An aquaporin and an aquaglyceroporin have roles in low temperature adaptation of mosquitoes (Anopheles sinensis).

Mosquitoes (Anopheles sinensis), widely geographically distributed in Asia including China, are the primary vector of the malaria parasite Plasmodium vivax and other parasitic diseases such as Malayan filariasis. An. sinensis can survive through low winter temperatures. Aquaporin channels are found in all life forms, where they facilitate environmental adaptation by allowing rapid trans-cellular movement of water (classical aquaporins) or water and solutes such as glycerol (aquaglyceroporins). Here, we identified and characterized 2 aquaporin (AQP) homologs in An. sinensis: AsAQP2 (An. sinensis aquaglyceroporin) and AsAQP4 (An. sinensis aquaporin). When expressed in frog (Xenopus laevis) oocytes, AsAQP2 transported water, glycerol, and urea; AsAQP4 transported only water. Water permeation through AsAQP2 and AsAQP4 was inhibited by mercuric chloride. AsAQP2 expression was slightly higher in adult female mosquitoes than in males, and AsAQP4 expression was significantly higher in adult males. The 2 AsAQPs were highly expressed in Malpighian tubules and midgut. AsAQP2 and AsAQP4 expression was up-regulated by blood feeding compared with sugar feeding. At freezing point (0 °C), the AsAQP4 expression level increased and An. sinensis survival time reduced compared with those at normal temperature (26 °C). At low temperature (8 °C), the AsAQP2 and AsAQP4 expression levels decreased and survival time was significantly longer compared with those at 26 °C. These results suggest that AsAQP2 and AsAQP4 have roles in water homeostasis during blood digestion and in low temperature adaptation of A. sinensis. Together, our results show that the 2 AQPs are important for mosquito diuresis after blood feeding and when exposed to low temperatures.

Male reproductive system and simulated high-altitude environment: preliminary results in rats.

This study assessed the effects of a simulated high-altitude environment on the reproductive system of prepubertal male rats and the reversibility of these effects upon return to a normal environment. Three-week-old male Wistar rats were randomly allocated to 4 groups that were exposed to different conditions: a normal environment for 6 weeks and 12 weeks, respectively, hypobaric hypoxia for 6 weeks, and hypobaric hypoxia for 6 weeks followed by a normal environment for 6 weeks. Multiple pathophysiological parameters were evaluated at the histological, endocrine, and molecular levels. Hypobaric hypoxia exposure for 6 weeks during the prepubertal phase significantly altered physiological parameters, body functions, blood indices, and reproductive potential. Six weeks after returning to a normal environment, the damaged reproductive functions partially recovered due to compensatory mechanisms. However, several changes were not reversed after returning to a normal environment for 6 weeks, including disorders of body development and metabolism, increased red blood cells, increased fasting blood glucose, abnormal blood lipid metabolism, decreased testicular and epididymis weights, abnormal reproductive hormone levels, excessive apoptosis of reproductive cells, and decreased sperm concentration. In summary, a hypobaric hypoxic environment significantly impaired the reproductive function of prepubertal male rats, and a return to normal conditions during the postpubertal phase did not fully recover these impairments.

[Ginsenoside Rg3 Regulates Cisplatin Resistance in Gastric Cancer by Wnt/ß-catenin Signaling Pathway].

Objective To investigate the inhibitory effect of ginsenoside Rg3 combined with cisplatin (DDP) on DDP-resistant cell line SGC-7901/DDP and their molecular mechanism.Methods SGC-7901/DDP cells were divided into four groups including a control group,a ginsenoside Rg3 (40 µg/ml) treatment group,a DDP (1.40 µg/ml) treatment group,and a drug combination treatment group.The proliferation ability of SGC-7901/DDP cells was detected by MTT,EdU,and colony formation assays.The apoptosis ability of SGC-7901/DDP cell was detected by flow cytometry and Hoechst 33342 staining.The protein levels of apoptosis-related markers were detected by Western blotting.The migration ability of SGC-7901/DDP cells was detected by wound healing and Transwell assays.The expression levels of proteins in epithelial-mesenchymal transformation (EMT) and Wnt/ß-catenin signaling pathway were determined by Western blotting and immunofluorescence staining.Results Compared with the ginsenoside Rg3 or the DDP treatment groups,the drug combination treatment group inhibited the proliferation (t=8.062,P=0.001;t=7.090,P=0.002),colony formation (t=8.062,P=0.001;t=6.144,P=0.004),and migration (t=7.424,P=0.002;t=4.317,P=0.013),and promoted the apoptosis (t=5.530,P=0.031;t=6.036,P=0.026) of SGC-7901/DDP cells.Compared with the ginsenoside Rg3 and the DDP treatment groups,the drug combination treatment group down-regulated the expression levels of EMT-associated proteins including vimentin (t=24.450,P<0.001;t=14.750,P<0.001),Snail (t=29.640,P<0.001;t=70.700,P<0.001),Slug (t=89.230,P<0.001;t=87.360,P<0.001),matrix metalloproteinase (MMP) 2 (t=84.540,P<0.001;t=67.120,P<0.001),and MMP9 (t=19.010,P<0.001;t=10.890,P<0.001),as well as those of Wnt/ß-catenin signaling pathway related proteins including Wnt (t=35.480,P<0.001;t=14.670,P<0.001),ß-catenin (t=155.800,P<0.001;t=118.100,P<0.001),C-myc (t=20.870,P<0.001;t=3.334,P=0.029),and cyclin D1 (t=5.007,P=0.008;t=8.347,P=0.001).Meanwhile,it up-regulated the expression of epithelial cells including E-cadherin (t=36.450,P<0.001;t=33.810,P<0.001) and ZO-1 (t=37.060,P<0.001;t=37.030,P<0.001).Conclusion Ginsenoside Rg3 enhanced the sensitivity of SGC-7901/DDP cells to DDP by inhibiting the activity of Wnt/ß-catenin signaling pathway.

Exploring cryptic biodiversity in a world heritage site: a new pitviper (Squamata, Viperidae, Crotalinae) from Jiuzhaigou, Aba, Sichuan, China.

This study presents a comprehensive morphological comparison along with molecular phylogeny of the genus Gloydius based on five mitochondrial genes (12S, 16S, COI, cytb, and ND4). The specimens collected from Jiuzhaigou National Nature Reserve are shown to be a new species, Gloydiuslateralis sp. nov. Zhang, Shi, Jiang & Shi based on a combination of morphological and molecular accounts. G.lateralis sp. nov. differs from other congeneric species by a series of diagnostic morphological characteristics and forms a strongly supported monophyletic group. The new species is phylogenetically closely related to G.swild, another recently described species from Heishui, Aba, Sichuan.

Asymptomatic traumatic rupture of an intracranial dermoid cyst: A case report.

BACKGROUND: Previous studies reported that most of the intracranial dermoid cyst ruptures were spontaneous, and only a few were traumatic, with asymptomatic much rarer than the symptomatic ruptures. Hence, how to deal with the asymptomatic traumatic rupture of intracranial dermoid cyst remains a challenge in the clinic. CASE SUMMARY: A 59-year-old man was accidentally diagnosed with intracranial dermoid cyst through a cranial computed tomography (CT) scan due to a car accident. A mixed-density lesion with fat and a calcified margin was observed in the midline of the posterior fossa, accompanied with lipid droplet drifts in brain sulci, fissures, cisterns, and ventricles. After 1 wk of conservative observation, no change was observed on the updated cranial CT scan. After 2 wk of conservative observation, magnetic resonance imaging examination confirmed that the lesion was a traumatic rupture of a posterior fossa dermoid cyst with lipid droplet drifts. As the patient exhibited no adverse symptoms throughout the 2 wk, a 6-mo follow-up visit was arranged for him instead of aggressive treatment. Nonetheless, the patient did not show any abnormal neurological symptoms in the 6 mo of follow-up visits. CONCLUSION: Asymptomatic traumatic rupture of intracranial dermoid cyst could be just followed or treated conservatively rather than treated aggressively.

Sperm Parameters, ASAs and Apoptosis After Processing by the Double Tube and Swim up Methods.

The aim of this study was to improve the quality of semen samples by using a novel double-tube (DT) method. The DT method was developed to select sperm and compared with traditional swim-up (SU) technique for 31 semen samples. Sperm DNA integrity were tested with TUNEL and SCSA. Content of antisperm antibodies (ASA) in the semen was measured by ELISA and MAR. Levels of the caspase-3 in the sperm were assessed by western blotting. After SU and DT, 15 couples and 16 couples were underwent IVF-ET. The number of RCDs, the percentage of SDF and DFI, ASA and the level of caspase-3 were significantly decreased after DT and SU (p = .001 and p< .001). When the DT and SU compared, there were significant changes in the number of RCD, the percentage of SDF and DFI, ASA and the level of caspase-3 (p< 0.05-0.001). There was a higher cleavage rate (p = .017) and a lower abortion rate (p< .05) in DT-IVF group than in SU-IVF group. DT selection yielded spermatozoa with low RCDs, DFI, ASA, and caspase-3 which would be benefit for ART.

Classification of induced malaria case in an elimination setting: investigation of transfusion-transmitted malaria cases.

BACKGROUND: Since the National Malaria Elimination Action Plan was launched in China in 2010, local malaria transmission has decreased rapidly. Zero indigenous cases were reported since 2017. However, after 2010, the proportion of imported cases in China increased from 45.7% in 2010 to 99.9% in 2016, and almost all provinces of China have reported imported cases in recent years. Prevention of the reintroduction of malaria into China is crucial for the maintenance of its malaria-free status. Hence, it is of utmost importance to correctly identify the source of malaria infections within the country. CASE INTRODUCTION AND RESPONSE: In 2016 and 2017, three laboratory-confirmed cases of malaria caused by Plasmodium falciparum were identified in patients with no previous travel history to endemic areas were reported in Jiangsu Province, China, where malaria due to P. falciparum was eliminated about 30 years ago. These were diagnosed after 41, 31 and 39 days of seeking treatment, respectively, and all of them had received blood transfusions. Further investigations indicated that two of the cases had received blood from foreign students (from Indonesia and Ghana), and the other had received blood from an individual who had worked in Equatorial Guinea. All three blood donors were traced, and found to be carrying asymptomatic P. falciparum infections by microscopic examination and PCR. Furthermore, five polymorphic microsatellite markers (C1M4, C4M62, C13M13, C14M17, and C13M63) were typed and used to link parasites from the donors with those of the transfusion-receiving patients. CONCLUSIONS: Three transfusion-transmitted malaria cases were identified in China, all of which were due to the transfusion of blood donated by individuals who had contracted malaria outside the country. These cases can provide a reference for those faced with similar challenges in malaria case identification and classification in other regions. In addition, a stricter screening policy including the use of appropriate detection methods for malaria parasites should be developed and adopted for blood donation in regions undergoing malaria elimination.

Biologic response of sperm and seminal plasma to transient testicular heating.

Currently, there are few male contraceptive methods that are purely based on prevention of the entry of the sperm into the female reproductive tract. An alternative approach for designing reversible male contraceptive is achieved by transient testicular heating (TTH). This treatment, through massive germ cell apoptosis, causes reversible oligospermia or azoospermia. Here, we describe as how TTH causes DNA damage, oxidative stress, apoptosis, autophagy, sperm protein expression, and alters the biochemical components of seminal plasma. Further understanding of TTH will help design safe and reversible male contraception.

Osteology of Batrachuperus yenyuanensis (Urodela, Hynobiidae), a high-altitude mountain stream salamander from western China.

Batrachuperus yenyuanensis, commonly known as Yenyuan Stream Salamander, is a hynobiid species inhabiting high-altitude (2440-4025 m above sea level) mountain stream and pond environments along the eastern fringe of the Qinghai-Tibetan Plateau in western Sichuan Province, China. Although the species has been known for almost 70 years since its initial discovery in 1950, a thorough osteological description has never been provided. Our study provides a detailed account of the bony anatomy of this species, based on micro computed tomography scanning of multiple specimens collected from the type locality Shuangertang at Bailinshan, Yanyuan County, and several other localities in Sichuan Province. Our revised species diagnosis utilizes both bony and soft anatomical features. Comparative study of the specimens from the type locality in Yanyuan area with those from the nearby Xichang and Mianning areas confirms that they all pertain to Batrachuperus yenyuanensis, thereby removing doubt on the occurrence of the species in the latter areas. With this confirmation, the distribution of the species is extended from the type locality northwards some 180 km to the Mianning area, on both the west and east sides of the Yalong River, which is a major tributary of the upper Yangtze River. This distribution pattern indicates that the biogeographic origin and historical evolution of the species are closely associated with the orogeny of the Hengduan Mountains and formation of the Yalong River. Given the basalmost position of Batrachuperus yenyuanensis in relation to other congeneric species based on molecular studies, however, early expansion of the species distribution by dispersal is expected following the origin of the genus in early-middle Miocene in western Sichuan Province. Thus, the species may well have achieved its current distribution in western Sichuan before the drastic uplift of the Qinghai-Tibetan Plateau in Pliocene.

[Analysis of Correlation of Immune T Cell Subsets, TNF-α, IFN-γ and sFas levels with Severity and Prognosis in Aplastic Anemia Patients].

OBJECTIVE: To explore the correlation of immune T cell subsets and expression levels of TNF-α, IFN-γ and sFas with the severity and prognosis of aplastic anemia(AA) patients. METHODS: Ninety-five patients with aplastic anemia treated in our hospital were selected and enrolled in AA group, 43 cases of which were included in the acute group and 52 cases in the chronic group according to the disease status of patient's. Then, 50 cases of healthy volunteers in the same period were enrolled in the control group, and the changes of immune T cell subsets, and sFas serum TNF-α, IFN-γ and levels of the 3 groups were compared. The ROC curve was used to analyze the value of each index for evaluating the severity of the disease, 95 patients with aplastic anemia were followed up and the Kaplan-Miere survival curve was used to analyze the effect of different index levels on the progression-free survival of the patients. RESULTS: the serum level of CD4+ level and CD4+ /CD8+ ratio in AA group were significantly lower than those in the control group (P<0.05), and the CD8+ level in AA group was significantly higher than that in the control group (P<0.05). The CD4+ level and CD4+ /CD8+ ratio in the acute group was significantly lower than those in the chronic group (P<0.05), and the CD8+ level in the acute group was significantly higher than that in the chronic group (P<0.05). The serum level of TNF-α and IFN-γ of the patients in AA group was significantly higher than those in the control group (P<0.05), and the level of sFas in AA group was significantly lower than those in the control group (P<0.05). The levels of serum TNF-α and IFN-γ in the acute group were significantly higher than that in the chronic group (P<0.05), and the level of sFas was significantly lower than that in the chronic group (P<0.05). The analysis of each index value for predicting the severity of AA by ROC showed that the area under curve of CD4+ /CD8+ , TNF-α, IFN-γ and sFas for predicting the patients' condition was 0.954, 0.763, 0.853 and 0.857, respectively. Kaplan-Miere survival curve analysis showed that CD4+ /CD8+ ratio had a significant effect on progression-free survival (P<0.05), the TNF-α, IFN-γ and sFas levels had no significant effect on progression-free survival (P>0.05). CONCLUSION: The patients with aplastic anemia have significantly abnormal T lymphocyte subsets, there is a certain correlation between the serum TNF-α, IFN-γ and sFas levels with aplastic anemia, each index can reflect the disease severity, and the changes of T lymphocyte subsets has important influence on the clinical prognosis of the patients.

Effect of scrotal heating on sperm quality, seminal biochemical substances, and reproductive hormones in human fertile men.

At present, male contraceptive methods are only vasectomy and condoms, so it is necessary to research on male contraceptive techniques. The aim of this study is to observe the effects of scrotal heating (SH) on semen parameters, seminal l-carnitine (LC), epidermal growth factor (EGF), macrophage migration inhibitory factor (MIF), reproductive hormones and sperm chromosome numbers of adult healthy men, and to provide the experimental data for male contraception. The scrotums of 30 healthy male volunteers were exposed to the condition of 40 to 43°C SH belt warming 40 minutes each day for successive 2 days per week. The course of SH was continuous for 3 months. Computer-assisted semen analysis and hypo-osmotic swelling test, sperm DNA integrity, l-carnitine, MIF and EGF, and sperm fluorescence in situ hybridization were performed before, during, and after SH. The serum level of follicle stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) were measured by chemiluminescent immunoassay. The mean parameters of sperm concentration, vitality, and normal morphological sperm were significantly decreased in groups with sperms being collected during 1, 2, and 3 months of SH when compared with those in groups of pre-SH (P < 0.01). Statistically significant differences of sperm DNA fragmentation, normal sperm membrane functionality, levels of LC and MIF in semen, and LH, FSH, and T in serum were observed between the groups of before SH and after SH 3 months and the groups of during SH 1, 2, and 3 months (P < 0.001). The total rate of chromosome number for 13, 18, 21, X, and Y in the 3 months of SH was 13.7-fold greater (13.72%/1.69%) than before SH (P < 0.001). The constant SH can impact the semen quality, sperm DNA integrity, sperm chromosome, LC and MIF, and LH, FSH, and T in serum. Transient SH may be a new method for male contraception.

[Yam polysaccharide improves sperm viability and protects sperm DNA integrity in vitro].

OBJECTIVE: To clarify the roles of yam polysaccharide (YPS) in improving sperm viability and protecting sperm DNA integrity in vitro and provide a new approach to the treatment of oligoasthenozoospermia. METHODS: We collected samples by masturbation from 36 normal fertile males aged 27－39 years. Each sample was divided into six groups: blank control or treated with normal saline, vitamin C solution, and YPS solution at low (0.25 mg/ml), medium (1.0 mg/ml) or high concentration (5.0 mg/ml). Using eosin-Y staining, sperm hypotonic swelling (HOS) and sperm chromatin diffusion (SCD) test, we observed the effects of different concentrations of YPS on sperm viability, membrane integrity and nuclear DNA. RESULTS: After 24 and 48 hours of treatment, sperm viability was markedly reduced in the vitamin C (ï¼»28.5 ± 3.1ï¼½ and ï¼»6.5 ± 1.2ï¼½%), low-YPS (ï¼»31.3 ± 3.5ï¼½ and ï¼»6.5 ± 2.2ï¼½%), medium-YPS (ï¼»37.1 ± 3.5ï¼½ and ï¼»9.5 ± 2.8ï¼½%) and high-YPS groups (ï¼»38.3 ± 3.3ï¼½ and ï¼»9.0 ± 3.2ï¼½%) as compared with the blank control (ï¼»17.3 ± 2.1ï¼½ and ï¼»3.2 ± 1.3ï¼½%) (P <0.01) and normal saline groups (ï¼»13.4 ± 4.1ï¼½ and ï¼»3.1 ± 2.0ï¼½%) (P <0.01), and it was significantly higher in the medium- and high-YPS than in the vitamin C group (P <0.05 and P <0.01). The rate of sperm DNA fragmentation was remarkably decreased at 48 hours in the vitamin C (ï¼»30.5 ± 3.1ï¼½%), low-YPS (ï¼»29.4 ± 2.6ï¼½%), medium-YPS (ï¼»28.5 ± 2.3ï¼½%) and high-YPS groups (ï¼»27.9 ± 1.9ï¼½%) in comparison with the blank control (ï¼»41.7 ± 2.2ï¼½%) (P <0.01) and normal saline groups (ï¼»42.1 ± 3.3ï¼½%), markedly lower in the medium- and high-YPS than in the blank control, normal saline and vitamin C groups (P <0.05 or P <0.01), but with no statistically significant difference between the low-YPS and vitamin C groups (P >0.05). CONCLUSIONS: Yam polysaccharide can improve sperm viability and protect sperm DNA integrity in vitro.

Salvianolic Acid B Inhibits Aß Generation by Modulating BACE1 Activity in SH-SY5Y-APPsw Cells.

Alzheimer's disease (AD) is a neurodegenerative disease in humans. The accumulation of amyloid-ß (Aß) plays a critical role in the pathogenesis of AD. Previous studies indicated that Salvianolic acid B (SalB) could ameliorate Aß-induced memory impairment. However, whether SalB could influence the generation of Aß is unclear. Here, we show that SalB (25, 50, or 100 µM) reduces the generation of Aß40 and Aß42 in culture media by decreasing the protein expressions of BACE1 and sAPPß in SH-SY5Y-APPsw cells. Meanwhile, SalB increases the levels of ADAM10 and sAPPα in the cells. However, SalB has no impact on the protein expressions of APP and PS1. Moreover, SalB attenuates oxidative stress and inhibits the activity of GSK3ß, which might be related to the suppression of BACE1 expression and amyloidogenesis. Our study suggests that SalB is a promising therapeutic agent for AD by targeting Aß generation.

Changes in Levels of Seminal Nitric Oxide Synthase, Macrophage Migration Inhibitory Factor, Sperm DNA Integrity and Caspase-3 in Fertile Men after Scrotal Heat Stress.

BACKGROUND: This study observes changes in levels of seminal nitric oxide (NO), nitric oxide synthase (NOS), macrophage migration inhibitory factor (MIF), sperm DNA integrity, chromatin condensation and Caspase-3in adult healthy men after scrotal heat stress (SHS). METHODS: Exposure of the scrotum of 25 healthy male volunteers locally at 40-43°C SHS belt warming 40 min each day for successive 2 d per week. The course of SHS was continuously 3 months. Routine semen analysis, hypo-osmotic swelling (HOS) test, Aniline blue (AB) staining, HOS/AB and terminal deoxynucleotidyl transferase-mediated d UDP nick-end labeling (TUNEL) were carried out before, during and after SHS. Seminal NO and NOS contents were determined by nitrate reduction method. The activated Caspase-3 levels of spermatozoa and MIF in seminal plasma were measured by the enzyme-linked immunosorbent assay (ELISA) method. Statistical significance between mean values was determined using statistical ANOVA tests. RESULTS: The mean parameters of sperm concentration, motile and progressive motile sperm and normal morphological sperm were significantly decreased in groups during SHS 1, 2 and 3 months compared with those in groups of pre-SHS (P<0.001). Statistically significant differences of sperm DNA fragmentation, normal sperm membrane, and Caspase-3 activity as well as the level of NO, NOS and MIF in semen were observed between the groups before SHS and after SHS 3 months and the groups during SHS 1, 2 and 3 months (P<0.001). After three months of the SHS, various parameters recovered to the level before SHS. WBC in semen showed a positively significant correlation with the levels of NO, NOS, MIF and Caspase-3 activity. The percentage of abnormal sperm by using the test of HOS showed a positively significant correlation with that of HOS/AB. CONCLUSIONS: The continuously constant SHS can impact the semen quality and sperm DNA and chromatin, which may be contributed to the high level of NO, NOS, MIF and Caspase-3 by SHS.

[Progress in researches on molecular markers of Plasmodium falciparum drug resistance].

Effective chemotherapy is the mainstay of malaria control. However, it is undergoing the serious threat by resis- tance of falciparum malaria to antimalarial drugs. In recent years, with the development of molecular biology technology, molec- ular markers have been widely used to monitor antimalarial drug resistance. This paper reviews the researches on the common molecular markers related to Plasmodiumfalciparum drug resistance.

[Comparative study of assay methods for in vitro antimalarial drug efficacy testing in Plasmodium falciparum].

OBJECTIVE: To compare four assay methods of in vitro antimalarial drug efficacy testing, including WHO microtest, Plasmodial lactate dehydrogenase (pLDH), Histidine-rich protein II (HRP II) and SYBR Green I, so as to determine a stable, simple, rapid, and economic method for monitoring the drug sensitivity of malaria parasites and screening new antimalarial drugs. METHODS: WHO microtest, pLDH, HRP II and SYBR Green I were applied to test the drug efficacy of chloroquine, piperaquine and amodiaquine against four Plasmodiumfalciparum reference strains (3D7, FCC, K1 and Dd2), respectively. The consistency of the 50% inhibitory concentration (IC50) values from the four assay methods were analyzed by Friedman tests, Partial correlation analysis, Pearson' s correlation analysis and Bland-Altman plots. RESULTS: With the initial parasitemia ranged from 0.5% to 1%, there were no statistically significant differences (P> 0.05) among the IC50 values obtained by the four assay methods, which were correlated well (both P< 0.001). WHO microtest was highly labor-intensive, time-consuming and subjective; although HRP II was more sensitive than pLDH and SYBR Green I, which was more expensive; SYBR Green I was a simple, rapid and economic assay method. CONCLUSION: SYBR Green I, as a simple and cost-effective assay method, is suitable for high-throughput malaria drug sensitivity monitoring and research of new antimalarial drug screening.

Probiotic and lactulose: influence on gastrointestinal flora and pH value in minimal hepatic encephalopathy rats.

AIM: The present study was conducted to investigate the influence on gastrointestinal flora, counts of bifidobacteria and Enterobacterceae in colon and pH value of gastrointestinal after lactulose and probiotic treatment on rat experimental minimal hepatic encephalopathy (MHE) induced by thioactamide (TAA). METHODS: MHE was induced by intraperitoneal injection of TAA. 48 male MHE models were then randomly divided into 4 groups: control group (n = 12); MHE group (n = 12) received tap water ad libitum only; lactulose group (n = 12) and probiotics group (n = 12) gavaged respectively with 8 ml/kg of lactulose and 1.5 g/kg of probiotic preparation Golden Bifid (highly concentrated combination probiotic) dissolved in 2 ml of normal saline, once a day for 8 days. The latency of Brainstem auditory evoked potentials (BAEP) I was used as objective index of MHE. Counts of gastrointestinal flora, counts of bifidobacteria and Enterobacterceae in colon and pH value of gastrointestinal were examined respectively. RESULTS: Compared to MHE group, counts of gastrointestinal flora has greatly decreased, ratio of bifidobacteria and Enterobacterceae has greatly increased, pH value of colon has greatly descended (P < 0.05). However, there was no significant difference between lactulose group and probiotic group (P > 0.05). Both lactulose and probiotics can effectively prevent bacteria translocation and overgrowth, intensify CR, improved value of B/E, and acidify intestinal, decreased pH value of colon. CONCLUSION: Probiotic compound Golden Bifid is as useful as lactulose for the prevention and treatment of MHE. Probiotic therapy may be a safe, natural, well-tolerated therapy appropriate for the long-term treatment of MHE.

Molecular mechanisms of myocardial nitroso-redox imbalance during on-pump cardiac surgery.

BACKGROUND: The mechanism responsible for left ventricular dysfunction after cardiac surgery is only partly understood. In isolated rat hearts subjected to an ischaemia-reperfusion protocol, left ventricular dysfunction was associated with uncoupling of endothelial nitric oxide synthase (NOS) activity secondary to oxidation of the NOS cofactor, tetrahydrobiopterin (BH4). Here we investigated the effect of cardiopulmonary bypass and reperfusion on myocardial nitroso-redox balance in patients undergoing cardiac surgery. METHODS: From 116 patients who underwent elective cardiac surgery on cardiopulmonary bypass, paired samples of the right atrial appendages were obtained before venous cannulation of the right atrium and after myocardial reperfusion. Superoxide production from atrial samples was measured by lucigenin (5 µmol/L) enhanced chemiluminescence and 2-hydroxyethidium (2-OHE) detection by high-performance liquid chromatography (HPLC). BH4, oxidised biopterins, GTP-cyclohydrolase 1 (GTPCH-1, the rate-limiting enzyme in BH4 synthesis), and NOS activity ((14)C L-arginine to L-citrulline conversion) were measured by HPLC. FINDINGS: Atrial superoxide production increased significantly after reperfusion (from mean 37·83 relative light units per s per mg [SE 3·71] before cannulation to 65·02 [6·01] after reperfusion, p<0·0001; n=46 samples from 23 patients) due to increased mitochondrial and NOX2 oxidase activity (by 309% and 149%; p=0·002 and p=0·0002, respectively) and uncoupling of NOS activity. Atrial content of BH4 after perfusion was reduced (by 32%, p=0·001), as was activity of GTPCH1 (50%, p<0·0001). NOS activity decreased significantly after reperfusion (60%, p=0·0005) and this reduction was not affected by BH4 supplementation (10 µM) or NOX2 inhibition ex vivo. Instead, we identified increased endothelial NOS s-glutathionylation as the main mechanism for NOS uncoupling after reperfusion. Reversing NOS s-glutathionylation with dithiothreitol (100 µmol/L) completely restored NOS activity after reperfusion (p=0·34). INTERPRETATION: Our findings suggest that NOS s-glutathionylation, rather than BH4 depletion, accounts for NOS dysfunction in patients after cardiac surgery and cardiopulmonary bypass. FUNDING: British Heart Foundation.

Scrotal heat stress causes sperm chromatin damage and cysteinyl aspartate-spicific proteinases 3 changes in fertile men.

PURPOSE: To observe changes in semen parameters, sperm DNA integrity, chromatin condensation and cysteinyl aspartate-spicific proteinases (Caspase-3) in adult healthy men after scrotal heat stress (SHS). METHODS: The scrotums of 19 healthy male volunteers were exposed to the condition of 40-43 °C SHS belt warming 40 min each day for successive 2 days per week. The course of SHS was continuously 3 months. Routine semen analysis, hypo-osmotic swelling (HOS) test, eosin Y (EY) staining sperm HOS and chromatin dispersion (HOS/SCD) test, HOS and aniline blue (HOS/AB) staining test were carried out before, during and after SHS. The activated Caspase 3 levels of spermatozoa were determined with a microtiter plate reader. RESULTS: The mean parameters of sperm concentration, motility and normal morphological sperm were significantly decreased in groups with sperm being collected during SHS 1, 2 and 3 months when compared with those in groups of pre-SHS (P < 0.01). Statistically significant differences of sperm DNA fragmentation, normal sperm membrane and vitality, and Caspase-3 activity were observed between the groups of before SHS and after SHS 3 months and the groups of during SHS 1, 2 and 3 months (P < 0.001). Three months the SHS stopped, various parameters recovered to the level before SHS. Abnormal sperm with HOS/AB and HOS/SCD showed a negatively significant correlation with normal sperm by HOS/EY test, and WBC in semen showed a positively significant correlation with Caspase-3 activity. The percentage of abnormal sperm by using the test of HOS/SCD showed a positively significant correlation with that of HOS/AB. CONCLUSIONS: The continuously constant SHS can impact the semen quality, sperm DNA integrity, chromatin condensation and Caspase-3, and the combination of HOS plus AB test may simultaneously determine the integrity of membrane and chromatin condensation at the same spermatozoon.