Animals ; Antigens, CD ; metabolism ; Cell Movement ; Cell Proliferation ; Endoglin ; Genetic Therapy ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Neoplasms ; pathology ; therapy ; Receptors, Cell Surface ; metabolism ; Thy-1 Antigens ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

Diabetic peripheral neuropathy (DPN), one of the most common chronic complications of diabetes, is characterized by allodynia, hyperalgesia and spontaneous pain. Chinese epidemiological studies have shown that at least 25% diabetic patients suffered from painful DPN, which compromises patients' daily functioning and becomes a major health care problem. Although the pathogenesis of painful DPN is not fully understood and current treatment options are very limited, research in the field has advanced our understanding on the mechanism of painful DPN in the past Decade of Pain Research and Control. This review will mainly focus on evaluation of current diabetic animal models, possible molecular pathways and available therapies, with an emphasis on roles of purinergic receptor and its signaling transduction pathways. Common therapies address one or two DPN symptoms, while others offer wider symptom control, presumably by targeting pathophysiological mechanisms of DPN. Purinergic receptor signaling transduction pathways might become potential targets for treatment for painful DPN.
Animals ; Diabetes Mellitus ; physiopathology ; Diabetic Neuropathies ; physiopathology ; Humans ; Hyperalgesia ; physiopathology ; Pain ; physiopathology ; Receptors, Purinergic P2X ; physiology

Thirty healthy women and 101 patients with polycystic ovary syndrome (PCOS) were recruited. According to serum testosterone (T) level and homeostasis model assessment for insulin resistance (HOMA-IR) ,the correlation of T and body mass index (BMI) with insulin resistance was analyzed. The results showed that there were 39. 8％ normal,24. 5％ overweight,and 35.7％ obese among 101 PCOS patients. However,there were no significantly differences in BMI, fasting plasma glucose ( FPG ), triglyceride ( TG ), total cholesterol ( TC), low-density lipoprotein-cholesterol ( LDL-C), high-density lipoprotein-cholesterol ( HDL-C ), and HOMA-IR levels between PCOS patients with hyperandrogenemia ( T ≥ 0. 51 μg/L) and normal androgenemia ( T ＜ 0. 51 μg/L). BMI, FPG, TG, TC, and LDL-C levels were higher and HDL-C level was lower in patients with insulin resistance( HOMA-IR ≥ 2. 29 ) than in patients without insulin resistance ( HOMA-IR ＜ 2. 29, P＜0. 05 or P＜ 0. 01 ). Serum T levels were not significantly different between two groups. HOMA-IR was significantly correlated with BMI(P＜0. 01 ), not with serum T, suggesting that the gain of body weight is correlated with insulin resistance independent of serum T level.

A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.

Objective To improve the diagnosis and treatment of ureteral cancer. Methods A retrospective analysis of 24 cases of primary ureteral cancer treated from January 1990 to March 2005 was performed.The diagnostic value of ultrasound,IVU,CT,MRU and the patients' outcomes were reviewed. There were 19 males and 5 females aged 38-72 years(mean,59 years).The tumors were on the left side in 16 cases and on the right in 8.Of the 24 cases,17(71%)had gross hematuria and 7(29%)had micro- scopic hematuria.Urine cytology was performed in 16 cases with a positive rate of 6.3%.B-ultrasonic exami- nation showed hydronephrosis in 19 cases(79%)and low-echo space-occupying disease of middle-inferior ureter in 3(12%).IVU demonstrated hydronephrosis in 20 cases(83%)and filling defect of the diseased ureter in 3(12%).Retrograde pyelography showed filling defect of the diseased ureter in 16(76%)of 21 cases(5 cases had failure of intubation).CT scan was performed in 20 cases,indicating thickening of the ureteral wall and infiltration of the cancer in 14(70%).In 3 cases who had undergone spiral CT thin layer scan and 1 of 3 cases who had undergone MRU,the definite diagnosis was made.Results All the 24 pa- tients underwent surgical treatment.Among them,nephroureterectomy and bladder cuff or partial resection were performed in 18 cases,and nephrectomy and partial ureterectomy in 6 cases.Postoperative pathology showed transitional cell carcinoma in 23 cases,and adenoma in 1.Of the 14 cases during 1990-1999 peri- od,1,5,3,2,2 and 1 cases had survival time of 1,2,3,4,5 and 6 years,respectively.Of the 10 cases during 2000-2005 period,3 were lost to follow-up;2 survived for 3 years and 2,for 1 year;the other 3 who have survived near 5 years have been followed till now.Conclusions IVU and retrograde urography are the most common diagnostic measures for primary ureteral cancer.They can be used in combination with other imaging study to reduce missed diagnosis rate.The 5-year survival rate was lower because of late pathologic stage of the tumors in the patients of this series.

Objective To establish a method for isolation and cultivation of human invasive pituitary adenoma-derived fibroblast in vitro and explore their biological properties in order to investigate the role of the fibroblasts in the invasive growth of the pituitary adenoma.Methods The fibroblasts were isolated from hu- man invasive pituitary adenoma tissue by different rate of adhesion.Primary culture and passage culture were carried out,and growth characteristics of fibroblasts were observed under phase-contrast microscope in primary and passage culture.Immunohistochemical technique was used to identify the fibroblasts.The cell growth curve was measured by MTT method.The ultrastructure was observed under electronmicroscope.Results The cultured invasive pituitary adenoma-derived fibroblasts showed active proliferative ability.The cell bodies was bigger than other tissue fibroblasts,and the cell shape was irregular.The rough endoplasmic reticulum, Golgi complex and ribosome were profuse in the cytoplasm.The positive expression rate of collagen[and vi- mentin of the fibroblasts were above 95%.Conclusion The method of different rate of adhesion is success- fully utilized to culture human invasive pituitary adenoma-derived fibroblasts.These cells showed active prolif- erative ability,and maybe have an importent function in the invasive growth of the pituitary adenoma.

Objective To investigate the effect of local injection of Botulinum toxin type A(BTXA) on spasticity and function of the affected upper limb in stroke patients.Methods A total of 32 stroke patients were re- cruited and randomly divided into two groups:a BTXA group and a control group.All the patients had spasticity of upper limb muscles,which scored grade 2 to 3 with the Modified Ashworth Scale(MAS) ,and decreased elbow joint range of motion.The 16 patients in the BTXA group received BTXA injection in the biceps brachii muscles and flexor muscles of forearm on 10～15 points,while those in the control group did not.All the patients in both groups were treated with rehabilitation training techniques.The MAS,Fugl-Meyer upper limb function assessment and Barthel In- dex were employed to evaluate the changes of muscle tone,upper limb function and activity of living (ADL)perform- ance of the patients before injection and at 1st,2nd,6th 12th weeks after injection.Results The therapeutic effect between the BTXA group anti control group was significantly different in terms of biceps muscle tone,the scores of Fugl-Meyer upper limb function assessment and Barthel Index.Compared with preinjection,muscle tone was de- creased significantly and ADL performance was improved after injection in BTXA group.The effects of BTXA lasted more than 12 weeks.Conclusion Intramuscular muhipoint injection of BTXA was useful in reducing muscle spas- ticity,and was helpful for increasing motor ability of the affected upper limb and ADL performance of the stroke pa- tients.

Objective To investigate the clinical features,treatment response and prognosis in children with Takayasu′s arteritis(TA) in order to improve the understanding of TA.Methods A retrospective study of 10 children with TA was performed.All of them were admitted and diagnosed in Peking University First Hospital from Jan.1998 to Oct.2008.The clinical features,laboratory tests,imaging modalities,treatment response and prognosis were all collected and evaluated.Results There were 3 boys and 7 girls in the 10 patients with TA,and the ratio of male to female was 12.3.The onset was from 4 months to 9 years old,with average age at 5.5 years old.The average duration of diagnosis was 7.6 months.The incidences of hypertension,vascular bruits,albuminuria,convulsion were present in 100%,100%,70% and 40%,respectively.The clinical types included typeⅡ(60%),type Ⅲ(10%) and type Ⅳ(30%).The acute phase inflammatory indices of activity such as erythrocyte sedimentation rate(ESR),C-reactive protein(CRP) and white blood cell(WBC) were not evidently increased.Tuberculosis infection was found in 6 out of 10 patients and anti-tuberculosis treatment was performed.Six patients were treated with steroids and 3 cases of them were also given immunosuppressives cyclophosphamide or methotrexate.Three of the 10 patients received anti-hypertensive and vasodilator.Two patients received percutaneous translurminal angioplasty and 1 patient received nephrectomy.One patient died of renal failure,heart failure and shock.Conclusions The patients with TA had high prevalence of tuberculosis infection,diagnosis as often late because of lack of specific clinical features at the acute inflammatory period.When organic ischaemia occurred,treatment response was usually unsatisfactory.Patients with multi-systemic and multi-viscera lesions should have comprehensive examination,especially for those with hypertension,pulseless and vascular bruits,in order to rule out TA.Early ultrasonography,computed tomography and magnetic resonnance image methods are valued in eariler diagnosis and they are the key factors to improve prognosis.

Objective To study the different expressions of receptor activator of nuclear factor-kappa B ligang(RANKL) mRNA in spleens of rats fed with diet of low calcium and high fluoride. Methods A 2× 2×2 factorial design was used and the factors were calcium, fluoride and action time. In the design, 40 Wistar rats [average body mass(118.9±13.5)g] were divided into four groups randomly by weight: control with normal diet (0.790%, calcium), low calcium group with low calcium intake(0.063%, calcium), high fluoride group with normal diet and high fluoride intake(100 mg/L, fluoride) and low calcium and high fluoride group with low calcium and high fluoride intake. After 4 and 8 months, 5 rats of each group were sacrificed and total RNA was extracted from spleen. And the expression levels of RANKL mRNA were determined by reverse transcription polymerase chain reaction (RT-PCR). Results At time of 4 months, the expression level of RANKL mRNA was 0.13± 0.05,0.13± 0.03,0.17±0.02,0.27± 0.05 and at time of 8 months, it was 0.11 ± 0.01,0.16 ± 0.02,0.16± 0.03,0.36 ± 0.07 in control group, low calcium group, high fluoride group, low calcium with high fluoride group, repectively. The factorial design AVONA showed that low calcium and high fluoride had significant effects on RANKL mRNA expression(F = 40.224,56.679, all P < 0.05) while action time had not(F = 2.850, P > 0.05 ). The interactions of low calcium with high fluoride or high fluoride with action time were signifieant(F = 7.247, 18.789, all P < 0.05) while the interaction of high fluoride with action time was not(F = 1.751, P > 0.05). Conclusions Low calcium alone or high fluoride alone or low calcium with high fluoride or low calcium with action time can increase the the RANKL mRNA expression level. High fluoride does not affect the RANKL mRNA level as the action time is prolonged.

Objective To explore the potential mechanism of adrenal androgen excess in patients with polycystic ovary syndrome (PCOS), ACTH stimulation test was conducted and the polymorphisms in the promoter region of CYP21 A2 gene were screened to verify the variations related to the responsiveness to ACTH stimulation. Methods 30 healthy women and 101 PCOS patients, matched for age, were recruited from Ruijin hospital. Blood biochemical examinations were taken and sex hormone profiles obtained at baseline. 17 hydroxyprogesterone( 17OHP)was measured at 0 and 60 min in an ACTH stimulation test. The -710 bp -1 bp of the promoter region of CYP21A2 was sequenced in 87 PCOS patients and 30 control subjects. Results According to the post-stimulation 17 OHP levels obtained from 30 healthy women,PCOS patients were allocated into one group with high responsiveness to ACTH ( HR-PCOS, n = 21) and the other with normal responsiveness to ACTH ( NR-PCOS, n = 80). Compared with NR-PCOS subjects, HR-PCOS patients had higher testosterone( P<0.05), basal and post-stimulation 17OHP (both P<0.01)and dehydroepiandrosterone sulfate levels (P<0.05 or P<0.01) .whereas serum cortisol and androstenedione levels were not significantly different before and after ACTH stimulation test. The genotypes of locus -535 were well correlated with post-stimulation 17OHP levels (r = 0. 20,P = 0. 03) in PCOS patients and the control subjects. The genotype T/T or allele T was significantly more frequent in subjects with a higher fertile of post-stimulation 17OHP (P<0.05 or P<0. 01). The odds ratio(OR)for higher responsiveness to ACTH in women with allele T at -535 was 3. 69 (95% CI 1. 69-8. 06,P = 0. 000 7). Conclusions The PCOS patients with higher responsiveness to ACTH are characterized by severe hyperandrogenemia and adrenal androgenexcess,suggesting that adrenal androgen excess in some PCOS patients may be due to higher responsiveness to ACTH. The polymorphism of -535C>T in the promoter region of CYP21 A2 may play a role in regulating 21 hydroxylase gene expression and further influencing 17OHP responsiveness to ACTH.