[Correlation between metabolic syndrome and prognosis of patients with clear cell renal cell carcinoma].

OBJECTIVE: To investigate the effects of MetS on the prognosis of patients with clear cell renal cell carcinoma (ccRCC). METHODS: Clinical and pathological data and the laboratory test of ccRCC 342 patients with diverticular stones who underwent ccRCC who underwent radical or partial nephrectomy were retrospectively collected and analyzed.The patients were divided into MetS group and non-MetS group, and the subgroups were defined according to the tumor size. The overall survival (OS), cancer-specific survival (CSS), and progression-free survival (PFS) of the two groups were analyzed by univariate Cox analysis, and the subgroup analyses were also performed. Kaplan-Meier survival curve and survival analysis for OS, CSS, and PFS of the two groups and the subgroups were conducted. RESULTS: Univariate Cox analysis showed that MetS was a protective factor of postoperative OS [hazard ratio (HR)=0.551, 95%CI: 0.321-0.949, P=0.031], CSS (HR=0.460, 95%CI: 0.234-0.905, P=0.025), and PFS (HR 0.585, 95%CI: 0.343-0.998, P=0.049) in the patients with ccRCC. In the subgroup with tumor size≤4 cm, MetS was not associated with postoperative OS (HR=0.857, 95%CI: 0.389-1.890, P=0.702), CSS (HR=1.129, 95%CI: 0.364-3.502, P=0.833), and PFS (HR=1.554, 95%CI: 0.625-3.864, P=0.343). In the subgroup with tumor size>4 cm, Mets was a protective factor of postoperative OS (HR=0.377, 95%CI: 0.175-0.812, P=0.013), CSS (HR=0.280, 95%CI: 0.113-0.690, P=0.006), and PFS (HR=0.332, 95%CI: 0.157-0.659, P=0.002); Obesity was a protective factor of postoperative CSS (HR=0.464, 95%CI: 0.219-0.981, P=0.044), and PFS (HR=0.445, 95%CI: 0.238-0.833, P=0.011). Kaplan-Meier survival analysis showed that the long-term survival of patients with MetS was better than those without MetS in OS (P=0.029), CSS (P=0.021), and PFS (P=0.046); for the subgroup with tumor size≤4 cm, there was no significant difference in postoperative OS (P=0.702), CSS (P=0.833), and PFS (P=0.339) between patients with and without MetS; For the subgroup with tumor size>4 cm, the OS (P=0.010), CSS (P=0.003), and PFS (P=0.001) of patients with MetS were better than those without MetS. CONCLUSION: MetS was a protective factor of postoperative OS, CSS, and PFS in the patients with ccRCC, which was more obvious in subgroup with tumor size>4 cm. And obesity, the component of MetS, was correlated with postoperative OS and CSS.

[Risk factors of persistent hypertension in primary aldosteronism patients after surgery].

OBJECTIVE: To analyze the risk factors of persistent hypertension in patients who underwent adrenalectomy for primary aldosteronism and to evaluate the predictive value of the aldosteronoma resolution score (ARS) scoring system for surgical outcomes of adrenalectomy for primary aldosteronism. METHODS: We reviewed the clinical characteristics of patients who underwent adrenalectomy for primary aldosteronism from 2018 to 2021 at Peking University People' s Hospital to recognize risk factors of uncured hypertension after surgery. Based on the patient' s clinical outcomes, the patients were divided into complete success group and partial/absent success group. Risk factors for persistent hypertension were analyzed. The value of the ARS scoring system was assessed by the area under the curve (AUC). RESULTS: In this study, 112 patients were included. Most of the patients benefited from the surgery for 94.6% were a complete or partial clinical success after follow-up for at least 6 months. According to postoperative hypertension status, the patients were divided into complete success group (51 cases) and partial/absent success group (61 cases). There were statistical differences between the two groups in age, body mass index (BMI), waist circumference, duration of hypertension, number of preoperative antihypertension medications, preoperative systolic blood pressure, history of diabetes, history of cardiovascular and cerebrovascular diseases, serum creatinine, estimated glomerular filtration rate(eGFR), high-density lipoprotein cholesterol and triglyceride. Logistic regression analysis showed that age (OR=1.111, 95%CI: 1.029-1.199), waist circumference (OR=1.073, 95%CI: 1.013-1.137), pre-operative systolic blood pressure (OR=1.033, 95%CI: 1.008-1.060) and history of cardiovascular and cerebrovascular diseases (OR=16.061, 95%CI: 1.312-196.612) were the risk factors for uncured hypertension in primary aldosteronism patients after surgery, but female gender not. The median ARS in the complete success group was 4 and in the partial/absent success group, it was 2. Among the patients with ARS of 4-5, the cure rate of hypertension was 76.5%. The area under the curve of ARS was 0.743. CONCLUSION: The history of cardiovascular and cerebrovascular diseases is a significant risk factor for persistent hypertension after surgery in primary aldosteronism patients. ARS scoring system has a certain value in predicting the postoperative hypertension status of primary aldosteronism patients. However, further research is still needed on a prediction model for surgical outcomes of primary aldosteronism which is more suitable for the Chinese population is still needed.

[Clinical and pathological analysis of small renal cell carcinoma].

OBJECTIVE: To analyze the clinical and pathological features of small renal cell carcinoma (RCC), especially of those with diameter less than 4 cm and to understand the characteristics and factors related to recurrence and progression. METHODS: A total of 200 patients with RCC were stratifiedly selected for retrospective analysis. Their baseline demographic features, tumor-specific clinical features, pathological features of renal lesions, especially microscopic features were collected. The patients were divided according to the largest diameter of renal tumor lesions. Univariate analysis was used to compare the differences between tumor staging and microscopic pathological features between the groups. Binary multivariate Logistic regression was used to investigate factors related to tumor progression and prognosis in the patients with small RCC. RESULTS: The tumor diameters of 127 RCC patients were less than 4 cm and most of them had clear cell renal cell carcinoma (ccRCC). The increase in tumor diameter resulted in significantly higher T stage (P<0.01), higher WHO/International Society of Urological Pathology (ISUP) grade (P<0.05) and increasing chance of lymph node metastasis (P<0.01). Even when the tumor diameter was less than 4 cm, the patients might still have perirenal fat invasion, renal sinus invasion and greater elevated tumor grade (greater than grade 3) and synchronous lung metastasis. The incidences of intravascular thrombus (9.3% vs. 0) and tumor necrosis (27.8% vs. 5.5%) in the patients with RCC between 4-7 cm were significantly higher than those with RCC less than 4 cm (P<0.01). Sub-group analysis of small RCC (less than 4 cm) indicated that the patients with RCC between 2-4 cm were more likely to have intratumoral hemorrhage (44.7% vs. 23%, P<0.05) and necrosis than those with RCC less than 2 cm (8.2% vs. 0, P=0.095). Logistic regression analysis of small RCC showed that the incidence of tumor invasion to renal capsule was higher in ccRCC (OR=5.15, 95%CI: 1.36-19.52). Necrosis was closely related to the formation of peritumor pseudocapsule in small RCC (OR=14.90, 95%CI: 1.41-157.50). Increase in the tumor diameter was related to higher tumor grade (greater than grade 3) (OR=3.49, 95%CI: 1.11- 10.93). CONCLUSION: The tumor stage and grade of small RCC (less than 4 cm) are low, but extra-renal invasion and synchronous distant metastasis may occur. Internal hemorrhage and necrosis in tumor, ccRCC subtype, along with microscopic features, such as the renal capsule invasion and perirenal pseudocapsule formation are relevant factors of malignant behavior of small RCC and could be considered in prognosis evaluation.

[A case of gastric duplication presenting as a retroperitoneal cystic lesion].

Gastric duplication is a rare congenital anomaly accounting to 4% of all gastrointestinal tract duplications. It is usually diagnosed pre-natally or during infancy. Retroperitoneal gastric duplication is very rare in adult patients. It is generally asymptomatic or presents with non-specific symptoms. The sensitivity of classical imaging modalities for retroperitoneal gastric duplication is weak. Retroperitoneal gastric duplication could be misdiagnosed as other retroperitoneal cystic lesions and it could be definitely diagnosed by typical findings of abdominal enhanced computed tomography (CT) and endoscopic ultrasonography (EUS) plus EUS:guided fine needle aspiration biopsy in gastric duplicated cyst. A female adult patient was diagnosed by an exceptional hospital CT as retroperitoneal cystic lesion and was admitted in our hospital in March 2019. The abdominal enhanced CT in our hospital preoperatively indicated a cystic lesion above the left kidney and the maximum cross section was 9.0 cm×5.2 cm. Lymphocysts and others should be identified and the lesion might be from the stomach. The retroperitoneal gastric duplication was not diagnosed before operation. In the operation laparoscopy revealed the cystic lesion in the retroperitoneum. The pancreas, left adrenal gland, and left kidney were compressed by the cystic lesion. The top of the lesion was connected with the small curvature of the stomach, but it was not communicated with the stomach. The retroperitoneal cystic lesion was completely excised and the surrounding organs were not damaged. The retroperitoneal gastric duplication was clearly diagnosed by the findings in the operation and the pathology of the lesion. The patient was discharged a week after surgery without gastrointestinal and pancreatic injuries. Therefore, the differential diagnosis of retroperitoneal cysts should consider the possibility of retroperitoneal gastric duplication. For patients with typical CT findings or unclear boundaries between the tumor and the stomach wall, gastric duplication could be diagnosed by EUS plus EUS:guided fine needle aspiration biopsy in the cyst. Retroperitoneal gastric duplication cyst could be completely excised by laparoscopy through abdominal cavity and the stomach wall could be completely repaired.

[Minimal fat renalangiomyolipoma with multiple lymph nodes enlargement and postoperative refractory lymphatic fistula: a case report and literature review].

Renal angiomyolipoma (AML) is a common benign tumor in the urinary system, mainly composed of adipose tissue, blood vessels and muscle tissue. Renal AML is sporadic in most of patients, while a few are associated with tuberous sclerosis. Classical renal AML occurs predominantly in middle-aged females. Most cases are found incidentally during imaging examinations. The fat content makes AML have unique imaging characteristics and is easy to be identified with other renal tumors. However, the amount of fat varies in each tumor. AML that contains only microscopically detectable fat and whose amount of intratumoral fat may be too small to be identified on unenhanced computed tomography (CT) images is termed minimal fat or fat-poor renal AML, which appears as a high density shadow in the renal parenchyma on unenhanced CT images. Thus, it can be difficult to distinguish it from renal cell carcinoma (RCC) on imaging. Since the imaging findings are atypical, the diagnosis depends on pathological results. In addition, a few of AML can mimic malignant neoplasms. Recent studies suggested that AML might involve to peri-renal or renal sinus fat, regional lymphatics and other visceral organs, as well as inferior vena cava, which further makes the diagnosis more difficult. However, there is currently no reports about involvement of regional limphatics in minimal fat renal AML. In this article, we report a 27-year-old female patient without family history of tuberous sclerosis, who came to visit the hematologist because a high density shadow near the left kidney was found during CT scan which was accompanied by neck, armpits, groin, abdominal cavity and retroperitoneal lymph nodes enlargement. She was suspected of lymphoma in the beginning and transferred to Department of Urology to perform laparoscopic left renal mass and retroperitoneal lymph node excision and pathological examination for a definitive diagnosis. Finally, pathologic results revealed AML. Postoperative continuous lymphatic fistula developed and the retroperitoneal drainage of chylous fluid was 100-200 mL per day, lasting for 12 weeks. The fistula was successfully closed after conservative treatment including fasting and rehydration. This article summaries and discusses the diagnosis and treatment of renal AML with lymph nodes enlargement and the management of postoperative refractory lymphatic fistula by reviewing the related cases and literature.

[Congenital renal arteriovenous fistula complicated with multiple renal arteries malformation: case analysis].

Congenital renal arteriovenous fistula complicated with multiple renal arteries malformation is rare and hard to diagnose at early stage. Blood loss and complications after embolization are both severe. Some cases can be diagnosed by ultrasound, enhanced CT scan or digital subtraction angiography (DSA). Cystoscopy and ureteroscopy can identify the location of bleeding, exclude tumors, and discharge ureteral obstruction. A case of congenital renal arteriovenous fistula complicated with multiple renal arteries malformation was reported to investigate the pathogenesis, clinical characteristics, diagnosis and treatment of congenital renal arteriovenous fistula with multiple renal arteries malformation. A 36-year-old female patient with congenital renal arteriovenous fistula with multiple renal arteries malformation was hospitalized in the Department of Urology of Peking University People's Hospital. Five days before admission, the patient experienced whole course painless gross hematuria for 5 days with many blood clots. The patient's blood pressure was 90/70 mmHg, and hemoglobin was 60 g/L. The urinary CT scan showed a right hydronephrosis associated with dilatation of the upper ureter which was obstructed by space occupying lesion of the lower ureter. Many clots in the bladder could also be found in the CT scan. Cystoscopy showed many blood clots in the bladder and confirmed that the bleeding was fromthe right ureteral orifice. Ureteroscopy confirmed that the bleeding was from the right renal pelvis and many blood clots in the right ureter, and found no tumor in the right ureter and renal pelvis. We cleared the blood clots in the right ureter and inserted a ureteral stent.We thought that renal vascular malformation of the right kidney might lead to the hematuria from right renal pelvis. DSA showed a double renal arteries malformation in the right kidney. The diagnosis of "renal arteriovenous fistula" was considered with renal arteriovenous fistula in the right kidney. Selective arteriography revealed the presence of tortuous, coiled, dilated, and multichannelled vessels in the middle of the right kidney. With stainless steel coils, we embolized the vessels which supplied the fistula. Four days after the procedure, gross hematuria disappeared. Five days after the procedure, the patient's anemia improvedand the patient was discharged in good condition. Four months after the procedure, gross hematuria did not recur. The Doppler showed that the right kidney was normal and the renal dynamic showed that the right kidney function was normal. So DSA is the golden standard for diagnosis of congenital renal arteriovenous fistula complicated with multiple renal arteries malformation. Confirming the number of renal arteries by abdominal aorta angiography is necessary to avoid missed diagnosis. Renal arterial embolization is safe and effective.

[Effect of luteal-phase gonadotropin-releasing hormone agonist administration on pregnancy outcome in IVF/ICSI cycles: a systematic review and Meta-analysis].

Objective: To evaluate the potential efficacy and safety of gonadotropin-releasing hormone agonist(GnRH-a) administration in the luteal-phase on in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) cycles in assisted reproductive technology (ART). Methods: The relevant papers published before November 2015 were electronically searched in PubMed, EMBASE, Cochrane Library, WHO ICTRP, ClinicalTrials.gov, CNKI, CBM and WanFang database to collect randomized controlled trial (RCT) involving GnRH-a administration in the luteal-phase on IVF/ICSI cycles in ART. Two reviewers independently screened literature according to the inclusion and exclusion criteria, extracted data, and assessed methodological quality according to the Cochrane Handbook. Then, Meta-analysis was performed using Stata 13.0. Results: A total of 3 406 patients, 3 280 IVF/ICSI cycles from 11 RCT were subjected to Meta-analysis. All cycles presented statistically significantly higher rates of live birth/ongoing pregnancy (RR=1.29, 95%CI: 1.11-1.51), clinical pregnancy (RR=1.24, 95%CI:1.08-1.43) and multiple pregnancy (RR=1.95, 95%CI:1.21-3.14) in patients who received luteal-phase GnRH-a administration compared with those who did not. Conclusions: These findings demonstrate that the luteal-phase GnRH-a administration could increase birth/ongoing pregnancy rate, clinical pregnancy rate and multiple pregnancy rate in all cycles, so it may be an ideal choice for luteal phase support in patients undergoing IVF/ICSI therapy.

[Analysis of metabolic factors and relevance with the grade and stage in patients with renal cell carcinoma].

OBJECTIVE: To investigate the association between metabolic factors, such as obesity, blood pressure, blood glucose, serum lipid profile, and the histopathological characteristics of renal cell carcinoma. METHODS: The medical records of 382 consecutive renal cell carcinoma patients who underwent radical or partial nephrectomy at Peking University People's Hospital from January 2009 to January 2015 were retrospectively reviewed. Metabolic factors were collected from the records, including weight, body mass index, waist circumstance, blood pressure, fasting blood glucose, serum total triglyceride, serum total cholesterol, serum low-density lipoprotein-cholesterol and serum high-density lipoprotein-cholesterol. The patients were divided into different groups according to tumor grade, stage and diameter. Statistics analysis, such as t test, Mann-Whitney U test and Logistic analysis, were performed to investigate the association between metabolic factors and grade, stage and tumor diameter of renal cell carcinoma. RESULTS: A total of 80 (20.94%) of the tumors were classified as high grade disease, 63 (16.49%) were classified as advanced disease and 153 (40.05%) tumor diameter more than 4 cm. The patients in high grade group were found to have lower high-density lipoprotein-cholesterol level than in low grade group (P=0.015), body mass index, total cholesterol and high-density lipoprotein-cholesterol were found to be lower in advanced disease than in localized disease (P=0.022, P=0.005 and P=0.006, respectively), and low-density lipoprotein-cholesterol was found to be lower in larger tumors (P=0.030). Other factors were comparable between the different groups. The results of Logistic analyses showed that, body mass index (OR=0.906, 95%CI: 0.852-0.986, P=0.023) and total cholesterol (OR=0.660, 95%CI: 0.492-0.884, P=0.005) were associated with the tumor stage, high-density lipoprotein- cholesterol level was significantly associated with tumor grade (OR=0.293, 95%CI: 0.108-0.797, P=0.016) and stage (OR=0.204, 95%CI: 0.065-0.635, P=0.006), and low-density lipoprotein-cholesterol level was significantly associated with tumor diameter (OR=0.756, 95%CI: 0.586-0.975, P=0.031). CONCLUSION: The results of our study indicate that metabolic factors, especially obesity and serum lipid profile, are closely related with the histopathological characteristics of renal cell carcinoma.

[Analysis of metabolic factors and relevance with the grade and stage in patients with renal cell carcinoma].

OBJECTIVE: To investigate the association between metabolic factors, such as obesity, blood pressure, blood glucose, serum lipid profile, and the histopathological characteristics of renal cell carcinoma. METHODS: The medical records of 382 consecutive renal cell carcinoma patients who underwent radical or partial nephrectomy at Peking University People's Hospital from January 2009 to January 2015 were retrospectively reviewed. Metabolic factors were collected from the records, including weight, body mass index, waist circumstance, blood pressure, fasting blood glucose, serum total triglyceride, serum total cholesterol, serum low-density lipoprotein-cholesterol and serum high-density lipoprotein-cholesterol. The patients were divided into different groups according to tumor grade, stage and diameter. Statistics analysis, such as t test, Mann-Whitney U test and Logistic analysis, were performed to investigate the association between metabolic factors and grade, stage and tumor diameter of renal cell carcinoma. RESULTS: A total of 80 (20.94%) of the tumors were classified as high grade disease, 63 (16.49%) were classified as advanced disease and 153 (40.05%) tumor diameter more than 4 cm. The patients in high grade group were found to have lower high-density lipoprotein-cholesterol level than in low grade group (P=0.015), body mass index, total cholesterol and high-density lipoprotein-cholesterol were found to be lower in advanced disease than in localized disease (P=0.022, P=0.005 and P=0.006, respectively), and low-density lipoprotein-cholesterol was found to be lower in larger tumors (P=0.030). Other factors were comparable between the different groups. The results of Logistic analyses showed that, body mass index (OR=0.906, 95%CI: 0.852-0.986, P=0.023) and total cholesterol (OR=0.660, 95%CI: 0.492-0.884, P=0.005) were associated with the tumor stage, high-density lipoprotein- cholesterol level was significantly associated with tumor grade (OR=0.293, 95%CI: 0.108-0.797, P=0.016) and stage (OR=0.204, 95%CI: 0.065-0.635, P=0.006), and low-density lipoprotein-cholesterol level was significantly associated with tumor diameter (OR=0.756, 95%CI: 0.586-0.975, P=0.031). CONCLUSION: The results of our study indicate that metabolic factors, especially obesity and serum lipid profile, are closely related with the histopathological characteristics of renal cell carcinoma.

Serotonin regulates osteoblast proliferation and function in vitro

The monoamine serotonin (5-hydroxytryptamine, 5-HT), a well-known neurotransmitter, also has important functions outside the central nervous system. The objective of this study was to investigate the role of 5-HT in the proliferation, differentiation, and function of osteoblasts in vitro. We treated rat primary calvarial osteoblasts with various concentrations of 5-HT (1 nM to 10 µM) and assessed the rate of osteoblast proliferation, expression levels of osteoblast-specific proteins and genes, and the ability to form mineralized nodules. Next, we detected which 5-HT receptor subtypes were expressed in rat osteoblasts at different stages of osteoblast differentiation. We found that 5-HT could inhibit osteoblast proliferation, differentiation, and mineralization at low concentrations, but this inhibitory effect was mitigated at relatively high concentrations. Six of the 5-HT receptor subtypes (5-HT1A, 5-HT1B, 5-HT1D, 5-HT2A, 5-HT2B, and 5-HT2C) were found to exist in rat osteoblasts. Of these, 5-HT2A and 5-HT1B receptors had the highest expression levels, at both early and late stages of differentiation. Our results indicated that 5-HT can regulate osteoblast proliferation and function in vitro.

Abnormal lipid metabolism down-regulates adenosine triphosphate synthase ß-subunit protein expression in corpus cavernosum smooth muscle in vitro and in vivo.

Metabolic syndrome is closely related to erectile dysfunction (ED), and hyperlipidaemia is considered a major risk factor for ED. Adenosine triphosphate (ATP) synthase is believed to play an important role in metabolic syndrome; it has been hypothesised that ATP synthase contributes to ED development. We have verified this hypothesis using primary cultured human corpus cavernosum smooth muscle (HCCSM) cells treated with excessive free fat acid (FFA) and a high-fat diet (HFD) mouse model. Our results showed that high fatty factors could cause lipid accumulation in HCCSM cells, which could result in abnormal lipid metabolism, such as high levels of triglycerides, cholesterol and glucose in the HFD mice. There was a remarkable down-regulation of ATP synthase and p-Akt after in vivo and in vitro excessive FFA treatments. These results indicated that abnormal lipid metabolism could induce ATP synthase down-regulation via the Akt phosphorylation pathway and that ATP synthase may be a target of lipotoxicity in corpus cavernosum smooth muscle cells.

Selective resection of dorsal nerves of penis for premature ejaculation.

Premature ejaculation (PE) is one of the most prevalent male sexual dysfunctions. Selective resection of the dorsal nerve (SRDN) of penis has recently been used for the treatment of PE and has shown some efficacy. To further clarify the efficacy and safety of SRDN on PE, we performed a preliminary, randomized, placebo-controlled clinical observational study. Persons with the complaints of rapid ejaculation, asking for circumcision because of redundant foreskin, intravaginal ejaculation latency time (IELT) within 2 min, not responding to antidepressant medication or disliking oral medication were randomly enrolled in two groups. From April 2007 to August 2010, a total of 101 eligible persons were enrolled, 40 of them received SRDN which dorsal nerves of the penis were selectively resected, and those (n = 61) enrolled in the control group were circumcised only. IELT and the Brief Male Sexual Function Inventory (BMSFI) questionnaire were implemented pre- and post-operatively for the evaluation of the effect and safety of the surgery. There are no statistically significant differences in the baseline data including mean ages, mean IELTs, perceived control abilities and the BMSFI mean scores between the two groups. With regard to the post-operative data of the surgery, both IELTs and perceived control abilities were significantly increased after SRDN (1.1 ± 0.9 min vs. 3.8 ± 3.1 min for pre- and post-operative IELT, respectively, p < 0.01),whereas the post-operative results were not significantly improved for the control group (1.2 ± 0.7 min vs. 1.5 ± 1.1 min, p > 0.05). Also, there were no statistically significant differences both in BMSFI composite and subscale scores between the two groups after surgery. Hence, we conclude that SRDN is effective in delaying ejaculation and improving ejaculatory control, whereas erectile function is not affected. The results imply that SRDN may be an alternative method for the treatment of PE for some patients.

A mutation in the Kit gene leads to novel gonadal phenotypes in both heterozygous and homozygous mice.

The direct sequencing of the Kit cDNA obtained from mutant mice was used to reveal the molecular nature of the W(-3Bao) ENU-induced mutation. There was a T to A transversion at the 441st nucleotide in the W(-3Bao) open reading frame (ORF), which introduced a pre-mature termination codon at residue 147. The gross embryonic development, hematopoiesis and spermatogenesis were examined in the mutant mice. There was no visible difference among the W(-3Bao/+), W(-3Bao/3Bao) and wild type embryos before embryonic day 12.5. W(-3Bao/3Bao) embryos appeared pale after E14.5 and dwarf after E16.5. An extremely low level of hematochrome and large red blood cells were found in W(-3Bao/3Bao) 18.5 days old embryos, leading to the stillbirth of the homozygotes. In 18.5 days old embryos the spermatogonia of W(-3Bao/3Bao) embryos did not migrate to the contorted seminiferous tubules properly, but instead were found in the interstitial tissue. The spermatogonia of W(-3Bao/+) or W(+/+) mice were present in both the interstitial tissue and contorted seminiferous tubules. In the adult male hetereozygotes, there are contorted seminiferous tubules with no spermatogonia, suggesting that the migration defect was dominant. In female W(-3Bao/3Bao) ovaries, primordial follicles were absent while primordial follicles appeared clearly in the ovaries of W(-3Bao/+) or W(+/+) mice. With a nonsense mutation in the Kit gene, W(-3Bao/+) mice show white spotting and an abnormal development of the contorted seminiferous tubules and W(-3Bao/3Bao) mice are stillborn due to severe macrocytic anemia, and have abnormal genital glands in both the male and female.

Relevance between striatal expression of Fos, proenkephalin mRNA, prodynorphin mRNA and rotation induced by l-stepholidine in 6-hydroxydopamine-lesioned rats.

AIM: To study that l-stepholidine (SPD) regulates the expression of proenkephalin (PENK) and prodynorphin (PDYN) mRNA and Fos in the striatum after rotational test in the 6-hydroxydopamine (6-OHDA)-lesioned rats. METHODS: PENK and PDYN mRNA levels were examined with in situ hybridization, and Fos expression was detected with immunocytochemistry. The data were semi-quantified with image analyzer. RESULTS: (1) Following repeated SPD treatment, the rotation was kept on high activity in the 6-OHDA-lesioned rats. (2) SPD significantly elicited Fos expression in both sides of striatum, particularly in the denervated one. Repeated administration of SPD, Fos expression declined on both sides, particularly in the intact one. (3) In the denervated striatum of 6-OHDA-lesioned rats, the PENK mRNA level was extremely increased vs that in the intact striatum. This high level of PENK mRNA was significantly reduced by 7-d treatments of SPD. SPD also reduced the level of PENK mRNA in the intact striatum. However, the level of PDYN mRNA did not show significant change in both sides of striatum after denervation or SPD treatment. CONCLUSION: In the 6-OHDA-lesioned rats, the rotation induced by SPD was kept on a high activity, which was in pace with the inducement of Fos expression and the reduction of expression of PENK mRNA in the denervated striatum. But then the lesion and SPD treatment had no remarkable effect on the expression of PDYN mRNA.

Oral administration of doxycycline reduces collagenase and gelatinase activities in extracts of human osteoarthritic cartilage.

OBJECTIVE: To determine whether oral administration of doxycycline in clinically relevant doses will suppress activities of collagenase and gelatinase in extracts of human osteoarthritic (OA) cartilage. METHODS: Femoral heads were obtained from 21 patients undergoing arthroplasty for endstage hip OA. Activities of collagenase and gelatinase were measured in extracts of the OA cartilage from patients who received doxycycline, 100 mg bid or qam for 5 days before surgery (n = 5 and n = 6, respectively), 200 mg as a single dose 3 days before surgery (n = 4); or no doxycycline (n = 6). RESULTS: Five days of doxycycline treatment, in a dose of either 100 mg bid or 100 mg qam, inhibited gelatinase activity in the cartilage extracts (p = 0.003, 0.008, respectively). The bid dose also inhibited collagenase activity (p = 0.002), but inhibition of collagenase with 100 mg qam did not quite reach statistical significance (p = 0.055), in comparison with the values for the untreated OA controls. The single 200 mg dose, given 3 days before procurement of the cartilage, was ineffective in inhibiting metalloproteinase activity. CONCLUSION: Oral administration of doxycycline significantly inhibited collagenase and gelatinase activity in human OA cartilage. The effective dose is likely to be well tolerated during chronic administration, e.g., in a clinical trial to assess the potential of the drug to modify cartilage breakdown in OA.

The role of the membrane-spanning and extra-membranous regions of the iron-sulfur protein in its assembly into the cytochrome bc1 complex of yeast mitochondria.

The assembly of six deletion mutants of the Rieske iron-sulfur protein into the cytochrome bc1 complex was investigated by immunoprecipitation from detergent-solubilized mitochondria with specific antisera against either the iron-sulfur protein or the intact cytochrome bc1 complex. After import, the mutant proteins lacking residues 41-55 or 66-78, located at the membrane-spanning region of the protein, and residues 182-196 located at the C-terminus of the protein, were assembled in vitro into the bc1 complex approximately 50% as effectively as the wild type iron-sulfur protein suggesting that these regions of the iron-sulfur protein may not be critical for the assembly. By contrast, only trace amounts of the mutant proteins lacking residues 80-95, 122-135, 138-153 located in the extra-membranous region of the iron-sulfur protein were assembled into the bc1 complex. After import in vitro into mitochondria isolated from a cytochrome b-deficient yeast strain, the mutants lacking residues 41-55 and 182-196 were assembled as efficiently as the wild type; however, the mutants lacking residues 55-66 and 66-78 were assembled less efficiently in the absence of cytochrome b suggesting that the hydrophobic membrane-spanning region, residues 55-78, of the iron-sulfur protein, may interact with cytochrome b during the assembly of the bc1 complex.

Effects of oral doxycycline administration on histomorphometry and dynamics of subchondral bone in a canine model of osteoarthritis.

OBJECTIVE: To determine whether oral administration of doxycycline, in a dose that inhibits cartilage breakdown in a canine cruciate deficiency model of osteoarthritis (OA), affected formation or resorption of subchondral bone. METHODS: Ten healthy adult mongrel dogs underwent transection of the left anterior cruciate ligament. Five were given doxycycline, 3.5 mg/kg/day, orally, from the day after surgery until they were sacrificed 6 weeks later; the other 5 dogs served as positive OA controls. Three weeks before sacrifice, each dog was given an intravenous injection of the fluorochrome label, calcein, on each of 2 consecutive days; 10 days later, the injection regimen was repeated. At sacrifice, samples of bone from the medial femoral condyle of both knees of each dog were obtained for staining with Villanueva tetrachrome to localize the fluorochrome label and McNeal's tetrachrome for measurement of osteoid and cell counts. RESULTS: Cruciate deficiency resulted in a marked decrease in bone mass, with increased osteoclastic activity and increased bone formation. Doxycycline treatment did not significantly affect either bone formation or bone resorption. CONCLUSION: Doxycycline protects against joint breakdown in this OA model via inhibition of matrix metalloproteinases in articular cartilage, rather than through an effect on subchondral bone.

Proton NMR investigation of the oxidized three-iron clusters in the ferredoxins from the hyperthermophilic archae Pyrococcus furiosus and Thermococcus litoralis.

The 3Fe forms of ferredoxins (Fd) from the hyperthermophilic archaebacteria Pyrococcus furiosus (Pf) and Thermococcus litoralis (Tl) have been investigated by 1H NMR. A combination of one-dimensional nuclear Overhauser and two-dimensional NOESY and bond correlation spectroscopy provides the assignment of the aromatic residues, one conserved valine, and the location of the signals for each of the three cysteines coordinated to the clusters. Dipolar contacts between the Trp 2 and Tyr 46 in Pf Fd and from an invariant phenylalanine to an invariant valine and a cluster cysteine in both Fd confirm a folding pattern for these proteins that is very similar to that of the crystallographically characterized ferredoxin from the mesophile Desulfovibro gigas. The sequence-specific assignment of the buried cysteine near the invariant phenylalanine has been made. The temperature dependence of the contact-shifted cysteinyl residues reveals a distinct 2:1 asymmetry in the magnetic coupling among the three high-spin ferric ions, in that one cysteine exhibits Curie behavior, while the other two cysteines display anti-Curie behavior. These magnetic properties are rationalized qualitatively on the basis of a magnetic coupling scheme where two iron couple to yield an intermediate spin of 2 which couples to the remaining S = 5/2 iron to yield the total cluster spin 1/2. This magnetic asymmetry appears to be a characteristic feature of oxidized 3 Fe clusters. Pf Fd also undergoes a dynamic equilibrium between two alternate forms that differ slightly in the environment of two of the coordinated cysteines. Analysis of the pattern of the contact shifts for the three cysteines in the two ferredoxins suggests that the cysteine coordinated to the unique iron does not have the same sequence origin.

Doxycycline inhibits type XI collagenolytic activity of extracts from human osteoarthritic cartilage and of gelatinase.

Doxycycline was shown to inhibit digestion of exogenous type XI collagen by homogenates of human osteoarthritic cartilage in vitro. On SDS-PAGE, the cleavage products generated by cartilage extracts were larger and less abundant, indicating less complete cleavage, when doxycycline (10 or 30 microM) was added to the samples. The inhibitory effect was concentration dependent. Purified gelatinase from canine kidney epithelial cells, which also digests type XI collagen, was inhibited in a similar manner by doxycycline. If tetracyclines inhibit this metalloproteinase activity in articular cartilage in vivo, they could modify cartilage breakdown in osteoarthritis.