A stereological observation of nuclear volumes and diameters of hippocampal neurons in aging rats / 解剖学报

Objective To determine if there is atrophy of neuronal nuclei in the aging rat hippocampus.Methods The entire hippocampal formation in a cerebral hemisphere (on a random side) was removed from normal male SD rats aged 1, 2 and 3 years (each age-group 5 rats), serial sections (section thickness 40 |xm) were cut after embedding in a resin of glycol methacrylate, and 3-4 sections were systematically sampled and stained with periodic acid, Schiff' s reagent and hematoxylin. The nuclear sizes of hippocampal neurons were estimated with stereological methods, first, the nuclei were sampled in a uniform random way using the optical disector; then, a Feret diameter of the largest nuclear profile at its focal plane (optical section) was measured as the nuclear diameter, and the nuclear volume was measured at the clearest focal plane of the nucleolus with the nucleator. Results The mean diameters and volumes of the neuronal nuclei in the whole hippocampus were 8.7-9.2 |xm and 392-457 (Jim, respectively, in the 3 age-groups, without significant statistical difference between groups. There were no significant difference between groups, either, in the nuclear diameter or volume in each hippocampal region——the GA (hippocampus proper), dentate gyrus or subiculum. Conclusion In general, no age-related changes have been observed in the nuclear sizes of neurons in the whole hippocampus in the rat from middle to old age.

Autoimmune enteropathy and primary biliary cholangitis after proctocolectomy for ulcerative colitis: A case report and review of the literature.

BACKGROUND: Autoimmune enteropathy (AIE) and primary biliary cholangitis (PBC) are both immune-mediated diseases. AIE or PBC complicated with ulcerative colitis (UC) are rare. There are no cases of AIE and PBC diagnosed after proctocolectomy for UC reported before, and the pathogenesis of these comorbidities has not been revealed. CASE SUMMARY: A middle-aged woman diagnosed with UC underwent subtotal colectomy and ileostomy due to the steroid-resistant refractory disease, and a restorative proctectomy with ileal pouch-anal anastomosis and proximal neoileostomy was postponed due to active residual rectal inflammation in January 2016. A few months after the neoileostomy, she began to suffer from recurrent episodes of watery diarrhea. She was diagnosed with postcolectomy enteritis and stoma closure acquired a good therapeutic effect. However, her symptoms of diarrhea relapsed in 2019, with different histological features of endoscopic biopsies compared with 2016, which showed apoptotic bodies, a lack of goblet and Paneth cells, and villous blunting. A diagnosis of AIE was established, and the patient's stool volume decreased dramatically with the treatment of methylprednisolone 60 mg/d for 1 wk and tacrolimus 3 mg/d for 4 d. Meanwhile, her constantly evaluated cholestatic enzymes and high titers of antimitochondrial antibodies indicated the diagnosis of PBC, and treatment with ursodeoxycholic acid (16 mg/kg per day) achieved satisfactory results. CONCLUSION: Some immune-mediated diseases may be promoted by operation due to microbial alterations in UC patients. Continuous follow-up is essential for UC patients with postoperative complications.

Clinical evaluation of traditional Chinese medicine on mild active ulcerative colitis: A multi-center, randomized, double-blind, controlled trial.

INTRODUCTION: Ulcerative colitis (UC) is a chronic inflammatory bowel disease (IBD) characterized by a relapsing-remitting course owing to recurrent intestinal inflammation. UC often has symptoms such as intermittent rectal bleeding, diarrhea, and abdominal pain. As the precise etiology of UC has not completely clarified, UC has become a public health challenge worldwide. According to an epidemiological survey, there were about 350,000 new cases of IBD in China from 2005 to 2014. By 2025, the number of IBD patients in China will reach 1.5 million. Traditional Chinese medicine (TCM) has been widely used to treat UC in China, however, it is still challenging to systematically determine the efficacy of in UC. Therefore, this trial aims to evaluate the clinical efficacy and safety of CHM in the treatment of mild active UC patients. METHODS: A multi-center, double-blinding, double-dummy, active-controlled, randomized trial will be established. A total of 240 patients in 6 centers with mild active UC (Mayo score is 3-5 points) and TCM syndrome of damp-heat stasis blocking and spleen-qi deficiency will be randomly allocated in the ratio of 1:1 to 2 groups: the experimental group and the control group. The experimental group will receive Hudi enteric-coated capsules (HEC) and enteric-coated mesalazine tablets placebo; the control group will receive enteric-coated mesalazine tablets and HEC placebo. Each group will be treated for 8 weeks. The primary therapeutic outcome: the rate of clinical efficacy and clinical remission at 8 weeks of treatment (last survey point) according to the modified Mayo score. The secondary outcomes: individual symptom score, TCM syndrome score, endoscopic response rate, mucosal healing rate, and quality of life scale score. Outcomes will be assessed at baseline and the end of the trial. Besides, intestinal mucosa, stools and blood biopsies from the mild active UC patients before and after treatment will be collected to reveal the underlying mechanisms. DISCUSSION: The results of this trial will provide compelling evidence of the efficacy and safety of HEC for treatment of mild active UC and preliminarily show the potential mechanism of how HEC acts. Finally, it will widen treatment options for patients with mild active UC.

Similarities and differences in clinical and pathologic features of inflammatory bowel disease-associated colorectal cancer in China and Canada.

BACKGROUND: Colorectal cancer (CRC) has become one of the major life-threatening complications in patients with inflammatory bowel disease (IBD), which includes ulcerative colitis (UC) and Crohn's disease (CD). This study aimed to explore the clinical-pathologic similarities and differences in the IBD-associated CRC (IBD-CRC) between patients in China and Canada. METHODS: Data of 78 patients with IBD-CRC retrospectively retrieved from two representative medical institutions in Beijing (China) and Calgary (Canada) over the same past 13 years, including 25 (22 UC-associated and three CD-associated) from Beijing group and 53 (32 UC-associated and 21 CD-associated) from Calgary group, were compared with regards to their clinical and pathologic characteristics. RESULTS: Several known features of IBD-CRC were seen in both groups, including long duration and large extent of colitis, active inflammation background, multifocal lesions, and advanced tumor-node-metastasis stage. Beijing group showed a significantly higher percentage of UC (88.0% vs. 60.4%, P = 0.018), younger age at diagnosis of CRC (48.6 ±â€Š12.8 years vs. 61.6 ±â€Š14.7 years, P < 0.001), lower ratio of mucinous adenocarcinoma (7.1% vs. 42.4%, P = 0.001) compared with Calgary group. None of the Beijing group had concurrent primary sclerosing cholangitis, while 5.7% of Calgary group did. Surveillance colonoscopy favored the detection rate of precancerous lesions (41.4% vs.17.0%, P = 0.002). CONCLUSIONS: As compared with patients from the Calgary group, the IBD-CRC patients in Beijing group were younger, less CD-associated and had less mucinous features, otherwise they were similar in many common features.

Pancreaticobiliary involvement in treated type 1 autoimmune pancreatitis: Imaging pattern and risk factors for disease relapse.

PURPOSE: To evaluate the imaging pattern of pancreaticobiliary lesions in patients with treated type 1AIP, to determine the incidence of disease relapse and malignancy, and to identify the risk factors. METHOD: The institutional review board approval was acquired. All patients gave written informed consent. From a prospective clinico-radiological database since 2012, consecutive patients with type 1 AIP who were treated and followed up (≥18 months) were identified. The presence/absence of pancreaticobiliary lesion(s) development during follow-up were assessed. The etiology was determined and the imaging pattern was compared to the initial attack. Risk factors were identified by univariate and multivariate analysis. RESULTS: Among 103 patients with treated type 1 AIP, 44 (42.7%) developed pancreaticobiliary lesions during follow up (median time interval to initial diagnosis: 17 months, range 3-62 months), mostly after steroid discontinuation (63.6%) or during maintenance therapy (29.5%). All lesions were disease relapse, which responded to steroid treatment. At relapse, pancreatic involvement was less frequent (81.8% vs 100%, p = 0.003), and the pancreas size was smaller (p < 0.01), whereas extra-pancreatic bile duct (ExPanBD) involvement was more severe and extensive (both p < 0.01). Multivariate analysis revealed ExPanBD involvement at initial diagnosis (hazard ratio 2.437, 95% CI 1.343-7.402, p = 0.002) and serum IgG4 response ratio at the induction phase (hazard ratio 0.357, 95% CI 0.055-0.804, p = 0.011) as significant independent predictors of relapse. CONCLUSIONS: In treated type 1 AIP, although imaging pattern may differ, pancreaticobiliary lesions are usually manifestations of disease relapse. ExPanBD involvement and poor serum response suggests high risk of relapse.

Clinical characteristics of ulcerative colitis-related colorectal cancer in Chinese patients.

OBJECTIVE: The prevalence of ulcerative colitis (UC) in China has been increasing, together with the incidence of ulcerative colitis-related colorectal cancer (UCRCC). In this study we aimed to investigate the clinical features of UCRCC in Chinese UC patients. METHODS: Clinical data of patients with UCRCC during January 2003 to December 2016 were retrieved from the Peking Union Medical College Hospital registration database and compared with that of sporadic colorectal cancer (CRC). RESULTS: During the study period, among the 10 110 patients with CRC 25 were found to have UCRCC, with an average age of 51 years and a median disease duration of 15 years. The cumulative prevalence of UCRCC was 0.25% in all CRC patients, with a significant increase from 2003-2007 to 2013-2016 (P < 0.01). Altogether 14 (56.0%) patients with UCRCC were men, and 5 (20.0%) had a family history of UC or cancer. Four (16.0%) patients had concurrent high-grade intraepithelial neoplasia with variable differentiated adenocarcinoma at other sites and 19 (59.4%) tumors were moderately or poorly differentiated. Twenty-three (92.0%) patients underwent surgery, of whom 10 (40%) were at stage III and IV according to the TNM staging system. Compared with the 10 085 patients with sporadic CRC, UCRCC patients were younger (P < 0.01) and were more likely to have descending colon involvement and multifocal lesions (P < 0.01). CONCLUSIONS: UCRCC patients have been increasing in recent years. Compared with patients with sporadic CRC, those with UCRCC are younger and are more likely to have descending colon involvement and multifocality.

Histological effects of unilateral spermatic cord torsion without removal of the ipsilateral necrotic testis on the contralateral testis in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of retarded removal of the unilateral necrotic testis after long-time (> 24 h) spermatic cord torsion on the contralateral testis in rats.</p><p><b>METHODS</b>Thirty-three male SD rats aged 21 -42 days were divided into a sham-operation group (n = 11), a torsion-reservation group (n = 12) and a torsion-orchiectomy group (n = 10). The rats of the sham-operation group received dartos pouch orchidopexy on the left testis, while those of the latter two groups underwent 720 degrees unilateral spermatic cord torsion on the left side. Ninety-six hours later, the rats of the torsion-reservation group received detorsion with the ipsilateral testis preserved, while those of the torsion-orchiectomy group underwent orchiectomy. Three months after operation, blood samples were obtained from the rats for measurement of serum testosterone and antisperm antibodies by ELISA, and meanwhile testes and epididymides were harvested for determination of the volumes of various structures and the diameter of seminiferous tubules with stereological methods.</p><p><b>RESULTS</b>There were no significant differences in the level of serum testosterone among the three groups. Anti-sperm antibody positive was found in only 1 animal in the torsion-reservation group. The Leydig cell nuclei in the contralateral testis appeared larger in the torsion groups than in the sham-operation group. Marked morphological changes were observed in 1, 3 and 0 of the animals in the sham-operation, torsion-reservation and torsion-orchiectomy group, respectively, mainly including atrophy of seminiferous tubules and reduced number of spermatogenic cells. The volume of the contralateral testis was increased by 19% and 21% in the torsion-reservation and torsion-orchiectomy group, respectively, in comparison with that in the sham-operation group (P < 0.05). No significant differences were observed in the volume of seminiferous tubules of the contralateral testis among the sham-operation, torsion-reservation and torsion-orchiectomy groups ([1.15 +/- 0.07], [1.30 +/- 0.04] and [1.35 +/- 0.05] cm3). The volume of the interstitial tissue was significantly increased in the latter two groups ([0.36 +/- 0.02 and 0.34 +/- 0.03] cm3) as compared with the former ([0.25 +/- 0.02] cm3) (P < 0.05). The diameters of the seminiferous tubules exhibited no significant differences among the three groups ([226.00 +/- 7.00], [223.00 +/- 6.00] and [221.00 +/- 3.0] microm).</p><p><b>CONCLUSION</b>Long-time unilateral spermatic cord torsion may result in compensatory hypertrophy of the contralateral testis, and orchiectomy does not significantly affect the histology of the contralateral testis and epididymis.</p>

Impact of ethane dimethane sulfonate on the histological structures of seminal vesicles in adult rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effect of ethane dimethane sulfonate (EDS) injection on the volumes of different histological structures in the seminal vesicles of adult rats.</p><p><b>METHODS</b>Twenty-seven male SD rats aged approximately 90 days were randomly divided into a control group (n = 14) and an EDS group (n = 13) to receive one intraperitoneal injection of normal saline and EDS (75 mg/kg bodyweight), respectively. At 7 and 12 days after treatment, the unilateral seminal vesicles were removed, methacrylate resin-embedded sections prepared and the total volumes of various structures in the seminal vesicles estimated using stereological methods.</p><p><b>RESULTS</b>EDS treatment almost completely destroyed the Leydig cells in the testis, resulting in a drastic testosterone deficiency. The volume of the seminal vesicle (including the coagulating gland attached to the vesicle) was decreased by 53% in the 7 d EDS group (n = 6) in comparison with the 7 d control group (n = 7) ([138.2 +/- 12.9] vs [64.9 +/- 3.6] mm3, P < 0.01), but showed no significant difference between the 7 d and the 12 d EDS (n = 7) groups ([64.9 +/- 3.6] vs [55.4 +/- 7.7] mm3, P > 0.05). The total volumes of the glandular lumen, glandular epithelium, smooth muscular layer and adventitia were decreased by 96.7, 80.3, 57.6 and 67.0%, respectively, in the 12 d EDS group as compared with the 12 d control group (n = 7).</p><p><b>CONCLUSION</b>EDS induces drastic testosterone deficiency in adult rats, and significantly reduces the total volumes of the seminal vesicle lumen, glandular epithelium, smooth muscular layer and adventitia.</p>

[Toxic megacolon complicated by ulcerative colitis in six patients: a case report and literature review].

OBJECTIVE: To summarize the clinical features of ulcerative colitis (UC) complicated by toxic megacolon for early diagnosis and proper treatment. METHODS: Six cases of toxic megacolon in the patients suffered from UC in Peking Union Medical College Hospital from 1983 to 2010 were analyzed, and related literature was searched and reviewed. RESULTS: The incidence of the toxic megacolon in the patients with UC in our center was 0.7%(6/824), which was lower than those reported in the literature. There were always risk factors triggering the disease. The prognosis of the patients was poor, even after medical care and surgery intervention. Evaluation of the patients and making right timing to perform the surgery would improve the prognosis of the patients in foreign literature. CONCLUSION: It's crucial to make early diagnosis of the toxic megacolon in the patients suffered from UC. The right choice and timing to perform urgent surgery or selective surgery may improve their prognosis.

Testosterone-induced spermatogenic impairment is associated with looser arrangement of spermatogenic cells in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine whether testosterone-induced intra-testicular testosterone withdrawal and therefore spermatogenic impairment is associated with looser arrangement of spermatogenic cells in rats.</p><p><b>METHODS</b>Adult male SD rats received intramuscular injection of testosterone undecanoate at 19 mg/(kg x 15 d) for 130 days, and then testicular tissue blocks were obtained for the preparation of methacrylate resin-embedded sections and observation of the changes in testicular histology.</p><p><b>RESULTS</b>Apart from such changes as impaired spermiogenesis and spermiation, apparently looser arrangement of spermatogenic cells was seen in 11.5% of the seminiferous tubule profiles, with radial cracks (empty spaces) running towards the tubule lumen being formed between lines, bundles or groups of spermatogenic cells (mainly spermatids and spermatocytes).</p><p><b>CONCLUSION</b>Looser arrangement of spermatogenic cells is one of the key histological changes resulting from intra-testicular testosterone withdrawal in rats.</p>

[The trends in clinical characteristics of colon cancer in last two decades].

OBJECTIVE: To explore the changing of clinical features of colon cancer within 20 years, in order to help early diagnosis and screening of colon cancer in China. METHODS: A total of 1233 cases of colon cancer in Peking Union Medical College Hospital during 1989 - 2008 were retrospectively studied. All patients were divided into two groups according to the date of onset (1989 - 1998 and 1999 - 2008), the demographic features, clinical manifestations, laboratory examination, colonoscopy characteristics and pathological stage were analyzed. RESULTS: Comparing with 1989 - 1998, in recently 10 years, the morbidity of colon cancer increased, more female and old patients appeared; hematochezia significant less (51.8% vs 31.7%, P < 0.05); abdominal mass and following ileus also decreased (30.2% vs 13.6%, P < 0.05); patients with low of Hb decreased, the positive of stool occult blood increased from 43.6% to 61.2% (P < 0.05), According with the more patients who detected serum CEA, the positive rate significant increased (32.4% vs 57.9%, P < 0.05). Colonoscopy became the mainly method for diagnosis, more and more early stage lesion and polyps were detected. The location of tumor from ascending colon in 1989 - 1998 (44.6%) shift to sigmoid colon (38.7%) and descending colon (22.7%) up to now. Operation was the first choice of treatment, the early stage (Duke A) patients significant increased (9.3% vs 23.8%, P < 0.05). CONCLUSIONS: In the recently 10 years, the morbidity of colon cancer obviously increased, the age was become elder and female patients were increased. The clinical manifestation became more nonspecific. According with the improvement of stool occult blood, serum CEA and colonoscopy detective method and wild spread using, more and more early stage patients were diagnosed. The location of tumor shift from right side to left side, and coincidence with west countries gradually.

[Pleuropulmonary blastoma in an adult women with pleurorrhea as the major clinical manifestation: report of a case].

OBJECTIVE: Pleuropulmonary blastoma (PPB) is a rare dysontogenetic neoplasm in children and has been recognized as a distinct clinicopathological entity different from the ordinary pulmonary blastoma of adulthood. We report a very rare adult case of PPB. A 43-year-old female patient presented with massive pleural infusion, and a misdiagnosis of pleural tuberculosis (TB) was made on the basis of ultrasound scan and CT images, for which therapy with TB drugs was administered for 3 weeks. Subsequent operation and pathologic examination of the mass revealed a multicystic neoplasm consisting of malignant mesenchymal cells immunohistochemically positive for vimentin and actin. Local recurrence occurred in the left pleural 20 months after the surgical tumor resection and 4 cycles of adjuvant chemotherapy.

Effect of vasectomy via inguinal canal on spermatogenesis in rabbits / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To determine whether vasectomy away from the epididymal tail (via the inguinal canal) in rabbits can reduce the early postoperative effects on spermatogenesis.</p><p><b>METHODS</b>Twenty-nine normal male Japanese white rabbits (aged 4-6 months) were subjected to unilateral close-ended (conventional) or open-ended (the cut end of the juxta-epididymal vas deferens not ligated) vasectomy via the inguinal canal. Ten days and 3 months after operation, testes, epididymides and vasa deferentia were removed and methacrylate resin-embedded sections prepared. The histology of the testis, epididymis and vas deferens was examined under light microscope, and the volume and diameter of the seminiferous tubules were quantitatively studied using stereological methods.</p><p><b>RESULTS</b>Neither of the methods of vasectomy led to apparent damage to spermatogenesis on the vasectomized side in comparison with the contralateral sham-operated side, but the juxta-epididymal vas deferens on the vasectomized side was highly distended and contained numerous sperm 3 months after operation.</p><p><b>CONCLUSION</b>Vasectomy away from the cauda epididymis has no significant early postoperative effects on spermatogenesis in rabbits.</p>

[Are leukemic patient bone marrow mesenchymal stem cells malignant?].

The study was aimed to explore whether there are leukemic characteristics in the bone marrow mesenchymal stem cells (BMMSC) from leukemic patients as compared with normal controls. The mesenchymal stem cells from bone marrow of normal volunteers and patients with APL and CML were isolated, then cultured and proliferated in vitro. The morphology, growth curve and cell surface markers of two different sources mesenchymal stem cells were investigated for detecting whether the bone marrow mesenchymal stem cells derived from leukemia patients have the specific abnormal fusion gene of leukemia cells through fluorescent in situ hybridization. The results indicated that there was no significant difference between the mesenchymal stem cells derived from different subjects, the bone marrow mesenchymal stem cells derived from leukemia patients did not have the clonal malignant fusion gene as seen in the leukemia cells. Taken altogether, mesenchymal stem cells derived from leukemia patients had no biological differences as compared with those from normal volunteers, and no malignant clonal abnormality was found. It is concluded that mesenchymal stem cells derived from leukemia patients as an alternative vehicle may be used for assistant of autologous hematopoietic stem cell transplantation or cell therapy and gene therapy.

Ultrasonographic changes in the epididymis after long-term vasectomy / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the ultrasonographic changes in the epididymis after long-term vasectomy.</p><p><b>METHODS</b>Sixty-four patients with a history of vasectomy for more than 10 years (vasectomy group) and another 60 without vasectomy (control group) were included in the study. The patients were referred to scrotal ultrasonography for epididymal indications. The shape, thickness and internal echoes of the head, body and tail of the epididymis were observed with high frequency ultrasonography (HFU), and the blood flow changes were observed with color Doppler flow imaging (CDFI) or color Doppler power imaging (CDPI).</p><p><b>RESULTS</b>Significantly higher rates were found in the vasectomy group than in the control: thickened body (64.1% vs 15.0%) and tail (78.1% vs 51.7%) of the epididymis, thickened head, body and tail (42.2% vs 8.3%) of the epididymis, and epididymal tubular ectasia (54.7% vs 8.3%). However, increased blood flow in the epididymis was seen at a significantly lower rate in the vasectomy group than in the control (15.6% vs 61.7%).</p><p><b>CONCLUSION</b>The ultrasonographic changes in the epididymis after long-term vasectomy were mainly epididymis thickening and epididymal tubular ectasia, mostly with no or diminished blood flow in the epididymis.</p>

Histological changes of the testis and epididymis in adult rats as a result of Leydig cell destruction after ethane dimethane sulfonate treatment: a morphometric study / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion.</p><p><b>METHODS</b>Fourteen adult Sprague-Dawley rats were injected intraperitoneally with ethane dimethane sulfonate (EDS, 75 mg/kg) and the same number of animals were injected with normal saline as a control. At days 7 and 12 (after treatment), respectively, half of the animals from each group were killed. The testes and epididymides were removed and tissue blocks embedded in methacrylate resin. The cell number per testis was estimated using the stereological optical disector and some other parameters were obtained using other morphometric methods.</p><p><b>RESULTS</b>The EDS treatment resulted in an almost complete elimination of Leydig cells but had no effect on the numbers of Sertoli cells per testis. At day 7 after EDS treatment, many elongated spermatids were retained in the seminiferous epithelium and many round spermatids could be seen in the epididymal ducts. At day 12, a looser arrangement of spermatids and spermatocytes became evident, with apparent narrow empty spaces being formed between germ cells in an approximately radial direction towards the tubule lumen; the numbers (per testis) of non-type B spermatogonia and spermatocytes were similar to controls, whereas that of type B spermatogonia increased by 59%, and that of early round, elongating and late elongated spermatids decreased by 37%, 72% and 52%, respectively.</p><p><b>CONCLUSION</b>The primary spermatogenic lesions following EDS administration were (i) spermiation failure and (ii) detachment of spermatids and spermatocytes associated with impairment in spermiogenesis and meiosis.</p>

Effect of intra-testicular testosterone withdrawal on the expression of alpha-catenin in the adult rat testis / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the expression of alpha-catenin in the rat testis after intra-testicular testosterone withdrawal induced by injection of testosterone undecanoate (TU).</p><p><b>METHODS</b>Ten adult male SD rats received vehicle (n = 5 ) or TU (19 mg/kg every 15 days, n = 5) for 130 days. Paraffin-embedded testicular sections were used for immunohistochemistry against a polyclonal anti-alpha-catenin antibody.</p><p><b>RESULTS</b>In the control, alpha-catenin was expressed in the acrosome of spermatids and the cytoplasm of Leydig cells and peritubular myoid cells. In the TU-treated rat testis, Leydig cells were atrophied and the expression of alpha-catenin was markedly decreased or absent, but there was no evident change in the immunostaining of spermatids or myoid cells.</p><p><b>CONCLUSION</b>Intra-testicular testosterone withdrawal-induced looser arrangement or sloughing of spermatogenic cells is not related to the adhesion molecule alpha-catenin. Alpha-catenin may be used as a cell identification marker for Leydig cells.</p>

[Analysis of sequence-tagged site in bcr and abl genes by DNA pooling and dHPLC].

To investigate the relationship between the single nucleotide polymorphism (SNPs) of the bcr and abl gene and chronic myelogeous leukemia (CML), the 9 sequence-tagged sites (STS) in bcr and abl gene were screened by DNA pooling and denaturing high performance liquid chromatography (dHPLC), and the results were varified by sequencing. The results showed that the polymorphism sites were detected in 4 out of the 9 STS fragments and there were 3 bases different from the reference sequence found in 3 fragments. In conclusion, the novel SNP in U07000 fragment shows significantly different frequencies between CML and controled people.

[Recombinant adeno-associated virus 2-mediated green fluorescent protein expression in bone marrow mesenchymal stem cells derived from acute myelogenous leukemia patients].

OBJECTIVE: To explore the possibility of using autologous bone marrow mesenchymal stem cells (BMSC) as a vehicle to deliver recombinant adeno-associated virus 2-mediated enhanced green fluorescent protein (rAAV-2-eGFP) in vitro, therefore to find an alternative solution for gene therapy of hematological malignancy. METHODS: BMSCs isolated from the bone marrow of patients with acute myelogenous leukemia (AML) at the onset of disease were infected by rAAV-2-eGFP at different multiplicity of infection (MOI=10(2), 10(3), 10(4), 10(5), 10(6), and 10(7), respectively). Phase-contrast fluorescent microscope and flow cytometry were employed to evaluate the expression of enhanced green fluorescent protein (eGFP). RESULTS: Ten to fourteen days after the transfection, eGFP expression began to be detected and the transfection efficiency ranged between 0.3% to 2%, which failed to be increased with the increase of MOI. The transduced eGFP could maintain a long-term stable expression in vitro in the 61 days of observation, and from 12 to 33 days after transfection, eGFP percentage underwent a decrease from the initial 1.16% to 0.5%-0.6% and maintained this expression level till 61 days after transfection. CONCLUSION: rAAV can be used with BMSCs for in vitro gene therapy, but the poor transfection efficiency of these cells remains a significant obstacle for its further application.

[Polymorphism analysis of 5' promotor region of BCR gene].

BACKGROUND & OBJECTIVE: BCR-ABL fusion gene is regarded as the molecular hallmark of chronic myelogenous leukemia (CML), and its expression is controlled by the BCR gene promoter. This study was designed to investigate the polymorphism of the promoter region of BCR gene, and its possible correlation with the disease. METHODS: A 1.13 kb fragment of BCR gene 5' promotor region was amplified and sequenced from 30 CML patients and 19 controls. Transcription factor binding sites and repeat sequences in this region were analyzed using softwares and online tools. RESULTS: Four novel single nucleotide polymorphisms (SNPs) and 3 bases different from the reference sequence were detected in the region studied. Among these 2 novel SNPs and 1 different base were located in or near several bases of binding sites. The gene frequencies of the novel SNPs had no significant difference between CML and control people. CONCLUSION: Sequence polymorphisms were found in the 5' promotor region of BCR gene, most of them being SNPs. No relativity can be validated between the SNPs and the disease. But it appears that some SNPs might have the probability of bringing influence to the transcription and expression of the gene.