[The Origin and Academic Characteristics of Xujiang School].

Xujiang School of acupuncture and moxibustion has a long history with distinctive academic characteristics and regional influence. Xujiang School, originated from Xi Hong in Song Dynasty, is the oldest acupuncture and moxibustion school recorded in Chinese history. Later, it was passed down from family to family for more than ten generations. The tenth generation Xi Xinqing passed it on to Chen Honggang and gradually evolved into a school of acupuncture and moxibustion with regional characteristics and a certain national influence. In terms of academic characteristics, doctors in Xujiang School kept innovating based on the Classics.Its acupuncture and moxibustion academic ideas including reinforcement and reduction , point selection and searching for the primary cause of disease in treatment have had an important impact on contemporary acupuncture in clinic.

Structural and functional comparisons of the environmental microbiota of pond and tank environments at different locations for the commercial aquaculture of American shad.

The environmental microbiota plays a significant role in the growth and development of aquatic life. In recent years, American shad has become an important economic aquaculture species in China. However, information on the correlation between the growth of American shad and the aquaculture environment is limited. Through 16S rDNA-based sequencing, the microbiota communities in ponds at different locations (Jiangyin and Yancheng in Jiangsu, China) were investigated. The results showed that the richness and diversity of the microbiota in the pond were greater than those in the tank at the same location. At the phylum level, Proteobacteria and Firmicutes were more abundant in the samples from Jiangyin than in those from Yancheng. In addition, the ratio of Firmicutes to Bacteroidetes was larger in the JYT samples than in the YCT samples, which implied that the American shad cultured in the Jiangyin environment tended to be fatter than those cultured in Yancheng. The body weight data also confirmed this finding. Moreover, the proportions of functional annotations in the samples from the Jiangyin and Yancheng environments were similar, but there were differences between the overall levels. Our results highlighted the correlations between the environmental microbiome and the growth tendency of American shad.

[Selection and effects of flap/myocutaneous flap repair methods for the defect after perineum tumor resection].

Objective: To explore the selection and effects of flap/myocutaneous flap repair methods for the defect after perineum tumor resection. Methods: From January 2011 to February 2017, 31 patients with vulvar tumor who were admitted to Hunan Cancer Hospital underwent repair of wound after tumor resection with various flaps/myocutaneous flaps. The patients were composed of 5 males and 26 females, aged 39-76 years, with 27 vulvar cancer and 4 Paget's disease in primary diseases. The size of defects after vulvar tumor radical resection ranged from 8.0 cm×4.5 cm to 27.5 cm×24.0 cm. According to the theory of perforasome, the defects were repaired by the external pudendal artery perforator flap, deep inferior epigastric artery perforator flap, rectus abdominis myocutaneous flap, anterolateral thigh flap, internal pudendal artery perforator flap, gracilis myocutaneous flap, and profunda artery perforator flap based on the specific size and location of perineum and groin where the defect was located. According to the blood supply zone of flap, totally 17 local translocation flaps, 18 axial flaps/myocutaneous flaps, and 7 V-Y advancement flaps were resected, with an area of 7.0 cm×4.0 cm to 21.0 cm×13.0 cm. All the flaps/myocutaneous flaps were transferred in pedicled fashion, and the donor sites were closed without tension. The number of flaps/myocutaneous flaps, wound closure, flaps/myocutaneous flaps survival, and follow-up were observed and recorded. Results: Altogether 42 flaps/myocutaneous flaps were harvested in 31 patients. Two flaps/myocutaneous flaps were used in 11 cases for large circular defect repair. All the defects achieved tension-free primary closure. The blood supply of 32 flaps/myocutaneous flaps was good, while insufficient blood supply was noted in the other 10 flaps/myocutaneous flaps. Seventeen flaps/myocutaneous flaps survived smoothly. Wound dehiscence occurred in 5 flaps/myocutaneous flaps 8 to 14 days postoperatively, which was healed with dressing change. Temporary congestion was noted in 7 flaps/myocutaneous flaps 2 to 5 days postoperatively, which recovered without special treatment. Three flaps/myocutaneous flaps had infection 7 to 15 days postoperatively, two of which recovered after dressing change, while the other one had partial necrosis and received debridement and direct closure. Two flaps/myocutaneous flaps were totally necrotic 8 to 15 days postoperatively, which were repaired with pedicled rectus abdominis myocutaneous flap after debridement. Seven flaps/myocutaneous flaps had partial necrosis 7 to 20 days postoperatively and were healed after dressing change. Twenty-four patients were followed up for 9-38 months. The color of flaps/myocutaneous flaps was similar to that of the surrounding skin, the shape of vulva was natural, the movement of hip joint was not limited, the function of micturition and defecation was not affected, and tumor recurrence was noted in 3 patients. Conclusions: For the complicated large defect after perineum tumor resection, the flexible application of different forms of flaps/myocutaneous flaps to repair according to different areas regains the appearance and function. However, there are many complications, so it is necessary to further strengthen the postoperative care.

Granny Smith apple procyanidin extract upregulates tight junction protein expression and modulates oxidative stress and inflammation in lipopolysaccharide-induced Caco-2 cells.

The present work is undertaken to characterize a Granny Smith apple procyanidin extract (AE) and investigate the beneficial effect of the AE in the intestine in vitro. Each AE was characterized via LC-ESI-MS. Caco-2 cells were used to study the preventive actions of the AE against the downregulation of tight junction protein expression, oxidative stress and inflammation induced by lipopolysaccharides (LPS). Phenolic compounds present in the AE, including chlorogenic acid, catechin, epicatechin, proanthocyanidin dimers, and proanthocyanidin trimers, were characterized. The expression of the tight junction protein, including occludin and zona occludens (ZO)-1, increased significantly in LPS + AE treated Caco-2 cells, compared to LPS induced Caco-2 cells. Proanthocyanidin dimers had the most potent effect on increasing tight junction protein expression. The addition of LPS to Caco-2 cells induced oxidative stress and inflammation. However, incubation with proanthocyanidin dimers prevented LPS-mediated oxidative stress, including the increase of SOD, HO-1, CAT, and GSH-Px mRNA expression, and counteracted LPS-mediated inflammation as evidenced by the down-regulation of inflammatory markers (NF-κß, IL-6, and TNF-α mRNA expression). Our findings provide evidence that AE could upregulate tight junction protein expression, probably acting via the reduction of oxidative stress and inflammation.

Prediction of fecal nitrogen and phosphorus excretion for Chinese Holstein lactating dairy cows.

Mathematical models for predicting nitrogen and phosphorus excretion play a key role in manure application and environment monitoring. An analysis for prediction of fecal nitrogen (FN, g/d) and fecal phosphorus (FP, g/d) excretion for Chinese Holstein lactating dairy cows was conducted using a data set from 15 dairy farms in northern China. The whole independent-variable data set, obtained with questionnaires, consisted of 110 sets of average diet nutrient compositions, including DMI (kg/d), CP content (% DM), OM intake (OMI, kg/d), nitrogen intake (NI, g/d), and phosphorus intake (PI, g/d), and animal characteristics, including average days in milk (DIM, d), average milk yield (MY, kg/d), and average BW (kg). In addition, 110 fecal samples in total were collected to analyze FN and FP excretions, which were considered dependent variables of prediction equations. Correlations between diet and animal variables were examined, and several variable subpools were derived that were used to develop equations to predict FN and FP excretions by stepwise regression analysis. The results showed that among all variables, OMI was the best predictor for FN excretion (root-mean-square prediction error [RMSPE] = 9.58%, = 0.70), followed by NI (RMSPE = 10.19%, = 0.67). However, when both DMI and CP were used as independent variables, the equation showed more accurate prediction for FN excretion (RMSPE = 8.55%, = 0.77) in comparison with univariate prediction equations. Simultaneously, PI was the best predictor of FP excretion (RMSPE = 10.28%, = 0.67). Evaluation results using 3-fold cross validation and comparison with extant equations indicated that the proposed equations were accurate with low prediction errors, which could be recommended for use to estimate FN and FP excretions from Chinese Holstein lactating dairy cows.

Induction of trap formation in nematode-trapping fungi by bacteria-released ammonia.

A total of 11 bacterial strains were assayed for bacteria-induced trap formation in the nematode-trapping fungus Arthrobotrys oligospora YMF1·01883 with two-compartmented Petri dish. These strains were identified on the basis of their 16S rRNA gene sequences. Volatile organic compounds (VOCs) of eight isolates were extracted using solid-phase micro-extraction (SPME) and their structures were identified based on gas chromatography-mass spectrometry (GC-MS). At the same time, all isolates were used for quantitative measurement of ammonia by the indophenol blue method. The effects of pure commercial compounds on inducement of trap formation in A. oligospora were tested. Taken together, results demonstrated that the predominant bacterial volatile compound inducing trap formation was ammonia. Meanwhile, ammonia also played a role in other nematode-trapping fungi, including Arthrobotrys guizhouensis YMF1·00014, producing adhesive nets; Dactylellina phymatopaga YMF1·01474, producing adhesive knobs; Dactylellina cionopaga YMF1·01472, producing adhesive columns and Drechslerella brochopaga YMF1·01829, producing constricting rings.

Effect of DDAH/ADMA/NOS regulation pathway on cavernae corporum cavernosorum rat penis of different age.

The effect of DDAH/ADMA/NOS pathway in penile tissue of rats of different age was investigated to better understand the mechanism of age-related erectile dysfunction (ED). The Sprague Dawley male rats were assigned as the young group (3 month old, n = 10) and the old group (18 month old, n = 10) respectively. Intracavernous pressure (ICP) was measured before and after papaverine intracavernous injection. Pathology structure of penile tissue was evaluated under transmission electron microscope. The expression amounts of asymmetric dimethylarginine (ADMA) and cyclic guanosine monophosphate (cGMP) in penile tissue were detected by ELISA; the expression levels of isoform-specific DDAH and NOS were assessed via Western blot. Compared with the young group, the ICP in the old group rat decreased significantly (33.46 ± 5.37 versus 39.71 ± 3.67 mmHg, P = 0.02) after papaverine injection. Diffused fibrosis and impairment of endothelial cell were observed in corpus cavernosum in the old group rats. Higher level of ADMA (10.83 ± 0.96 versus 7.51 ± 1.39 µmol per gpro, P = 3.14 × 10(-4) ) and lower level of cGMP (29.42 ± 3.84 versus 47.09 ± 6.07 nmol per gpro, P = 1.57 × 10(-6) ) were detected in penile tissue of the old group compared with those of the young group. Expression of DDAH1, DDAH2, endothelial NOS (eNOS) and neuronal NOS(nNOS) all decreased significantly in penile tissue of the old group rat. The DDAH/ADMA/NOS regulation pathway changes dramatically accompanying with lower ICP in old group rat compared with those of the young group. Such findings in rats are suggestive in understanding the mechanism of age-related ED in humans.

Cloning and heterologous expression of UDP-glycosyltransferase genes from Bacillus subtilis and its application in the glycosylation of ginsenoside Rh1.

UNLABELLED: Bacillus subtilis CCTCC AB 2012913 can transform ginsenoside Rh1 to 3-O-ß-D-glucopyranosyl-6-O-ß-D-glucopyranosyl-20(S)-protopanaxatriol. Based on its genome sequence, strain B. subtilis 168 contains three UDP-glycosyltransferase genes. Here, we cloned the three UDP-glycosyltransferase genes (ydhE1, yojK1 and yjiC1) from B. subtilis CCTCC AB 2012913 and expressed in Escherichia coli BL21 (DE3) with His-tag. The crude enzyme extracts were assayed, respectively, for their activities to transform ginsenoside Rh1. Extracts containing enzymes YojK1 and YjiC1 could use ginsenoside Rh1 as a substrate to produce 3-O-ß-D-glucopyranosyl-6-O-ß-D-glucopyranosyl-20(S)-protopanaxatriol, which had an additional glucopyranosyl linked with C-3 over the substrate. Enzyme YjiC1 was purified by affinity chromatography on Ni-NTA His Binding resin. The molecular mass of purified YjiC1 was c. 47 kDa as determined by SDS-PAGE. This is the first report of an in vitro biotransformation of ginsenoside Rh1 to 3-O-ß-D-glucopyranosyl-6-O-ß-D-glucopyranosyl-20(S)-protopanaxatriol using the recombinant UDP-glycosyltransferase. SIGNIFICANCE AND IMPACT OF THE STUDY: The Chinese traditional medicinal plant Panax is reported to have multiple health benefits. Its main active ingredient is saponin, and different saponins have different activity spectrum. In the study, three UDP-glycosyltransferase genes, ydhE1, yojK1 and yjiC1, were cloned from Bacillus subtilis CCTCC AB2012913 and the three genes were expressed in Escherichia coli BL21 (DE3). The enzyme YjiC1 was purified and converted ginsenoside Rh1 to 3-O-ß-D-glucopyranosyl-6-O-ß-D-glucopyranosyl-20(S)-protopanaxatriol in vitro. The compound is the first saponin possessing ß-glucopyranosyl at both C-3 and C-6 sites. We showed that the in vitro biotransformation was effective, and the reaction condition was easy to control. Our research suggests that a diversity of saponins could be generated through efficient and directed enzymatic biotransformation.

Management of Meloidogyne incognita on tomato with endophytic bacteria and fresh residue of Wasabia japonica.

AIMS: To characterize the nematicidal endophytic bacteria (NEB) of Wasabia japonica (wasabi) and evaluated the control efficacies of promising NEB as well as fresh wasabi residue (FWR) against Meloidogyne incognita on tomato. METHODS AND RESULTS: By in vitro bioassay, 53 NEB strains showing nematicidal efficacies of >50% against J2 of M. incognita were isolated from wasabi. Basing on 16S rRNA gene sequences, these NEB were identified into 18 species of 11 genera. In greenhouse, incorporation of selected NEB culture or FWR into potted soil significantly reduced infection of M. incognita on tomato. Treating tomatoes with either FWR or NEB of Raoultella terrigena RN16 and Pseudomonas reinekei SN21 in the field yielded excellent control efficacies against M. incognita, especially the combinations of FWR with either R. terrigena RN16 or Ps. reinekei SN21 at doses of 50 g plus 100 ml per plant or more. CONCLUSIONS: The results established that R. terrigena RN16 and Ps. reinekei SN21 applied separately or combined with FWR have the potential to provide bioprotection agents against M. incognita. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides novel way for disease management using combination of endophyte and host residue.

Clinical analysis of the characterization of magnetic resonance imaging in 102 cases of refractory haematospermia.

To analyze the pathogenesis of persistent and refractory haematospermia and to evaluate the aetiological diagnostic value of magnetic resonance imaging (MRI) for this type of haematospermia. Clinical data from 102 patients with persistent and refractory haematospermia was retrospectively analysed. Data collected included history, symptoms, as well as ultrasound and MRI of the morphological features of the bilateral seminal vesicles (SV) and ejaculatory duct (ED) areas. Criteria for inclusion were haematospermia symptoms that occurred more than six times, that lasted more than 6 months, and that did not improve after >1 month of conservative treatment. Patients underwent seminal vesiculoscopy with a post-surgery follow-up of 3-48 months [average (18.1 ± 10.3) months]. Of the 102 patients that underwent MRI examination, data from 88 patients (86.3%) showed typical and characteristic changes in the ED area, including the signal intensity changes in 60 (58.8%), SV volume changes in 32(31.4%), the formation of cysts such as prostatic utricular cysts in 27 (26.5%), Müllerian cysts in 4 (3.9%), ED cysts in 5 (4.9%) and a SV cyst in 1(1.0%). The MRI findings were confirmed by seminal vesiculoscopy and all patients received appropriate treatment. In 14 patients (13.7%), no obvious abnormal changes were observed with MRIs, however, these patients were diagnosed and successfully managed using seminal vesiculoscopy. Some degrees of ED obstruction was usually found during surgery. The symptoms of haematospermia disappeared 1-2 months after surgery in all patients. Two patients had a recurrence of haematospermia, underwent the same treatment, and recovered during the follow-up period. The aetiology of the most cases of the refractory haematospermia can be distinguished using the three-dimensional MRI. Typical abnormalities observed on MR images are signal intensity, SV volume changes and cyst formation. MRI has significant etiological diagnostic value and provides reliable information for the subsequent treatment of patients with persistent and refractory haematospermia.

Heme oxygenase-1 expression and its significance for acute rejection following kidney transplantation in rats.

OBJECTIVE: To analyze rejection and antiapoptotic effects of heme oxygenase-1 (HO-1) in kidney transplantations, to investigate the protective effects of endogenous HO-1 induced by hemin on acute rat kidney allograft rejection. METHODS: We selected 27 Brown-Norway rats and 27 male Lewis rats as donors and recipients, respectively, randomly dividing them into three groups: kidney transplantation alone, hemin treatment group, and cyclosporine (CsA) group (n = 18). Six recipient rats were harvested on the first, fifth, or seventh days after operation among each group to examine histopathologic changes in renal tissue, HO-1 protein expression, and acute rejection as well as to measure serum creatinine values. RESULTS: HO-1 expression in both the kidney transplantation model group and the hemin-induced groups were higher compared with the CsA group (P < .05-.01). The expression increased with the aggravation of rejection; the expression in the CsA group also increased after transplantation but was obviously lower than that of the hemin-induced group (P < .01). The rejection process was relatively mild as indenset by histopathologic examination. The serum creatinine levels among the hemin-induced group were lower compared to the kidney transplantation control group (P < .05), but higher compared to the CsA group (P < .05). CONCLUSION: HO-1 provided protection of allografts against rejection in rats, but such effects were poorer than those achieved using potent immunosuppressive agents like CsA.

Urological complications in 1,223 kidney transplantations.

INTRODUCTION: Urological complications after kidney transplantation may lead to graft loss. In this study, we retrospectively reviewed urological complications in 1,223 kidney transplantations that were performed at our institution. MATERIALS AND METHODS: The occurrence of urological complications such as urine leakage, ureteral obstruction and vesicoureteral reflux (VUR) according to the different way of urinary tract reconstruction, ureteroneocystostomy (U-C) and ureteroureterostomy (U-U), was studied. RESULTS: Urological complications were encountered in 92 (7.5%) cases, including urine leakage (n = 43, 3.5%), ureteral obstruction (n = 35, 2.9%) and VUR (n = 14, 1.1%). 75 cases (7.9%) were in the U-C group and 17 cases (6.2%) in the U-U group. 91 recipients were successfully treated, and 1 patient lost the graft due to kidney pelvis and ureteral necrosis. There was no recipient loss due to these complications. For recipients with urological complications, the 1- and 3-year survival rates were 90 and 88% for recipients and 87 and 82% for grafts, respectively. CONCLUSIONS: After U-U, the same number of overall incidences of urological complications is observed as after U-C; however, a decrease in the number of incidences of urine leakage is apparent. Therefore, U-U is a good first option with a greater success rate of resolving ureteral stenosis with endourology and no risk of VUR.

Treatment of urinary fistula after kidney transplantation.

Urinary fistula is a common complication after kidney transplantation and may lead to graft loss and patient death. Its current incidence ranges from 1.2% to 8.9%. From December 1993 to April 2007, 1223 kidney transplant procedures were performed by our kidney transplantation team. In 948 recipients (group 1), we performed an extravesical ureteroneocystostomy, and in 275 recipients (group 2), a terminoterminal ureteroureterostomy (UU). We observed urinary fistulas in 43 patients (3.5%), with mean onset at 6 days (range, 3-20 days) posttransplantation. Urinary fistula was significantly more common in group 1 compared with group 2 (4.1% and 1.5%, respectively; P < .05). The distal ureteral necrosis was the major frequent cause of urinary fistula (n = 34; 76.7%), which required either a second ureteroneocystostomy or UU using the native ureter. Of these 21 fistulas, including 10 recurrent fistulaes, were successfully treated with pedicled omentum covering the anastomotic stoma. Conservative treatment with a stent and Foley catheter drainage for 1 to 2 weeks was successful in 8 patients. All patients with a urinary fistula regained normal graft function except 1 in whom transplant nephrectomy was necessary because of pelvic and ureteral necrosis. There was no recipient loss secondary to urinary fistula. In conclusion, UU can decrease the incidence of urinary fistula after kidney transplantation. Most urinary fistulas require surgical management; and pedicled omentum is useful to repair the fistula.

Cloning and phylogenetic analysis of the chitinase gene from the facultative pathogen Paecilomyces lilacinus.

AIMS: To PCR-amplify the full-length genomic-encoding sequence for one chitinase from the facultative fungal pathogen Paecilomyces lilacinus, analyse the DNA and deduced amino acid sequences and compare the amino acid sequence with chitinases reported from mycopathogens, entomopathogens and nematopathogens. METHODS AND RESULTS: The encoding gene (designated as PLC) was isolated using the degenerate PCR primers and the DNA-Walking method. The gene is 1458 bp in length and contains three putative introns. A number of sequence motifs that might play a role in its regulation and function had also been found. Alignment of the translation product (designated as Plc, molecular mass of 45.783 kDa and pI of 5.65) with homologous sequences from other species showed that Plc belongs to Class V chitinase within the glycosyl hydrolase family 18. The phylogenetic and molecular evolutionary analysis using mega (Molecular Evolutionary Genetics Analysis) indicated that these chitinases from mycopathogens, entomopathogens and nematopathogens, the majority of which belong to glycosyl hydrolase family 18, were clustered into two well-supported subgroups corresponding to ascomycetes fungal and nonfungal chitinases (bacteria, baculoviruses). CONCLUSIONS: Our study showed that chitinases from mycoparasitic, entomopathogenic and nematophagous fungi are closely related to each other and reaffirmed the hypothesis that baculovirus chitinase is most likely to be of a bacterial origin - acquired by gene transfer. Bacterial and baculoviral chitinases in our study are potential pathogenicity factors; however, we still cannot ascribe any specific function to those chitinases from the fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report describing the chitinase gene and its translation product from Paecilomyces lilacinus, which constitutes the largest number of formulated biological nematicides reported so far, this is also the first study to analyse and resolve the phylogenetic and molecular evolutionary relationships among the chitinases produced by mycopathogens, entomopathogens and nematopathogens.

Proteases from Bacillus: a new insight into the mechanism of action for rhizobacterial suppression of nematode populations.

AIMS: The aim of this study was to investigate the role of proteases in Bacillus spp. of rhizobacteria in suppressing nematode populations and to understand their mechanism of action. METHODS AND RESULTS: Rhizobacteria with nematicidal activity were isolated from soil samples of five root knot nematode-infested farms. Among these strains, nematotoxicities of Bacillus strains were intensively analysed. Further assays of nematicidal toxins from Bacillus sp. strain RH219 indicated an extracellular cuticle-degrading protease Apr219 was an important pathogenic factor. The Apr219 shared high similarity with previously reported cuticle-degrading proteases from Brevibacillus laterosporus strain G4 and Bacillus sp. B16 (Bacillus nematocida). The cuticle-degrading protease genes were also amplified from four other nematicidal Bacillus strains isolated from the rhizosphere. In addition to Apr219, a neutral protease Npr219 from Bacillus sp. RH219 was also investigated for activity against nematodes. CONCLUSIONS: The wide distribution of cuticle-degrading proteases in Bacillus strains with nematicidal activity suggested that these enzymes likely play an important role in bacteria-nematode-plant-environment interactions and that they may serve as important nematicidal factors in balancing nematode populations in the soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Increased understanding of the mechanism of action of Bacillus spp. against nematodes could potentially enhance the value of these species as effective nematicidal agents and develop new biological control strategies.

Purification and characterization of an extracellular serine protease from the nematode-trapping fungus Dactylella shizishanna.

AIMS: To evaluate the production of an extracellular serine protease by Dactylella shizishanna and its potential as a pathogenesis factor. METHODS AND RESULTS: An extracellular alkaline serine protease (Ds1) was purified and characterized from the nematode-trapping fungus D. shizishanna using cation-exchange chromatography and hydrophobic interaction chromatography. The molecular mass of the protease was approximately 35 kDa estimated by SDS-PAGE. The optimum activity of Ds1 was at pH 10 and 55 degrees C (over 30 min). The purified protease could degrade purified cuticle of Penagrellus redivivus and a broad range of protein substrates. The purified protease was highly sensitive to phenylmethyl sulfonyl fluoride (PMSF) (0.1 mmol l(-1)), indicating it belonged to the serine protease family. The N-terminal amino acid residues of Ds1 are AEQTDSTWGL and showed a high homology with Aozl and PII, two serine proteases purified from the nematode-trapping fungus Arthrobotrys oligospora. CONCLUSIONS: Nematicidal activity of D. shizishanna was partly related to its ability to produce extracellular serine protease. SIGNIFICANCE AND IMPACT OF THE STUDY: In this report, we purified a new serine protease from D. shizishanna and provided a good foundation for future research on infection mechanism.

Expression and initial promoter characterization of PCAN1 in retinal tissue and prostate cell lines.

Prostate cancer is the most frequently diagnosed neoplasia in men and one of the leading causes of cancer-related deaths in men over 60. In an effort to understand the molecular events leading to prostate cancer, we have identified PCAN1 (prostate cancer gene 1) (also known as GDEP), a gene that is highly expressed in prostate epithelial tissue and frequently mutated in prostate tumors. Here we demonstrate its expression in neural retina, and retinoblastoma cell culture but not retinal pigment epithelial cell culture. We further characterize PCAN1 expression in the prostate cell lines RWPE1, RWPE2, and LnCAP FGC. We demonstrate an increase in expression when the cells are grown in the presence of Matrigel, an artificial extracellular basement membrane. Expression was time dependent, with expression observed on d 6 and little or no expression on d 12. Testosterone was not found to increase PCAN1 expression in this culture system. In addition, normal prostate epithelial cells co-cultured with normal prostate stromal cells did not exhibit PCAN1 expression at any time. To definitively locate the transcription initiation sites, we performed restriction-ligase-mediated 5' RACE, to selectively amplify only mRNA with a 5' cap. An initial characterization of the sequence upstream of the initiation sites determined six possible binding sites for the prostate specific regulatory protein NKX3.1 and four potential binding sites for the PPAR/RXR heterodimer that is involved in the control of cell differentiation and apoptosis.

Dyrithiopsis lakefuxianensis gen. et sp. nov. from Fuxian Lake, Yunnan, China, and notes on the taxonomic confusion surrounding Dyrithium.

A new taxon with Dyrithium-like characteristics was collected from Lake Fuxian in China. The taxon is typical of the Amphisphaeriaceae in that it has relatively large, ostiolate, immersed ascomata, unitunicate asci with a J+ subapical ring, and brown ascospores. It is similar to Dyrithium in that it has muriform ascospores, but considerable confusion surrounds this genus. In Dyrithium asci are bitunicate and lack a J+ subapical ring, while this was not true of our species. A new genus, Dyrithiopsis, therefore is established to accommodate this new taxon. Details of its anamorph also are provided, based on cultural studies. Parsimony analyses of part of the large-subunit rDNA provide further evidence to support the familial placement of this new genus in the Amphisphaeriaceae. The taxonomic position of Dyrithium also is discussed.