Establishment of a cell line from the kidney of black carp and its susceptibility to spring viremia of carp virus.

In this study, we established and characterized a cell line derived from the kidney of black carp (Mylopharyngodon piceus), which is an important freshwater aquaculture species. The cell line was designated as MPK and subcultured for more than 70 passages in DMEM medium containing 10% fetal bovine serum (FBS) at 28°C. MPK had a modal diploid chromosome number of 48. Moreover, a transient MPK transfection efficiency was up to 18% using a green fluorescent protein plasmid by a modified electroporation. In addition, the MPK cells showed susceptibility to spring viremia of carp virus (SVCV), as demonstrated by the presence of severe cytopathic effects (CPEs) and increased viral RNA. Unexpectedly, the MPK cells expressed pluripotency-associated genes such as nanog, oct4 and vasa, indicating that these are possibly adult stem cells. Taken together, we have established a stable cell line from kidney that may potentially be utilized as an in vitro platform for genetic modifications and host-pathogen analysis in black carp.

Scaling the dynamic electron scattering in imaging the graphene sheets by the high-angle annular dark-field microscopy.

Employing the graphene sheets (GSs), the electron scattering constants are measured in the high-angle annular dark-field (HAADF) imaging by the scanning transmission electron microscopy. Single scattering is found to be dominant until the layer number of 200, complying with a simple relation of I = Io(1 - e(-tau/lambda)). The discrete layer counting of the GSs enables precise determination of incident depths. This work results values of lambda = 48.2, 61.4, 97.9 and 115.6 nm for 80, 120, 160 and 200 keV electrons, respectively. The uncertainties with the mean free paths and the cross sections are confined to 10 percent. The dependences on the electron beam energy and the collection angle are discussed based on a multislice simulation.

[Effects of vestibular spontaneous nystagmus on visual smooth pursuit function].

Objective: The aim of the study is to analyze the effects of vestibular spontaneous nystagmus(SN) on the smooth pursuit function of visual ocularmotor system. Methods: A total of 46 patients with acute unilateral peripheral vestibular syndrome with SN (26 cases of vestibular neuritis, 6 cases of Ramsay Hunt Syndrome (RHS) with vertigo, 14 cases of sudden deafness with vertigo) were included in this work. In the study group, the results of SPT and SN test with videonystagmography(VNG) were also reviewed. Taking SPT parameters, the influence of SN intensity on SPT gain, asymmetry and waveform and their correlation were analyzed.SPSS19.0 software was used for statistical analysis. Results: Among the 46 patients, there were 36 cases of SN pointing to the healthy side(SN intensity range of 2.68°/s-32.53°/s), and 10 cases of SN pointing to the affected side (SN intensity range of 2.66°/s-16.54°/s). SN intensity was divided into 3 groups, including light(0.50°/s-5.00°/s), medium(5.01°/s-10.00°/s) and strong(>10.01°/s), accounting for 14 cases(30.4%), 18 cases(39.1%) and 14 cases(30.4%), respectively. The differences of the gain of SPT to the fast phase and slow phase direction in the overall groups and light, medium and strong groups of SN intensity respectively were statistically significant(ttotal=13.338, tlight=6.184, tmedium=8.436, tstrong=8.477, all of P<0.001). The difference of SPT gain in SN fast phase direction between groups with different SN intensity was statistically significant(F=9.639, P<0.001),there was no statistically significant difference in SPT gain between the groups on the SN slow phase direction(F=1.137, P=0.330).The SN intensity significantly negatively correlated with the SPT gain of the fast phase direction of SN (r=-0.433, P=0.003), that was, the SPT gain on the fast phase direction of SN decreased with the increase of SN intensity. There was no significant correlation between SN intensity and the gain of SPT on the slow phase direction of SN (r=-0.061, P=0.687). SPT waveform analysis showed that type I, type II and type III accounted for 8 cases(17.4%), 21 cases(45.6%) and 17 cases(37.0%), respectively. The corresponding mean values of SN intensity were (3.71±0.69)°/s, (7.44±1.88)°/s, (20.04±5.53)°/s, respectively, without type IV wave. The intensity of SN was positively correlated with the asymmetric value of the gain of SPT left and right(r=0.450,P=0.002). That was, with the increase of SN strength, the asymmetric value also increased, and the worse the asymmetry of the gain of SPT left and right pursuit was, the worse the SPT waveform was. Conclusion: SPT gain, asymmetry and SPT waveforms are all affected by SN, and the greater the intensity of SN, the greater the influence on the three. When SN is strong, type III waves may occur, suggesting that acute peripheral vestibular syndrome can also affect the visual ocularmotor systems.

[Analysis of broadband sinusoidal harmonic acceleration test results for normal young people].

Objective: We tested more frequency (0.01-3.0 Hz) of the sinusoidal harmonic acceleration test(SHAT) by the NKI rotation chair, aimed to analyze the results of the normal youth. Methods: Fifty normal young people were tested with sinusoidal harmonic acceleration by NKI rotation chair. Including 24 men and 26 women, aged from 20 to 40 years old, with median age of 27 years, with no vestibular dysfunction participated in the present study. All the volunteers accepted the sinusoidal harmonic acceleration test by the NKI rotation chair, the frequency included 0.01 Hz, 0.02 Hz, 0.03 Hz, 0.05 Hz, 0.1 Hz, 0.2 Hz, 0.3 Hz, 0.6 Hz, 1.0 Hz, 1.5 Hz, 2.0 Hz, 3.0 Hz. Taken the gain, phase, and asymmetry as the parameters, the trait of each frequency was analysed. Results: From 0.01 Hz to 1.5 Hz, the gain increased close to 1 gradually, and with fluctuates slightly at the 2.0 Hz and 3.0 Hz frequencies. The phase decreased as frequencies increased close to 0 at 0.3 Hz. The frequency above 0.6 Hz presented negative. Asymmetric values were distributed on both positive and negative sides at each frequency and were different from each other, both values were less than 20%. Conclusion: Broadband SHAT can accurately assess the horizontal semicircular canal function at different frequencies in a wider frequency range, and objectively, reflect the physiological function of the horizontal semicircular canal, and expand the clinical value of traditional SHAT.

Orally administered resveratrol enhances the therapeutic effect of autologous transplanted adipose-derived stem cells on rats with diabetic hepatopathy.

Stem cell therapy is a promising treatment for hepatopathy due to diabetes mellitus (DM); oral resveratrol treatment exhibits protective effects. We investigated whether protective effects could be produced in liver of diabetic rats receiving autologous adipose-derived stem cell transplantation (ADSC) plus oral resveratrol administration. Male rats were divided into four groups: sham group; streptozotocin induced DM group; DM + ADSC group, in which DM rats were treated with 106 stem cells/rat; and DM + R + ADSC group, in which DM rats were treated with ADSC and oral resveratrol. The DM group exhibited apoptosis, inflammation and fibrosis, whereas Sirt-1 and survival signaling were suppressed. Pathological conditions other than survival signaling were improved in the DM + ADSC group. All pathological conditions were improved in the DM + R + ADSC group. Also, the oxidative stress level in the blood was reduced in the DM + R + ADSC group compared to the sham group. Oral resveratrol administration appears to reduce oxidative damage and enhances survival signaling in diabetic liver. The therapeutic response in the DM + R + ADSC group was better than in the DM + ADSC group.

[The characteristics of semicircular canal injury frequency in vestibular neuritis patients].

Objective:To evaluate the frequency characteristics of semicircular canal injury in patients with vestibular neuritis. Method:Fifty-three patients with vestibular neuritis were respectively evaluated by caloric test(CT)，head shaking test(HST),video head impulse test (vHIT),which reflects semicircular canal function in the low, middle, high frequency region. The asymmetric value of CT unilateral semicircular canal reaction (UW), head shaking nystagmus induced by HST, gain value of VHIT (VHIT-G) and presence or absence of saccade (VHIT-S) were used as observation indicators. The data were analyzed by SPSS 17.0 statistical software.The characteristics of the results of the three tests in patients with vestibular neuritis was analyzed and the functional status of the high, middle and low frequency regions of the semicircular canal was evaluated. Result:The positive rates of CT, HST, VHIT-G and VHIT-S in 53 patients with vestibular neuritis were 94.3%（50/53）,75.5%（40/53）,81.1%（43/53）,96.2%（51/53）,respectively. The positive rates of CT, HST, VHIT-G and VHIT-S were 92.9%（26/28）,89.3%（25/28）,92.9%（26/28）,96.4%（27/28）in 28 cases with disease course ≤ 7 days, and the positive rates in 25 patients with disease course >7 days were 96.0%（24/25）,60.0%（15/25）,68.0%（17/25）,96.0%（24/25）,respectively. Compared with the positive rate of each index, CT and VHIT-G (P=0.076), HST and VHIT-G (P=0.480) had no statistical significance. The difference between CT and HST (P=0.015), VHIT-G and VHIT-S(P=0.032) was statistically significant. The positive rates of CT, HST, VHIT-G and VHIT-saccade in patients with course of disease ≤7 days were compared, and there was no significant difference. In patients with disease course >7 days, except HST and VHIT-G, there were statistical differences in other indexes. The correlation test between VHIT-G and VHIT-S：r=-0.437, P=0.006. Conclusion:The vestibular injury in patients with vestibular neuritis showed in a manner of full frequency injury. The multi-frequency detection technique is helpful for the clinical diagnosis of vestibular neuritis, and also can reflect the compensation and recovery of vestibular function.

[Objective characteristics of nystagmus in patients with posterior semicircular canal benign paroxysmal positional vertigo].

Objective: To analyze and discuss the parameters and clinical significance of nystagmus in patients with benign positional paroxysmal vertigo (BPPV) of posterior semicircular canal. Methods: The subjects of the study were 564 BPPV patients diagnosed with posterior semicircular canal canalithis (PSC-can) from January 2016 to July 2017 in Tianjin No.1 Central Hospital, including 186 males and 378 females, with a median age of 57 years. The induced nystagmus in Dix-Hallpike test was recorded by video nystagmuo graph(VNG), and the direction, latency, duration time and intensity characteristics of nystagmus were compared with the position of hanging and sitting.SPSS17.0 software was used for statistical analysis. Results: Vertical torsional nystagmus was both induced with the position of hanging and sitting during Dix-Hallpike test. The vertical direction of the induced nystagmus was upward and downward respectively. The latency, duration time and intensity of lesion side were L(2.65±1.92; 1.44±1.24), D(14.90±10.46; 15.28±8.06), and P(29.75±21.26; 14.08±9.48). The latency and intensity in hanging position were higher than those of sitting. The intensity rate was about 2∶1, with statistically significant difference (t=13.831, and 17.296, P all<0.01). The direction of the induced nystagmus was opposite to turning in HSC-Can BPPV. The intensity turning to normal side was larger than lesion side obviously. The intensity rate was about 2∶1, the difference was statistically significant (t=17.296, P<0.01). There was no statistical difference of nystagmus during time between the two positions(t=-0.735, P>0.05). Conclusions: The nystagmus intensity rate in Dix-Hallpike test between hanging and sitting position of lesion side in PSC-Can is 2∶1, which conforms to the Ewald's law. The direction, latency and intensity of nystagmus can be used as a reference index for the localization diagnosis of PSC-Canotolith.

[Preliminary effect observation on the application of micro-negative pressure in children with small-area deep partial-thickness burn].

Objective: To preliminarily observe the effects of application of micro-negative pressure in children with small-area deep partial-thickness burn. Methods: From January 2016 to August 2018, 64 children with small-area deep partial-thickness burn who were admitted to the Department of Burn Surgery of the First Affiliated Hospital of Naval Medical University were recruited in this prospective randomized controlled study. According to the random number table, they were divided into negative pressure group [18 boys and 14 girls, aged (3.9±1.6) years with total burn area of (5.5±2.2)% total body surface area (TBSA)] and conventional group [20 boys and 12 girls, aged (3.8±1.7) years with total burn area of (5.8±1.6)% TBSA], with 32 patients in each group. After admission, simple debridement was performed in the patients of 2 groups. After that, the children in negative pressure group were treated with micro-negative pressure with negative pressure material replaced every 3 to 5 days. Children in conventional group were treated with silver sulfadiazine cream with dressing change every other day. On post injury day (PID) 14 and 21, general wound observation was performed, the wound healing rate was calculated, the exudates from the wounds were cultured and the positive detection rate was calculated. The number of patients requiring surgical skin grafting was recorded and the rate of surgical skin grafting was calculated, and the complete wound healing time was recorded in the patients of 2 groups. Scar formation was evaluated by the Vancouver Scar Scale (VSS) in 3, 6, and 12 months after wound healing. Data were processed with chi-square test, t test, Bonferroni correction, and analysis of variance for repeated measurement. Results: (1) On PID 14, all the necrotic tissue in the wounds of patients in negative pressure group was removed, with few exudates, and most of the wounds had been epithelialized; most of necrotic tissue in the wounds of patients in conventional group was removed, with more exudates and smaller wound healing area than those in negative pressure group. On PID 21, most of the wounds of patients in negative pressure group were healed, and the exudates were rare, while the wound healing area of patients in conventional group was significantly smaller than that in negative pressure group with more exudates. (2) On PID 14 and 21, the wound healing rates [(49.8±3.3)% and (95.8±2.4)%] of patients in negative pressure group were significantly higher than those in conventional group [(40.0±3.2)% and (75.3±2.5)%, t=11.899, 33.461, P<0.01]. (3) On PID 14 and 21, the positive detection rates of wound bacteria of patients in negative pressure group were significantly lower than those in conventional group (χ(2)=6.275, 5.741, P<0.05). (4) The rate of surgical skin grafting of patients in negative pressure group was significantly lower than that in conventional group (χ(2)=5.333, P<0.05). (5) The complete wound healing time of patients in negative pressure group [(23.9±2.3) d] was significantly shorter than that in conventional group [(27.9±1.8) d, t=-7.806, P<0.01]. (6) In 3, 6, and 12 months after wound healing, the VSS scores [(6.9±1.8), (5.6±1.4), (3.4±1.5) points] of patients in negative pressure group were significantly lower than those in conventional group [(9.0±1.5), (7.4±2.0), (5.7±1.6) points, t=-4.987, -4.127, -5.988, P<0.01]. Conclusions: In comparison with routine dressing change, the treatment of application of micro-negative pressure in children with small-area deep partial-thickness burn can significantly improve the wound healing rate and rate of surgical skin grafting, decrease the wound infection rate, shorten the wound healing time, and improve the wound healing quality.

[The nystagmus characteristics of two positions of the horizontal semicircular canal cupulolithiasis].

Objective: To analyze the characteristics of nystagmus of horizontal semicircular canal cupulolithiasis(HSC-Cup) in Roll test and Dix-Hallpike test. Methods: Between December 2016 and December 2017, a total of 164 patients with BPPV from Tianjin First Center Hospital, 124 HSC-Can BPPV and 40 HSC-Cup BPPV, were involved.The induced nystagmus in Roll test and Dix-Hallpike test were recorded by video-nystagmograph(VNG), whose direction and intensity characteristics were compared in various BPPV. Results: HSC-Can patients were induced a horizontal nystagmus with Roll test, the nystagmus intensity of the disease and healthy side were (41.3±20.1)°/s (mean standard deviation)and(21.9±9.4)°/s respectively, the difference was statistically significant (t=6.709, P<0.05). HSC-Cup patients were induced a horizontal nystagmus that was opposite to the direction of the Roll test, the nystagmus intensity of the disease and the healthy side were (12.9±6.4)°/s and(29.1±9.3)°/s respectively, with significant difference (t=9.066, P<0.05). Among 124 cases of HSC-Can patients, 120 cases of horizontal nystagmus were recorded in the left and right sides of the Dix-Hallpike test in the same direction as the turning direction, the horizontal nystagmus intensity of the disease and the healthy sides were (15.1±10.0)°/s and(9.4±7.4)°/s respectively, the difference was statistically significant (t=2.365, P<0.05). And 40 cases of HSC-Cup patients were recorded at the head of the Dix-Hallpike test in the opposite direction to the horizontal eye earthquake.The horizontal nystagmus intensity of the disease and the healthy side were (5.3±2.8)°/s and(13.9±4.4)°/s respectively, the difference was statistically significant (t=10.579, P<0.05). Conclusions: Characteristic horizontal nystagmus can be induced by HSC-Cup in both roll test and Dix-Hallpike test. Preliminary localization of Dix-Hallpike Test can also be used for HSC-Cup.

[The characteristics and clinical significance of vestibular autorotation test in patients with vestibular migraine].

Objective: To investigate the characteristics and clinical utility of vestibular autorotation test (VAT) in patients with vestibular migraine(VM). Methods: This study included two groups, a VM group (441 patients from Tianjin First Center Hospital between January 2015 and May 2016) and a control group (65 healthy subjects). Both groups undertook VAT; the parameters evaluated were horizontal gain/phase, vertical gain/phase and asymmetry. The differences in VAT results between the two groups were investigated. Results: There were statistically significant differences in VAT results between the VM and the control group, namely elevated horizontal gain at frequency 2, 3, 4 and 5 Hz, delay horizontal phase at frequency 2, 4, 5 and 6 Hz, elevated vertical gain at frequency 2-6 Hz and delay vertical phase at frequency 4-6 Hz. There was no significant difference in asymmetric values between the VM group and the control group. Conclusions: The results of this study indicate that VM patients have elevated horizontal gain and vertical gain, and delay horizontal phase and vertical phase. It is suggested that VAT represents a useful diagnostic tool which may provide objective evidence for the diagnosis and differential diagnosis of VM.

[A pilot study of the unilateral centrifugation subjective visual vertical in healthy young people].

Objective: To evaluate the data of unilateral centrifugation subjective visual vertical (UC-SVV) in healthy young people.To study the function of utricle. Methods: Between Decem ber 2017 and May 2018, thirty-two healthy young volunteers were tested by static subjective visual vertical(SVV) and low velocity UC-SVV with Neuro Kinetics Inc I-portal 6.0 Video nystagmus recording system and NOTC rotating chair system.The static SVV preset angle were -15.00°, 15.00°, -20.00°, 20.00°, -12.00° and 12.00° respectively.UC-SVV test parameters: rotating chair's peak speed was 60°/s.The shift time from the middle to the lateral position was 30 s and the displacement was 3.85 cm.The chair rotated at 60 s at left, right and middle positions.Subjects underwent SVV during this period.SPSS 17.0 software was used to analyze the data. Results: The mean and standard deviation of static SVV deviation in 32 volunteers was 0.21°±0.17°, 95%CI (-0.14°, 0.55°). The mean and standard deviation of UC-SVV in the left, middle and right positions of the clockwise was 0.24°±0.25°, -0.10°±0.27°, -0.63°±0.26°, 95%CI(-0.26°, 0.74°), (-0.65°, 0.44°), (-1.16°, -0.10°). The mean and standard deviation of UC-SVV in the left, middle and right positions of the counter clockwise was 0.03°±0.27°, -0.11°±0.26°, -0.23°±0.26°, 95%CI(-0.52°, 0.59°), (-0.65°, 0.42°), (-0.76°, 0.30°). There was significant difference in the deviation between the left and the right side of counter clockwise rotation of UC-SVV(t=2.432, P<0.05), however, there was no significant difference in the angle of deviation between the left and right sides and the median position(t value was 0.951, -1.400, both P>0.05). There was no significant difference among each position in clockwise rotation of UC-SVV(F=0.253, P>0.05). Conclusion: UC-SVV test with peak speed of 60°/s can cause vertical line deviation in the left and right lateral position, but the angle is small and the value of clinical application is limited.

[Debridement combined with vacuum sealing drainage in the treatment of severe infection in abdominal wall due to allogeneic umbilical cord embedded in abdominal wall for immunotherapy].

Objective: To explore the effect of debridement combined with vacuum sealing drainage (VSD) on the treatment of severe infection in abdominal wall due to allogeneic umbilical cord embedded in abdominal wall for immunotherapy. Methods: From January 2015 to December 2016, 12 patients with severe infection in abdominal wall due to allogeneic umbilical cord embedded in abdominal wall for immunotherapy were admitted to our department. They were conducted with systemic anti-infective treatment, local debridement, and VSD. The wounds were continuously washed for 3 to 5 days after the VSD device installed, with negative pressure value from -16.0 to -12.0 kPa. The VSD device was removed 5 to 7 days later. Continue wound dressing by aseptic ribbon gauze was stuffed in the cavity, and the incision was sutured after the granulation tissue grew well in the cavity. Results: In all patients, allogeneic umbilical cords were completely removed and abdominal infection was cured. The wounds healed well, the sensory function of abdominal was normal, and the activity was not restricted. All the patients were followed up for 3 to 6 months with no reinfection or incisional hernia. Conclusions: Embeding the whole allogeneic umbilical cord in abdominal wall for immunotherapy can lead to severe infection in abdominal wall. Abdominal infection can be cured by debridement combined with VSD with good clinical results.

C-terminus of Hsc70-interacting protein (CHIP) enhances stemness properties of human Wharton's jelly mesenchymal stem cell.

Mesenchymal stem cells are an attractive source of multipotent cells in part because they are easy to obtain. Several E3 ligases regulate the stability and functions of various factors in different adult stem cells through the ubiquitylation pathway. We investigated the C-terminus of Hsc70-interacting protein (CHIP) E3 ligase that regulates pluripotency of human Wharton's jelly mesenchymal stem cells (hWJMSC). We found that CHIP increases protein kinase B (Akt) phosphorylation by decreased expression of phosphatase and tensin homolog (PTEN), which suggests improvement of the survival pathway by CHIP over-expression. We also found that increased CHIP expression induced Sox2 and NANOG, which can promote stem cell self-renewal and prevent oxidative stress-induced senescence of hWJMSC by decreased p21. We found that CHIP could be used to enhance the multiple functions of hWJMSC.

Epidemiology and outcomes of bloodstream infections in 177 severe burn patients from an industrial disaster: a multicentre retrospective study.

OBJECTIVES: To determine the characteristics of bloodstream infections (BSIs) and to evaluate the impact of BSIs on mortality in severe burn patients. METHODS: A retrospective observational study was conducted in 20 tertiary hospitals. A total of 185 patients who experienced a massive dust explosion in eastern China were included. RESULTS: After exclusion, 177 patients were analysed. The median total body surface area (TBSA) burned was 95% (interquartile range 85%-98%). Inhalation injuries occurred in 97.2%. The overall 90-day mortality was 35% (62/177). During the study period, 120 (67.8%) patients developed 253 episodes of BSI with 323 unique causative pathogens. Sixty-six episodes were polymicrobial infections. Catheter-related BSIs (CRBSIs) accounted for 41.5% of the episodes. Acinetobacter baumannii (19.5%), Klebsiella pneumoniae (13.9%) and Candida (12.7%) were the most common organisms. Antimicrobial resistance was found in 63.5% of the isolates, particularly in Gram-negative bacteria. Patients who developed BSIs had a greater illness severity at admission to the intensive care unit, and worse outcomes. After adjusting for demographics, severity of illness and treatment characteristics in a multivariate logistic model, there was a trend toward BSI increasing the risk of 90-day mortality (adjusted OR 3.4; 95% CI 0.9-12.9; p=0.069). In subgroup analyses, CRBSIs (adjusted OR 5.7; 95% CI 1.3-24.9; p=0.021 versus no BSI) and polymicrobial BSIs (adjusted OR 6.1; 95% CI 1.3-28.1; p=0.020 versus no BSI) had greater risk of 90-day mortality. CONCLUSIONS: A strikingly high rate of BSIs was observed in severe burn patients. Gram-negative organisms and fungi were the leading causes. CRBSIs and polymicrobial BSIs were associated with high mortality.

Reproducing abnormal cholesterol biosynthesis as seen in the Smith-Lemli-Opitz syndrome by inhibiting the conversion of 7-dehydrocholesterol to cholesterol in rats.

The Smith-Lemli-Opitz syndrome is a recessive inherited disorder characterized by neurologic developmental defects and dysmorphic features in many organs. Recently, abnormal cholesterol biosynthesis with impaired conversion of 7-dehydrocholesterol to cholesterol has been discovered in homozygotes. To reproduce the biochemical abnormality, BM 15.766, a competitive inhibitor of 7-dehydrocholesterol-delta 7-reductase, the enzyme that catalyzes the conversion of 7-dehydrocholesterol into cholesterol was fed by gavage to rats. After 14 d, plasma cholesterol concentrations declined from 48 mg/dl to 16 mg/dl and 7-dehydro-cholesterol levels rose from trace to 17 mg/dl. Hepatocytes surrounding the central vein developed balloon necrosis. Stimulating cholesterol synthesis with cholestyramine followed by BM 15.766 produced an additional 40% decline (P < 0.05) in plasma cholesterol and 34% increase in 7-dehydrocholesterol levels compared to the inhibitor alone. Adding 2% cholesterol to the diet during the second week of BM 15.766 treatment increased plasma cholesterol threefold and decreased 7-dehydrocholesterol concentrations 55%. Hepatic 3-hydroxy-3-methylglutaryl co-enzyme A (HMG-CoA) reductase activity increased 73% with a 3.9-fold rise in mRNA levels but cholesterol 7 alpha-hydroxylase activity decreased slightly though mRNA levels increased 1.4 times with BM 15.766 treatment. These results demonstrate that BM 15.766 is a potent inhibitor of 7-dehydrocholesterol-delta 7-reductase. The model reproduces abnormal cholesterol biosynthesis as seen in the Smith-Lemli-Opitz syndrome and is useful to test different treatment strategies. Stimulating early steps of cholesterol synthesis worsens the biochemical abnormalities while feeding cholesterol inhibits abnormal synthesis, improves the biochemical abnormalities and prevents liver damage.

Increasing dietary cholesterol induces different regulation of classic and alternative bile acid synthesis.

We investigated the effect of increasing dietary cholesterol on bile acid pool sizes and the regulation of the two bile acid synthetic pathways (classic, via cholesterol 7alpha-hydroxylase, and alternative, via sterol 27-hydroxylase) in New Zealand white rabbits fed 3 g cholesterol/per day for up to 15 days. Feeding cholesterol for one day increased hepatic cholesterol 75% and cholesterol 7alpha-hydroxylase activity 1.6 times without significant change of bile acid pool size or sterol 27-hydroxylase activity. After three days of cholesterol feeding, the bile acid pool size increased 83% (P < 0.01), and further feeding produced 10%-20% increments, whereas cholesterol 7alpha-hydroxylase activity declined progressively to 60% below baseline. In contrast, sterol 27-hydroxylase activity rose 58% after three days of cholesterol feeding and remained elevated with continued intake. Bile drainage depleted the bile acid pool and stimulated downregulated cholesterol 7alpha-hydroxylase activity but did not affect sterol 27-hydroxylase activity. Thus, increasing hepatic cholesterol does not directly inhibit cholesterol 7alpha-hydroxylase and initially favors enzyme induction, whereas increased bile acid pool is the most powerful inhibitor of cholesterol 7alpha-hydroxylase. Sterol 27-hydroxylase is insensitive to the bile acid flux but is upregulated by increasing hepatic cholesterol.

Unexpected inhibition of cholesterol 7 alpha-hydroxylase by cholesterol in New Zealand white and Watanabe heritable hyperlipidemic rabbits.

We investigated the effect of cholesterol feeding on plasma cholesterol concentrations, hepatic activities and mRNA levels of HMG-CoA reductase and cholesterol 7 alpha-hydroxylase and hepatic LDL receptor function and mRNA levels in 23 New Zealand White (NZW) and 17 Watanabe heritable hyperlipidemic (WHHL) rabbits. Plasma cholesterol concentrations were 9.9 times greater in WHHL than NZW rabbits and rose significantly in both groups when cholesterol was fed. Baseline liver cholesterol levels were 50% higher but rose only 26% in WHHL as compared with 3.6-fold increase with the cholesterol diet in NZW rabbits. In both rabbit groups, hepatic total HMG-CoA reductase activity was similar and declined > 60% without changing enzyme mRNA levels after cholesterol was fed. In NZW rabbits, cholesterol feeding inhibited LDL receptor function but not mRNA levels. As expected, receptor-mediated LDL binding was reduced in WHHL rabbits. Hepatic cholesterol 7 alpha-hydroxylase activity and mRNA levels were 2.8 and 10.4 times greater in NZW than WHHL rabbits. Unexpectedly, cholesterol 7 alpha-hydroxylase activity was reduced 53% and mRNA levels were reduced 79% in NZW rabbits with 2% cholesterol feeding. These results demonstrate that WHHL as compared with NZW rabbits have markedly elevated plasma and higher liver cholesterol concentrations, less hepatic LDL receptor function, and very low hepatic cholesterol 7 alpha-hydroxylase activity and mRNA levels. Feeding cholesterol to NZW rabbits increased plasma and hepatic concentrations greatly, inhibited LDL receptor-mediated binding, and unexpectedly suppressed cholesterol 7 alpha-hydroxylase activity and mRNA to minimum levels similar to WHHL rabbits. Dietary cholesterol accumulates in the plasma of NZW rabbits, and WHHL rabbits are hypercholesterolemic because reduced LDL receptor function is combined with decreased catabolism of cholesterol to bile acids.

[Advances in preparation and clinical application of amniotic membrane graft].

Amniotic membrane has been used as wound dressing for more than 100 years. With the development of the preservation and preparation techniques, amniotic membrane is widely used in ophthalmology, burns, plastic surgery, dentistry, and neurosurgery. In recent years, as more and more amniotic membrane is used in chronic wounds, it will be a new treatment method for wounds. This paper is a brief review about advances in preparation and clinical application of amniotic membrane graft.

[Effects of caspase-1 inhibitor VX765 on cold-restraint stress-induced acute gastric ulcer in mice].

Objective: To investigate the protective effects of caspase-1 inhibitor VX765 on gastric mucosa of mice with cold-restraint stress-induced acute gastric ulcer. Methods: Twenty-four specific pathogen free male C57BL/6J mice were divided into normal control group (NC), cold restrain group (CR), VX765 pre-treatment+ cold restrain group (VCR), and rabeprazole pre-treatment+ cold restrain group (RCR) according to the random number table, with 6 mice in each group. Mice in group NC were injected intraperitoneally with solution of 10 mL/kg dimethylsulfoxide (DMSO) and phosphate buffer solution (PBS). Mice in group CR were inflicted with acute gastric ulcer induced by cold-restraint stress 30 minutes after intraperitoneal injection of solution of DMSO and PBS. Mice in groups VCR and RCR were inflicted with acute gastric ulcer as above 30 minutes after intraperitoneal injection of solution of DMSO and PBS with dose of 12.5 µmol/kg containing 10 mg VX765 and 40 mg/kg containing 20 mg rabeprazole, respectively. Four hour after cold-restraint stress, serum content of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6)was determined by enzyme-linked immunosorbent assay. Gross condition of gastric tissue was observed. Ulcer index was evaluated. Pathological change of gastric tissue was observed with HE staining. The relative expression of IL-1ß, IL-18, and cleaved-caspase-1 in gastric tissue were detected by Western blotting. Mice in group NC were detected as above at the same time point. Data were processed with one-way analysis of variance and Bonferroni test. Results: The serum content of TNF-α and IL-6 and the relative expression of cleaved-caspase-1, IL-1ß, and IL-18 in gastric tissue of mice in group NC were significantly lower than those in group CR (with P values below 0.01). The content of the above-mentioned inflammatory indexes in serum and gastric tissue of mice in group VCR was significantly lower than that in group CR (with P values below 0.01). There were no statistically significant differences in content of the above-mentioned inflammatory indexes in serum and gastric tissue of mice between groups RCR and CR (with P values above 0.05). The content of the above-mentioned inflammatory indexes in serum and gastric tissue of mice in group VCR was significantly lower than that in group RCR (with P values below 0.01). Surface of gastric mucosa was smooth and morphology of mucosal cells was normal with clear structure of mice in group NC. Multiple hemorrhage of gastric mucosa, disorderly arrangement of mucosal cells, and large number of inflammatory cell infiltration around necrotic tissue were observed in mice of group CR. Decreased number of gastric mucosa bleeding, intact mucosal structure, and small amount of inflammatory cell infiltration around necrotic tissue were observed in mice of groups VCR and RCR. The ulcer indexes of mice in groups NC, CR, VCR, and RCR were 0, 18.7±1.1, 6.3±1.5, and 8.2±1.3, respectively. The ulcer index of mice in group NC was significantly lower than that in the other 3 groups (with P values below 0.05). The ulcer indexes of mice in groups VCR and RCR were close (P>0.05), which were significantly lower than ulcer index of mice in group CR (with P values below 0.05). Conclusions: VX765 can effectively inhibit the activation of caspase-1, reduce production of inflammatory factor, and alleviate inflammatory response, which have protective effects on gastric mucosa of mice with cold-restraint stress-induced acute gastric ulcer.

[The exploration on optimization of two alternatives between roll test and Dix-Hallpike test in benign paroxysmal positional vertigo].

Objective: To analyze the objective characteristics of roll test and Dix-Hallpike test in benign paroxysmal positional vertigo(BPPV)patients, discussing the premier solution of positional test. Methods: A total of 230 patients with BPPV, whereas 170 posterior semicircular canal canalithiasis (PSC-Can) BPPV and 60 horizontal semicircular canal canalithiasis (HSC-Can) BPPV were involved respectively. The induced nystagmus in roll test and Dix-Hallpike test was recorded by video nystagmuo graph (VNG), and the direction, intensity and time characteristics of nystagmus were compared in various BPPV.SPSS19.0 software was used for statistical analysis. Results: Vertically upward nystagmus was induced by hanging in 170 PSC-Can Dix-Hallpike test, and the nystagmus reversed and turned weaker when the subjects came to sit. The intensity of nystagmus at turning to lesion side by hanging and sitting were (30.3±14.1)°/s and (12.6±7.5)°/s respectively, the difference was statistically significant (t=20.153, P<0.05). However, no nystagmus was induced in PSC-Can roll test. Horizontal nystagmus in the same direction with turning was induced in 60 HSC-Can roll test. The intensity of nystagmus at turning to lesion side and normal side was (42.0±18.0)°/s and (20.3±8.7)°/s respectively, the difference was statistically significant (t=12.731, P<0.05). Furthermore, horizontal nystagmus in the same direction with turning was induced in 57 HSC-Can Dix-Hallpike. The coherence was 95% with the results of roll test. Conclusions: Dix-Hallpike test can not only be used to diagnose PSC-Can, but also induce nystagmus in HSC-Can effectively. Whereas the roll test only show significance in diagnosing HSC-Can. To avoid uncomfortable stimulation to patients as much as possible, we suggest to use Dix-Hallpike test at first, and to judge whether using roll test based on the result of the horizontal nystagmus.