BACKGROUND: Although cultured technique and purified method of Schwann cells (SCs) are changed successively,purity is still limited. Whether double negative selections can culture high-purified SCs still needs a further study.OBJECTIVE: To investigate the effects of double negative selections on growth and purity of SCs.DESIGN: Controlled animal study.SETTING: Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University,and the Beijing Institution of Otolaryngology.MATERIALS: The experiment was carried out in the Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University and the Beijing Institution of Otolaryngology between May 2006 and July 2006. Twenty new-born Wistar rats of 3 days (without gender limitation) were provided by the Experimental Animal Center of the Chinese Medical Academy.METHODS: The sciatic nerves of rats were digested by 0.2% collagenase. ① Grouping: All the rats were divided into 4 groups with 5 each. The experimental group was depurated by differential adhesion and Ara-c for 30 minutes. SCs in the control group 1 and 2 were managed by differential adhesion and Ara-c for 30 minutes, respectively. Cells in the control group 3 were not purified. ② Evaluation: The cell growth was observed under invert microscope; growth curves were measured with MTT method in the experimental group and control group 3 to observe cell proliferation. In addition, the SCs were identified by immunocytochemistry; the culture purity was calculated (ratio of S100 positive cells in each sight).MAIN OUTCOME MEASURES: ① Growth of SCs; ② immunocytochemistry stain of SCs; ③ growth curve of SCs; ④ purity of SCs.RESULTS: ① Growth of SCs: At 5 days after incubation, the SCs proliferated well. SCs were spindle, bipolar and sometimes tripolar, with obvious nuclei and a little cytoplasm. There was more fibroblast cell in the group 3 than that in experimental group. ② Growth curve: The SCs in experimental group entered into the logarithm multiplication period at the third day. ③ Immunocytochemistry: The positive-stained SCs were brown in its body and process, and arranged like swirl. The FB with spread and anomaly shape was slight blue. ④ The ratio of anti-S100 positive cells was higher in experimental group than that in group 3 (P ＜ 0.01).CONCLUSION: With the double negative selection, high-purified SCs are obtained effectively.

Objective To study the feasibility and clinical results of C2, 3 pedicle-lateral mass plate fixation in the treatment of unstable Hangman's fractures. Methods From March 1999 to July 2005, 25 cases of unstable Hangman's fracture were treated surgically using the C2, 3 pedicle-lateral mass plate. All the cases were followed up for the healing of bone and function of the cervical vertebrate. Results The average follow-up period was 38 months. Twenty-three cases had a perfect reduction. Three cases had kyphosis deformity. There were no other complications related to the operation. AH the cases had a perfect early fusion of the C2 pedicle isthmus and long-term stability of C2, 3 intervertebral space. The clinic results were good. Conclusion Since the C2, 3 pedicle-lateral mass fixation can provide immediate reduction and satisfactory biomechanical stability, it is a good device for unstable Hangman s fractures.

The suppressive effect of anti-KDR antibody against VEGF on proliferation of hemangioma-derived vascular endothelial cells (HVECs) was investigated. HVECs from one case of hemangioma in proliferative phase were cultured. Both primary culture and sub-culture were conducted in M199 medium. The HVECs of passage 3 were divided into 4 groups based on the concentrations of anti-KDR antibody. Cell count was performed and inhibitory rate of HVECs was measured before and 9 days after interference. The results showed that the number of HVECs in the anti-KDR antibody-treated groups was significantly decreased and the inhibitory rate of HVECs by anti-KDR antibody (50, 10 and 2 microg/mL) was 84%, 63% and 39% respectively at 9th day after interference, with the difference being significant. In the control group, the number of HVECs was increased significantly. In was concluded that the anti-KDR antibody could suppress the activity of VEGF through blocking the KDR, indicating the potential clinical applications of anti-KDR antibody in the treatment of hemangioma.
Antibodies/*pharmacology ; Cell Proliferation/drug effects ; Endothelium, Vascular/*pathology ; Hemangioma/*pathology ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor Receptor-2/*immunology

To study whether the development of hypertensive disorder complicating pregnancy is associated with -308G-->A, -850C-->T mutation at promoter of TNF-alpha gene, the -308G-->A, -850C-->T polymorphism was examined in patients and healthy pregnant women by PCR-RFLP technique. The frequencies of genotype and allele were compared between the two groups. The results showed that with -308G-->A polymorphism distribution, the allele frequency of TNF2 and the frequency of the genotype TNF2/1 in the patient group was significantly higher in the patient group than in control group (P<0.05). A significant difference in genotype distribution of -850C-->T polymorphism was observed between the two groups. The allele frequencies of T in patient group was higher in the control group as compared with the patient group. The frequencies of CT and TT genotypes were lower in the patient group. It is concluded that the TNF2 allele of -308 is associated with the occurrence of hypertensive disorder complicating pregnancy, while T allele of -850 may be the protective factor against the development of the disease. TNF2/1 CC may be susceptibility genotype of hypertensive disorder complicating pregnancy.
Case-Control Studies ; Gene Frequency ; Genotype ; Hypertension, Pregnancy-Induced/*genetics ; Mutation/*genetics ; Polymorphism, Genetic/*genetics ; Promoter Regions, Genetic ; Tumor Necrosis Factor-alpha/*genetics ; Young Adult

ObjectiveTo study the protective effects of Qiqiong(QQJN) on focal cerebral ischemia/reperfusion injury and its mechanism. MethodsMiddle cerebral artery occlusion(MCAO) was used to make focal cerebral ischemia/reperfusion model by intravascular nylon filament occlusion. The protective effects of QQJN were evaluated by investigating neurological function score, percentage of cerebral infarction, pathomorphology of brain, the activity of SOD and the content of MDA in hrain tissue,thrombogenesis and platelet aggregation in vitro. ResultsCompared with model group, QQJN(4.4、8.8g/kg)could decrease the neurological score in 8 and 22h after reperfusion, reduce the percentage of cerebral infauction,improve pathomorphology of brain, decrease the length, wet weight and dry weight of thromb and inhibit platelet aggregation. ConclusionQQJN had protective effects on focal cerebral ischemia/reperfusion injury. The role of anti-injury of free radicals,inhibit thrombogenesis and platelet aggregation should contribute to its neuroprotective effects.

The clinical efficacy and safety of topical propranolol hydrochloride gel in the treatment of superficial infantile hemangiomas (IHs) were assessed. Fifty-one cases of IHs from Oct. 2010 to Sept. 2011 were subjected to the topical propranolol hydrochloride gel intervention in Fuzhou General Hospital of Nanjing Military Commands, China. Changes in size, texture, color, peak systolic velocity of the hemangiomas, resistance index and adverse effects were observed. The results were evaluated by using Achauer system, and responses of IHs to pranpronolol were considered scale II (poor) in 4 patients (17.24%), scale II (moderate) in 18 patients (24.14%), scale III (good) in 22 patients (44.83%) and scale IV (excellent) in 7 patients (13.79%). The response of superficial hemangiomas was significantly better than other hemangiomas (P<0.05), and no differences in response were found among different primary sites (P>0.05). Our study indicates that topical application of 3% propranolol hydrochloride gel is effective and safe in treating IHs.

Objective:This study aimed to identify the morphology of mast cells by using a modified toluidine blue staining scheme,so as to provide a powerful reference for the experimental basis research of mast cells.Methods:Bone marrow-derived mast cells were induced in vitro.After 4 weeks,the cells were collected,fixed,and stained.Mast cells were fixed at different temperature during different time.The optimum condition was determined by comparing the effects of toluidine blue staining.Results:Bone marrow cells were induced to differentiate into mast cells by SCF and IL-3 in vitro.When mast cells were stained with modified toluidine blue staining,the staining effect was better.Mast cells were round or oval and the cell membrane was complete and the cytoplasm was filled with a large number of purple particles.Conclusion:In this study,we successfully applied a modified toluidine blue staining method to mast cells cultured in vitro.The results showed that the condition at 37 ℃ full fixation with staining could reduce the degeneration of mast cells.This method was easy to operate with good stability.It was suitable for the morphological observation of mast cells cultured in vitro.

OBJECTIVETo compare the therapeutic effect of Astragalus and Angelica Mixture (AAM) on treating CKD patients according to different CKD primary diseases, staging and TCM syndromes.

METHODSA multicentre, open-label, and self control clinical design was used, and thirty-two patients in line with inclusive criteria were recruited. Based on maintaining their previous basic CKD treatment, patients additionally took AAM (Astragalus and Angelica each 30 g), once a day, three months consisted of one therapeutic course. Serum creatinine (SCr), estimated glomerular filtration rate (eG- FR), 24 h urinary total protein (UTP), plasma albumin (ALB), hemoglobin (Hb), and changes of TCM syndrome factor integrals were compared before treatment, at the end of month 1, 2, and 3. The differences in the aforesaid indices were compared between CKD patients with different CKD primary diseases (chronic glomerulonephritis, chronic renal tubulointerstitial disease, hypertensive renal damage), different CKD stages (CKD 3 and CKD 4), and patients of qi-blood deficiency syndrome (QBDS) and non-QBDS.

RESULTSAAM could improve 78.12% (25/32) patients' renal function. Compared with before treatment, SCr decreased (12.08% +/- 10.11%), eGFR increased (21.14% +/- 18.55%), and ALB increased (2.76% +/- 1.97%) at the end of 3-month treatment (all P < 0.05). As for TCM syndrome factor integrals, compared with before treatment, the integrals for qi deficiency syndrome, blood deficiency syndrome, and yin deficiency syndrome decreased, while the integrals for dampness heat syndrome and turbid-toxin syndrome increased (all P < 0.05). There was no obvious difference in all indices except the integral for hypertensive renal damage patients of yin deficiency syndrome (P > 0.05). The SCr decreasing percent was 19.82% +/- 8.30% for patients of non-QBDS and 5.24% +/- 10.75% for patients of QBDS. The latter was higher with statistical difference (P < 0.05). As for TCM syndrome factor integrals, the integral differences of qi deficiency and blood deficiency were obviously higher in patients of QBDS, when compared with patients of non-QBDS (P < 0.05).

CONCLUSIONAAM could improve the renal function of CKD patients, elevate their ALB levels, and ameliorate associated qi deficiency syndrome, blood deficiency syndrome, and yin deficiency syndrome, especially for CKD patients of QBDS.

Adolescent ; Adult ; Aged ; Angelica ; Astragalus Plant ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phytotherapy ; Renal Insufficiency, Chronic ; drug therapy ; Treatment Outcome ; Yin Deficiency ; drug therapy ; Young Adult

Embryo Transfer ; Endometrium ; Female ; Fertilization in Vitro ; Humans ; Integrative Medicine

Because of hardness to heal and easiness to recurrence,chronic ulcer of lower limp has become one of the hardest diseases in clinic,which brings physical and mental pain to the patients.Although medical technology develops rapidly,to find a simple,effective and economic method is still the focus.Here,progress and current situation of treatment were summarized for chronic ulcer of lower limp.