The factors influencing the occurrence of post-ischemic stroke depression and its relationship with the burden score of cerebral small vessel disease.

OBJECTIVE: This study explored the determinants of post-stroke depression (PSD) in ischemic stroke (AIS) patients and its association with the burden score of cerebral small vessel disease (CSVD). PATIENTS AND METHODS: We analyzed 374 AIS patients treated between January 2020 and January 2022. Patients were categorized into 90 with PSD and 284 without PSD, enabling an investigation into PSD risk factors and the CSVD-PSD relationship. RESULTS: There was no significant difference in health factors between PSD and non-PSD patients (p>0.05). However, significant disparities were noted in age, gender, initial Barthel Index (BI), Mini-Mental State Examination (MMSE) score, plasma fibrinogen, homocysteine, red cell distribution width, National Institutes of Health Stroke Scale (NIHSS) score, and CSVD burden score (p<0.05). Regression analysis indicated that these variables were pivotal PSD predictors (OR>1, p<0.05). Surprisingly, a positive correlation with PSD occurrence was found for age, NIHSS score, plasma fibrinogen, homocysteine levels, red cell distribution width, CSVD burden score (r=0.565, 0.615, 0.482, 0.514, 0.572, 0.608, respectively; p<0.05). Meanwhile, the MMSE score and BI index were inversely related to PSD onset (r=-0.604, -0.590; p<0.05). The ROC curve analysis of the combination model based on MMSE, NIHSS and CSVD score revealed an AUC of 0.926 and Youden's index of 0.744. CONCLUSIONS: Age, MMSE score, BI index, NIHSS score, plasma fibrinogen concentration, homocysteine level, red blood cell distribution width, and CSVD burden score are all major influencing factors in the occurrence of PSD. The combination model based on MMSE, NIHSS, and CSVD scores presented a valuable approach to predicting PSD.

Energy distribution of lost high-energy runaway electrons based on their bremsstrahlung emission in the EAST tokamak.

We report in this paper the study towards revealing the energy distribution of lost high-energy runaway electrons based on their bremsstrahlung emission. The high-energy hard x-rays are originated from the bremsstrahlung emission of lost runaway electrons in the experimental advanced superconducting tokamak (EAST) tokamak, and the energy spectra are measured using a gamma spectrometer. The energy distribution of the runaway electrons is reconstructed from this hard x-ray energy spectrum using a deconvolution algorithm. The results indicate that the energy distribution of the lost high-energy runaway electrons can be obtained with the deconvolution approach. In the specific case in this paper, the runaway electron energy was peaked around 8 MeV, covering from 6 MeV to 14 MeV.

Experimental examination of a method to estimate temporal effect by neutrons and γ-rays on scintillation light in scintillator-based soft x-ray diagnostic of experimental advanced superconducting tokamak and large helical device.

Scintillators, which are more tolerant of neutrons or γ-rays than semiconductors, are a promising candidate for soft X-ray (SX) diagnostics in high neutron flux environments such as JT-60SA or ITER. Although scintillators are tolerant of radiations, neutrons and γ-rays can cause scintillation light and become noise on SX signals. Therefore, a method to estimate the temporal effect by the radiations on SX signals and an appropriate design of the radiation shield based on the estimation are required. In previous studies, it has been proposed for estimating the effect by the radiations to calculate the absorption powers due to SXs, neutrons, and γ-rays in scintillators assuming that amplitudes of scintillation light are proportional to the absorption powers. In this study, an experimental examination of this proposal is conducted in the Experimental Advanced Superconducting Tokamak (EAST). It is shown that the proposal may be valid in the examination of EAST. In addition to results in EAST, initial results of a multi-channel scintillator-based SX diagnostic in the Large Helical Device (LHD) are introduced. Although a scintillator-based SX diagnostic in LHD observes oscillations of SXs by magnetohydrodynamic (MHD) phenomena successfully, the observed temporal effect on SX signals by neutrons or γ-rays is more significant than the expected effect, which is estimated by calculating the absorption powers. One of the possible reasons for the contradiction between the results in EAST and LHD is unexpected γ-rays around the scintillators in LHD. Although the temporal effect by the radiations is significant in the current system of LHD, the degradation of amplitudes of SX signals after the deuterium plasma experiments is not observed with the current level of the fluence. The scintillator-based SX diagnostic in LHD may work as a diagnostic to research MHD instabilities in deuterium plasma experiments without additional maintenance during an experimental campaign by making the pinhole larger or setting an additional radiation shield.

Intratumoral IL17-producing cells infiltration correlate with antitumor immune contexture and improved response to adjuvant chemotherapy in gastric cancer.

BACKGROUND: Tumor IL17-producing (IL17A+) cells infiltration has different prognostic values among various cancers. The objective of this study was to assess the effect of IL17A+ cells in gastric cancer. PATIENTS AND METHODS: The study included two patient cohorts, the Cancer Genome Atlas cohort (TCGA, n = 351) and the Zhongshan Hospital cohort (ZSHC, n = 458). The TCGA and ZSHC were used for mRNA-related and cells infiltration-related analyses, respectively. The roles of IL17A mRNA and IL17A+ cells in overall survival (OS), response to adjuvant chemotherapy (ACT), and immune contexture were evaluated. Another independent cohort was included to identify the correlation between mRNA of IL17A and IL17A+ cells infiltration (the preliminary Zhongshan Hospital cohort, PZSHC, n = 21). RESULTS: The infiltration of IL17A+ cells was positively correlated with the expression of IL17A mRNA (Spearman's ρ = 0.811; P < 0.001). High IL17A mRNA expression and intratumoral IL17A+ cells were correlated with improved OS and remained to be significant after adjusted for confounders. Patients with TNM II/III disease whose tumor present higher intratumoral IL17A+ cells or lower peritumoral IL17A+ cells can benefit more from ACT. Elevated IL17A mRNA expression and increased intratumoral IL17A+ cells infiltration was associated with more antitumor mast cells and nature killer cells infiltration and less pro-tumor M2 macrophages infiltration. High IL17A mRNA expression represented a Th17 cells signature and immune response process and was correlated with increased cytotoxic GZMA, GZMB, IFNG, PRF1, and TNFSF11 expression. CONCLUSIONS: IL17A mRNA expression and intratumoral IL17A+ cells infiltration were correlated with antitumor immune contexture. IL17A+ cells infiltration could be used as an independent prognostic biomarker for OS and predictive biomarker for superior response to ACT, and further prospective validation needs to be conducted.

Development of gamma ray spectrometer with high energy and time resolutions on EAST tokamak.

A new gamma ray spectrometer with high energy and time resolutions has been developed and installed on the EAST tokamak to study fast ion and runaway electron behaviors. The spectrometer is based on a LaBr3(Ce) scintillator detector and a fully digital data acquisition system that is based on a digitizer with digital pulse processing algorithms. The energy resolution of the spectrometer is about 3.9% at 662 keV, and the spectrometer can operate stably at a counting rate as high as 1 MHz, monitored by using a light emitting diode monitoring system. The measured gamma ray spectrum is simulated based on Geant4 and unfolded with the high-resolution boosted Gold deconvolution algorithm, aiming at reconstructing the energy distribution functions of fast ions and runaway electrons.

Performance of fast-ion loss diagnostic on EAST.

The scintillator-based detector for fast-ion loss measurements has been installed on EAST. To obtain high temporal resolution for fast-ion loss diagnostics, fast photomultiplier tube systems have been developed which can supply the complementary measurements to the previous image system with good energy and pitch resolution by using a CCD camera. By applying the rotatable platform, the prompt losses of beam-ions can be measured in normal and reverse magnetic field. The thick-target bremsstrahlung occurring in the stainless steel shield with energetic electrons can produce X-rays, which will strike on the scintillator based detector. To understand this interference on fast-ion loss signals, the effects of energetic electrons on the scintillator-based detector are studied, including runaway electrons in the plasma ramping-up phase and fast electrons accelerated by the lower hybrid wave.

First results from gamma ray diagnostics in EAST Tokamak.

Gamma ray diagnostics has been developed in the EAST tokamak recently. Six BGO scintillator detectors are arranged on the down-half cross-section and pointed at the up-half cross-section of plasma, with space resolution about 15 cm and energy range from 0.3 MeV to 6 MeV. Three main gamma ray peaks in the energy spectra have been observed and are identified as the results of nuclear reactions 207Pb(n, n')207mPb, H(n, γ) D, and D(p, γ)3He, respectively. Upgrading of the system is in progress by using LaBr3(Ce) scintillator, fast photo-multiplier tubes, and a fully digital data acquisition system based on high sample frequency digitizers with digital pulse processing algorithms.

Status of neutron diagnostics on the experimental advanced superconducting tokamak.

Neutron diagnostics have become a significant means to study energetic particles in high power auxiliary heating plasmas on the Experimental Advanced Superconducting Tokamak (EAST). Several kinds of neutron diagnostic systems have been implemented for time-resolved measurements of D-D neutron flux, fluctuation, emission profile, and spectrum. All detectors have been calibrated in laboratory, and in situ calibration using 252Cf neutron source in EAST is in preparation. A new technology of digitized pulse signal processing is adopted in a wide dynamic range neutron flux monitor, compact recoil proton spectrometer, and time of flight spectrometer. Improvements will be made continuously to the system to achieve better adaptation to the EAST's harsh γ-ray and electro-magnetic radiation environment.

First Report of Anthracnose of Malva sylvestris Caused by Colletotrichum trifolii in China.

Common mallow (Malva sylvestris L.) is a perennial medicinal plant in the Malvaceae family, which is native to Asia, Europe, and North Africa. In July 2012, typical symptoms of anthracnose disease, with a disease incidence of ~70%, were observed on common mallow in the Medicinal Herb Garden of Shenyang Pharmaceutical University, Liaoning, China. The fungus mainly infected the stalks and leaves of M. sylvestris. Pinpoint, brownish lesions initially appeared at the flowering stage and the disease spread within the field. The lesions on stems gradually enlarged and became dark brown, elliptical, and slightly concave. Concurrently, acervuli and mucilaginous conidial masses of the pathogen appeared on lesions under moist conditions. Conidia were hyaline, one-celled, cylindrical with both ends rounded, and measured 10.0 to 12.5 × 2.5 to 4.0 µm (mean 11.3 × 3.3 µm). The fungus was isolated from symptomatic tissues. Small pieces from leaves and stems were surface disinfested with 70% ethanol and 1.5% sodium hypochlorite for 1 min, then rinsed three times with sterile distilled water, and cultured on potato dextrose agar (PDA) at 25°C. The colonies on PDA had initially white aerial mycelia, and later became greenish black with regularly whorled rings. To confirm Koch's postulates, five 3-month-old plants of M. sylvestris were inoculated with a conidial suspension (105 conidia/ml) prepared from PDA cultures incubated for 14 days. Five non-inoculated plants served as controls. The plants were maintained in the greenhouse at 22 to 25°C and about 75% relative humidity under natural daylight. Typical symptoms on inoculated plants were reproduced after ~10 to 14 days, whereas control plants remained asymptomatic. The pathogen was successfully recovered from symptomatic tissues and re-identified, completing Koch's postulates. The internal transcribed spacer (ITS) and large subunit -28S (LSU) region of rDNA was amplified with primers ITS1/ITS4 and NL1/NL4, respectively, and sequenced. Phylogenetic trees (ITS and LSU) that were obtained using MEGE3.1 with the neighbor-joining method showed that both of the isolates fall in the Colletotrichum trifolii clade. The representative sequences (ITS and LSU) were deposited in GenBank (Accession Nos. KJ155692 and KJ920935). The fungus isolated from symptomatic tissues was identified as C. trifolii on the basis of morphological, cultural characteristics, and sequence analysis (2). According to previous references, C. orbiculare and C. malvarum on Malvaceae were respectively described in America and Europe (1,3,4). However, the isolate from M. sylvestris significantly differed from those of C. orbiculare and C. malvarum in cultural characteristics and sequence analysis. In this paper, the results showed that M. sylvestris is a new host of C. trifolii. To our knowledge, this is the first report of mallow anthracnose caused by C. trifolii in China. References: (1) J. A. Bailey et al. Phytopathology 86:1076, 1996. (2) U. Damm et al. Fungal Divers. 61:29, 2013. (3) K. Hyde et al. Fungal Divers. 39:147, 2009. (4) L. Tosi et al. Plant Dis. 88:425, 2004.

First Report of Sclerotinia Rot of Chinese Atractylodes Caused by Sclerotinia nivalis in China.

Chinese atractylodes (Atractylodes japonica Koidz.ez Kitam.) is a perennial herb in the Compositae family, and is widely distributed in China. The dried rhizomes of the plant are used in traditional Chinese medicine. During the summer of 2011, typical signs and symptoms of Sclerotinia rot were observed on Chinese atractylodes in a production field of Liaoning Province of China. Symptoms were observed in plants at the flowering stage, distributed in patches throughout the rows, and with a disease incidence of approximately 10 to 15%. The lower mature leaves of infected plants first became yellow and wilted, basal stem areas showed a black-brown rot at the same time under conditions of high humidity, and white cottony mycelium formed along the basal stem and soil surfaces. Ultimately, the basal stem and roots rotted and the plants wilted and died quickly. Black, irregular sclerotia (average 0.8 to 6.9 mm in diameter) were also observed within the pith cavity of split stems and rotted roots. The pathogen was isolated from symptomatic tissues and sclerotia, surface disinfested with 2% sodium hypochlorite, and cultured on potato dextrose agar (PDA) (1). The fungus was mesophilic, with an optimum temperature for mycelial growth in culture of about 20°C. Colonies on PDA produced masses of white aerial mycelium, with small white flocci distributed among sclerotia. After 2 weeks, sclerotia 0.5 to 4.5 mm in diameter were produced near the margin in a uniform distribution. Sclerotia were spherical, elongated, or fused to form irregular shapes and tightly attached to the agar surface by their under surface, which could be seen through the bottom of the petri dishes. DNA sequences of five replicates were obtained using the TianGen DNA secure plant kit. The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1/ITS4 and sequenced. BLAST analysis of the 513-bp segment showed high similarity (99%) with a sequence of Sclerotinia nivalis (GenBank Accession No. AB516670). A representative sequence was deposited in GenBank (Accession No. JX294862). The fungus isolated from symptomatic tissues was identified as S. nivalis Saito on the basis of morphological and cultural characteristics (2,3) and ITS sequence analysis. Symptoms were reproduced in the greenhouse by inoculating the basal stem and roots of 15 atractylodes plants at the 7- to 10-leaf stage. Inoculum was prepared by macerating 14-day-old PDA cultures of the fungus in a blender and placing the mixture (approximately 20 g) into the potting medium of each plant. Sterile PDA was used to inoculate the five control plants. Plants were maintained in a greenhouse at 22 to 25°C and about 75% relative humidity. After 7 to 10 days, symptoms were similar to those in the fields. Lower leaves of inoculated plants became yellow and wilted, and infected plants died 2 weeks after inoculation, whereas control plants remained healthy. The pathogen was successfully recovered from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report of Sclerotinia rot of Chinese atractylodes. Given its wide host range, S. nivalis has great potential to become an economically important plant pathogen. References: (1) W. G. Kim and W. D. Cho. Mycobiology 30:41, 2002. (2) G. Q. Li et al. Mycol. Res. 104:232, 2000. (3) I. Saito. Mycoscience 38:227, 1997.

Sclerotinia Rot of Windflower (Pulsatilla koreana Nakai) Caused by Sclerotinia nivalis in China.

Windflowers (Pulsatilla spp.) are perennial medicinal plants in the family Ranunculaceae with high economic as well as medicinal value in China. It is a commonly used traditional Chinese medicine (1). In 2012, a stem rot disease was observed on windflower (P. koreana Nakai) at flowering stages in fields of Liaoning Province, China. Disease incidence ranged from 10 to 65% (average of 45%) and resulted in approximately 25 to 60% yield loss. The infected area of plants initially takes on a dark green or brown water-soaked appearance and then becomes paler. Soon after, plants turn brown or black and die. If moisture conditions remain conducive, a cottony mycelium cover was observed on the affected area. Later, the mycelium usually produces numerous black sclerotia that were up to 1 cm long in affected plant parts. Diseased tissue were surface sterilized for 1 min in 1% NaOCl (v/v), rinsed with sterilized distilled water, and plated on potato dextrose agar (PDA). Isolates were mesophilic, with an optimum growth temperature of around 20°C. Colonies on PDA were white, with abundant aerial mycelium. Some mycelium in the colony center, especially that submerged in the medium, was light brown. Sclerotia were spherical to subspherical, elongated or fused to form irregular shapes, 2.5 to 9.0 × 2.0 to 6.8 mm. They were tightly attached to the agar surface by their under surface, which could be seen through the bottom of the petri dishes. These characteristics were consistent with Sclerotinia nivalis (2,3). Two isolates were selected for molecular identification. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (4) and sequenced. The obtained sequences (GenBank Accession Nos. JX206828 and JX424615) showed 99 and 100% homology with the sequences of S. nivalis in GenBank (Accession No. AB516670). To demonstrate pathogenicity, mycelial blocks of the isolates grown on PDA were placed on the base of the stems of ten 1-month-old Pulsatilla koreana plants. The same numbers of control plants were treated with PDA plugs as a control. The inoculated plants were incubated at 25°C with a 12-h photoperiod. After 4 days, the initiation of stem necrosis was observed, and 9 days after inoculation, the plants collapsed and died. Control plants had no symptoms. The same fungus was reisolated from all inoculated plants, satisfying Koch's postulates. To our knowledge, this is the first confirmed report of windflower as a natural host for S. nivalis in China. References: (1) S. C. Bang et al. J. Nat. Prod. 68:268, 2005. (2) G. Q. Li et al. Mycol. Res. 104:232, 2000. (3) I. Saito. Mycoscience. 38:227, 1997. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Severe Outbreak of Rust Caused by Coleosporium pulsatillae Detected on Pulsatilla spp. in China.

Windflower (Pulsatilla spp.) is a perennial medicinal plant in the Ranunculaceae with high economic value as well as medicinal value in China. It is a commonly used Chinese herbal medicine. In 2007, two species, Pulsatilla koreana Nakai and P. chinensis Regel, were observed with severe rust symptoms at three locations (Shenyang City, Benxi City, and Fushun County) in Liaoning, China. The diseased area was estimated to be more than 500 ha and the yield was reduced by 30% on average with up to 65% yield losses in some fields. Since its first record in 2007, the disease has been recorded every year in parts of China. A survey of all cultivated fields in August 2011 revealed that 90% of the Pulsatilla plants were heavily infected with rust. Early symptoms on the adaxial leaf surfaces were small, circular, yellow spots that later enlarged, coalesced, and developed necrotic centers. On the abaxial side, numerous yellow rust uredinia were visible. Urediniospores are globose or ellipsoidal, sometimes angular in shape, with a yellowish content, and measured 22.6 to 39.4 × 15.2 to 23.9 µm. Spore walls were coarsely verrucose when examined by light and scanning electron microscopy. Telia and teliospores were observed in mid-August. Hypophyllous telia often formed around uredinial clusters. Telia were 0.3 to 1.1 mm wide and erumpent with numerous teliospores in a compact layer. Teliospores were clavate, oblong to ellipsoidal, 60.2 to 120.8 × 12.1 to 24.4 µm, gelatinous, and one celled. Teliospores were four celled with the division of the protoplast into an internal four-celled basidium. Pathogenicity tests included inoculation of young P. koreana plants by spraying a urediniospore suspension (30,000 spores/ml of deionized water). Inoculated plants were incubated at 25°C for 48 h, and typical rust symptoms and uredinia developed in 10 to 15 days. On the basis of symptomatology, the host, and morphology of uredinial and telia, the fungus was identified as Coleosporium pulsatillae (Str.) Lév (2). The internal transcribed spacer (ITS) region was amplified using ITS1-F and ITS4-B primers (3), and an amplified product of 817 bp, specific for the species C. pulsatillae, was obtained. The sequence was deposited in GenBank (Accession No. JQ029765). Although this pathogen has been mentioned as part of a fungal species survey from China, it was not fully described (4). This pathogen has been reported on Anemone chinensis in Austria, Sibiria (2), and Korea (1). It has not been reported from anywhere else in China. To our knowledge, this is the first fully described record and most severe outbreak of C. pulsatillae on windflower. References: (1) W. D. Cho and H. D. Shin. List of Plant Diseases in Korea. Korean Society of Plant Pathology, Seoul, Korea, 2004. (2) J. B. De-Toni. Sylloge Fungorum 7:754, 1888. (3) M. Gardes et al. Mol. Ecol. 2:113, 1993. (4) F. L. Tai. Sylloge Fungorum Sinicorum. Science Press, Beijing, 1979.

Differential expression profiling of ovarian genes in prelaying and laying geese.

The identification of the differential expression of genes in the ovaries of egg-laying and prelaying Zi geese is required to improve the laying performance of the geese. In the present study, suppression subtractive hybridization and reverse dot-blot were employed to identify such genes, using the ovary as a model. Furthermore, expression profiling of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 in ovaries from geese was performed by quantitative real-time PCR. Total RNA from the ovaries of laying and prelaying Zi geese was pooled and the mRNA was isolated. The cDNA that was reverse-transcribed from the ovarian mRNA of the prelaying geese was subtracted from the cDNA isolated from the laying geese. Four hundred sixty-five clones containing putative differentially expressed gene fragments were further identified by reverse dot-blot. Ninety-seven clones were subjected to sequencing and further analysis. Sequence analysis showed that the expression of 18 known (including a mitochondrial gene) and 8 unknown gene fragments was higher in the ovaries of laying geese compared with prelaying geese. Seventeen of the known genes encode proteins that belong to groups involved with binding, catalytic activity, enzyme regulatory activity, signal transducer activity, structural molecule, and transporter activity. The results of the quantitative real-time PCR showed that the expression of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 was higher in the ovaries of the laying geese than in those of the prelaying geese (P<0.05). These differentially expressed genes may be relevant to the progression of prelaying geese to the egg-laying stage. Further study is required to elucidate the molecular mechanism that controls egg-laying in geese, to improve the productivity of laying geese.

Role of Fas/FasL genes in azoospermia or oligozoospermia induced by testosterone undecanoate in rhesus monkey.

AIM: To investigate role of apoptosis related genes Fas/FasL in azoospermia or oligozoospermia induced by testosterone undecanoate. METHODS: TUNEL was used to detect the apoptotic signal of testicular cells. Immunohistochemistry and Western blot were used to quantitatively or qualitatively analyze the expression of these apoptosis-related proteins. RESULTS: After injection of testosterone undecanoate, both the apoptotic signal in germ cells and the expression of Fas/FasL in testis increased correlatively in a time-dependent manner, reaching a maximum level on day 30. CONCLUSION: Fas system may initiate and regulate the germ cell apoptosis induced by testosterone undecanoate.

[The cloning and verification of the Sox9 gene of the rice field eel].

The Sox 9 gene plays important roles in sex determination, which is conserved in a variety of species including mammals and chicken. The positive lambda clone of the Sox 9 gene of the rice field eel was analyzed by restriction enzymes and the restriction map was constructed. This clone was further verified to be the Sox9 gene by PCR amplification and sequencing of the HMG box region. The cloning of the Sox9 gene of the rice field eel further suggests the evolutionary conservation of this gene.

[Primed in situ labelling of human metaphase chromosomes].

Primed in situ labelling (PRINS) contributes to the elucidation of the organization of eukaryotic chromosomes and provides an alternative way to localize gene on chromosomes. Here we present the results of random primer and SOX degenerate primer PRINS on human metaphase chromosomes. The particular band pattern of dark and light was observed by using random primer PRINS. SOX degenerate primer PRINS revealed more SOX gene loci and proved further that SOX genes were not clustered on human chromosomes.

[Painting the chromosomes of fishes with human sex chromosome-specific DNA probes].

Chromosome painting is one of the effective methods in studying the evolution of genomes. The chromosomes of the species of fishes--M. aculeatus, M. albus and D. rerio were painted respectively with human X and Y chromosome-specific DNA probes. The results showed that some homologous segments of human X chromosome were found in the genomes of all the three kind of fishes, all the segments were scattered on several pairs of homologous chromosomes in each kind of fish. But no homologous segment of the human Y chromosome was found.

Expression of Hsp70-2 in unilateral cryptorchid testis of rhesus monkey during germ cell apoptosis.

We investigated the possible role of Hsp70-2 in germ cell apoptosis induced by heat stress in monkey unilateral cryptorchid testis. The study focused on in situ analysis of the testicular cell DNA fragmentation and on the possible relationship between Hsp70-2 expression and germ cell apoptosis. The TUNEL result showed that most of the germ cells were labeled in the cryptorchid testis on d 5 after induction of cryptorchidism; that with most of the apoptotic germ cells depleted, only a few germ cells were labeled on d 10; and that almost no apoptotic signal was observed in the cryptorchid testis on d 15 and thereafter. This indicates that the increasing germ cell degeneration in cryptorchid testis may take the form of apoptosis. Using in situ hybridization, immunohistochemistry, and Northern blot, we examined the changes of Hsp70-2 expression in the monkey cryptorchid testis. The level of Hsp70-2 mRNA decreased slightly, while the expression of HSP70-2 protein was almost unchanged at the early stage of germ cell apoptosis in the cryptorchid testis on d 5 and dropped dramatically along with the loss of apoptotic germ cells in the cryptorchid testis on d 10 after operation. It is therefore suggested that Hsp70-2 might not take part in inhibiting the apoptosis of germ cells at the early stage during operation-induced cryptorchid testis, and that Hsp70-2 gene does not belong to the immediate early related gene responsible for germ cell apoptosis induced by heat stress.

[Transgenic loach produced by using sperm cells mediated by high molecules].

The pCH110 plasmid DNA which has a marker gene LacZ controlled by a SV40 promoter was mixed with a kind of high molecule of dendrimers(cyclic core dendrimer) and then formed a DNA/dendrimer complex. Loach sperms were incubated with the DNA/dendrimer complex. Then in situ hybridization was used to detect whether the sperm cells captured the foreign DNA or not. Results show that the number of sperm capturing foreign DNA via dendrimers as vectors was obviously increased. The sperms were mixed with eggs for in vitro fertilization. PCR was used to detect the transgenic loach fries. Obvious expression of LacZ gene in the head of transgenic loach were observed by analysis through LacZ histochemical staining of embryos or fries.

[The chromosome mapping of PdSox8 and PdSox9 in two kinds of loaches].

With the Dig-labelled PdSox8 and PdSox9 as probes, the chromosome mapping of Sox8 and Sox9 in Paramisgurnus dabryanus and Misgurnus anguillicaudatus were analyzed by chromosome in situ hybridization technique. In P. dabryanus, the PdSox8 and PdSox9 were successfully localized in the No. 4 and No. 2 telocentric chromosome respectively, the relative distance to centromere is 40.2% and 67.5%; In M. anguillicaudatus, they were localized in the No. 9 and No. 6 telocentric chromosome respectively, the relative distance to centromere is 58.3% and 30.8%.