L,the mean was (910. 05±274. 07)×106/L, the glucose concentration from 0. 00 to 4. 50 mmol/L,the mean (1. 72±1. 03)mmol/L,the protein concentration from 0. 42 to 4. 89 g/L,the mean was(1. 64±1. 03)g/L. In 40 cases with atypical CSF change, 23 cases with CSF glucose ratio(CSF glucose/blood glucose) less than or equal to 0. 4, and 15 cases with blood cultures positive. (4)The image examination showed magnetic resonance imaging( MRI) abnormalities in 51/75 cases,computerized tomography( CT) scan abnormalities in 15/30 cases. The percentage of convulsions,the brain MRI abnormalities and the MRI display rate of bacterial meningitis in the infants group were significantly more than those of the newborns group and the more than 1 year group(χ2=11. 768,9. 047,7. 674,all P<0. 05). The display rate of meningitis and subdural hydroma by the brain MRI were significantly more than those of the head CT scan(χ2=7. 430,5. 291,all P<0. 05). Conclusions Be-cause of the atypical clinical features of bacterial meningitis in newborn and infant, lumber puncture should be per-formed in all doubtful cases who had a fever and/or seizure. CSF glucose less than or equal to 0. 4 of simultaneously ob-tained blood glucose value,the enhanced MRI sequence or blood cultures were useful for the likelihood of meningitis,if CSF chemistries and cytology vary atypically. The MRI sequence can significantly mostly improve the display rate of bacterial meningitis than the enhance CT.