Objective To observe morphological characteristics and activity distribution of T. asahii biofilm. Methods The morphological characteristics of T. asahii biofilm were observed under an inverted microscope and scanning electron microscope, and activity was measured and quantitatively analyzed by 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazo-lium hydroxide (XTT) assay and viable count, respectively. Spatial distribution of dead/vital cells, activity and thickness of biofilm at different layers were assessed under a confocal laser scanning microscope (CLSM) following double staining with FDA/PI. Results T. asahii formed a biofilm in vitro on the surface of polystyrene materials. Under a scanning microscope, the biofilm displayed a complex three-dimensional structure which composed of spores, pseudohy-pha and true hypha. As time prolonged, the activity and quantity of biofilm increased. The results of XTT assay were correlated with those of viable count (r = 0.94, P < 0.01). The activity was of no obvious difference between different layers of the biofilm. The thickness of biofilm varied from 14.3 μm to 31 μm. Conclusions The structure of T. asahii biofilm in vitro is more complex than that of planktonic T. asahii. The activity is of no significant difference between different layers of T. asahii biofilm.

Objective:To determine the content of self-manufactured and imported lurasidone hydrochloride tablets in order to e-valuate their internal qualities. Methods:The determination of lurasidone hydrochloride tablets was performed by HPLC. The HPLC system consisted of a Waters C8 column (250 mm × 4. 6 mm, 3. 5 μm) and the mobile phase of 0. 05 mol·L-1 phosphate buffer solu-tion (pH 3. 0)-acetonitrile(60∶40), the detection wavelength was 230 nm, the flow rate was 1. 2 ml·min-1 and the column tempera-ture was 40℃, and the injection volume was 20μl. Results:The linear range of lurasidone hydrochloride was 0. 100 8-0. 806 4 mg· ml-1(r=0. 999 5). The average recovery was 99. 95% with RSD of 0. 31%(n=9). Conclusion:The method is simple, rapid, ac-curate, and reliable. The method can determine lurasidone hydrochloride tablets satisfactorily. According to the results, there are few differences among the self-manufactured and imported lurasidone hydrochloride tablets.

Objective To discuss the methods of the heat preservation performance monitoring of the blood transport case and to provide the technical support for the safety of blood transportation.Methods At the different environment temperature,the amount of the cold resource was decided by the mass ratio of cold resource to blood and the temperature was automatically recorded by the intelligent temperature chip continuously,to monitor the changes of each monitoring point in the blood transport case.Results When the mass ratio of cold resource to blood was fixed at 1∶6,the cold chain of the blood transport case could keep the tempera-ture of 2-10 ℃ for 8 hours at the environment temperature of 12 ℃,It could keep the temperature of 2-10 ℃ for 4.5 hours at the environment temperature of 25 ℃,and it could keep the temperature of 2-10 ℃ for 2 hours at the environment temperature of 44℃.Conclusion When the mass ratio of cold resource to blood is fixed,as the environment temperature changes,the available time that the blood transport case keeps with the cold-chain requirement varies according to the results of the heat preservation per-formance monitoring of the blood transport case.

Objective:To investigate the effects of thermobaric charge explosion simulated gas on long-term neurobehavior and hippocampal neurogenesis in rats.Methods:A total of 48 male SPF grade SD rats aged 8-10 weeks were randomly divided into control group, 5 min exposure group, 10 min exposure group and 15 min exposure group, with 12 rats in each group. Twenty-eight days after inhalation of infection, the anxiety-like behavior of rats was evaluated by an elevated cross maze, and the learning and memory function of rats was evaluated by two-way active avoidance experiment. The number of positive cells of rat hippocampal dentate gyrus neural stem cells marker molecule neural epithelial cell protein (SOX2) and mature neuron marker molecular neuronal nuclei (NeuN) was detected by immunofluorescence staining. Western blot was used to detect SOX2 and NeuN protein expression in the hippocampal tissues of rats. GraphPad prism 8.0 software was used for data analysis.The comparison of repeated measurement design data was carried out by repeated measurement ANOVA.One-way ANOVA was used for inter group comparisons, and Tukey test was used for pairwise comparison. Hippocampal nerve cells were counted using the Image J software.Results:(1) The experimental results of the elevated cross maze showed that the percentage of arm opening and the percentage of open arm residence time in each group had significant group effects ( F=22.31, 5.43, all P<0.05). The percentage of open arm entry times of rats in the 5 min, 10 min and 15 min exposure group ((28.85±1.47)%, (15.04±4.69)%, (12.66±2.89)%) and the percentage of residence time in open arm ((12.12±2.64)%, (12.16±1.11)%, (8.73±3.52)%) were all lower than those of the control group ((65.40±1.86)%, (42.92±3.12)%) (all P<0.05). There were no statistically significant differences in pairwise comparison among the three exposure groups (all P>0.05). (2)During the memory acquisition period, the results of repeated-ANOVA showed that the time main effect ( F=56.46), the group main effect ( F=16.64) and the interaction effect had significant differences( F=4.21)(all P<0. 05). The difference values of active avoidance number between the 4th day and 1st day among the four groups were significant different ( F=68.63, P<0.05). During the memory reproduction period, there were significant differences in active avoidance number and active avoidance time among the four groups ( F=8.17, 8.28, both P<0.05). The active avoidance numbers in 10 min and 15 min exposure groups((2.50±0.26) times, (2.33±0.06) times)were significantly lower than those in the control group ((8.33±3.72) times) (both P<0.05), and the active avoidance time ((6.25±0.40)s, (6.61±1.63)s) were significantly higher than those in the control group((3.69±1.41)s) (both P<0.05). The active avoidance numbers in 10 min and 15 min exposure groups were significantly lower than that in 5 min exposure group (both P<0.05). (3) The results of immunofluorescence staining showed that the numbers of SOX2-positive cells in the four groups were statistically significant ( F=5.33, P<0.05). The SOX2-positive cells in 15 min exposure group (4.33±1.12) was significantly lower than that in control group (7.67±1.52) ( P<0.05). The numbers of NeuN-positive cells in the four groups were significantly different ( F=11.06, P<0.05), and the NeuN-positive cells in the 10 min and 15 min exposure groups((105.67±8.50), (88.33±9.50)) were significantly lower than that in the control group (127.00±6.56) ( P<0.05). The NeuN-positive cells in 15 min exposure group were significantly lower than that in 5 min exposure group (110.67±8.32) ( P<0.05). (4) Western blot results showed that the relative expression of SOX2 and NeuN proteins in the four groups was statistically significant ( F=11.560, 7.035, both P<0.05). The relative expression of SOX2 and NeuN proteins in the 15 min exposure group were significantly lower than those in control group (both P<0.05). The relative expression of SOX2 protein in 15 min exposure group was significantly lower than that in 5 min exposure group ( P<0.05). Conclusion:Acute exposure to warm pressure charge explosion simulated gas can lead to anxiety-like behavior, learning and memory deficits in rats, and significantly reduce the protein expression levels of hippocampal dentate gyrus neural stem cells and mature neuronal marker molecules SOX2 and NeuN.

Objective:To evaluated the potential developmental toxicity and teratogenicity of ammonium dinitroamide (ADN) by micromass test (MM Test) and embryonic stem cell test models.Methods:In September 2018, rat embryos were isolated and limb bud cells were collected. The limb bud cells were treated with different concentrations of ADN (0, 312.50, 625.00, 1250.00, 2500.00, 5000.00, 10000.00 μg/ml) . Half proliferation inhibitory concentration and half differentiation inhibitory concentration were calculated and the teratogenic effects were evaluated according to the criteria. For the embryonic stem cell test, the effects of different concentrations of ADN (0, 39.06, 78.13, 156.25, 312.50, 625.00, 1250.00, 2500.00 μg/ml) on the differentiation of mouse embryonic stem cells (mESCs) into myocardial cells and the cytotoxicity of mESCs and 3T3 cells were detected. The embryonic toxicity was evaluated according to the criteria. In this study, both 5-fluorouracil (5-FU) , a known strong embryonic toxic drug, and penicillin-G (P-G) , a non-embryonic toxic drug, were used to verify the effectiveness of the model, and the validated test model was applied to evaluate the embryonic toxicity of ADN.Results:In the MM Test, the inhibition rates of proliferation and differentiation of limb bud cells in ADN groups were higher than that in control group ( P<0.05) . And the half proliferation inhibitory concentration and half differentiation inhibitory concentration of ADN on limb bud cells were 7480.32 and 4526.09 μg/ml, respectively. ADN was determined to be non-teratogenic by standard. In the embryonic stem cell test, the inhibition rates of mESCs proliferation in ADN groups were higher than that in control group, and the inhibition rates of 3T3 cells in 156.25, 312.50, 625.00, 1250.00, 2500.00 μg/ml ADN groups were higher than that in control group ( P<0.05) . The half proliferation inhibitory concentration and half differentiation inhibitory concentration of ADN on mESCs were 1851.73 and 1796.39 μg/ml, respectively, and the half proliferation inhibitory concentration on 3T3 cells was 3334.35 μg/ml. ADN was determined to be non-embryotoxic by standard. Conclusion:After evaluation by MM Test and embryonic stem cell models, ADN has no embryo toxicity and is a non-teratogenic substance.

Purpose: We aimed to investigate the mechanism of phenotypic transformation of corporal cavernosum smooth muscle cells (CCSMCs) under hypoxic conditions in vitro. Materials and Methods: In this study, a hypoxia model was established using cobalt chloride (CoCl2). CCSMCs were treated with different concentrations of CoCl2 for varying time periods, and cell viability was assessed. Hypoxia-inducible factor-1α (HIF-1α), myocardin (Myocd) and phenotypic markers were detected in the CCSMCs. We also transfected the CCSMCs with si-HIF-1α and Ad-Myocd and evaluated the effects on phenotypic modulation of CCSMCs and the relationship between HIF-1α and Myocd was evaluated. Results: CoCl2 inhibited the viability of CCSMCs in a dose- and time-dependent manner, and treatment with 300 µM CoCl2 for 48 hours were the optimal conditions for establishing the hypoxia model. The results showed increased expression levels of HIF-1α and osteopontin and decreased Myocd, alpha-smooth muscle actin, and calponin levels in CCSMCs under hypoxia. HIF-1α knockdown reversed hypoxia-induced phenotypic transformation with elevated Myocd expression. Overexpression of Myocd also reversed the effect of hypoxia on the phenotypic switch, but did not affect HIF-1α expression. Conclusions Our findings showed that HIF-1α was involved in the effect of hypoxia induced by CoCl2 on CCSMC phenotypic modulation, and Myocd overexpression could inhibit this process. Thus, Myocd might be a potential therapeutic target for erectile dysfunction under hypoxia or HIF-1α activation.

Objective:To evaluated the potential developmental toxicity and teratogenicity of ammonium dinitroamide (ADN) by micromass test (MM Test) and embryonic stem cell test models.Methods:In September 2018, rat embryos were isolated and limb bud cells were collected. The limb bud cells were treated with different concentrations of ADN (0, 312.50, 625.00, 1250.00, 2500.00, 5000.00, 10000.00 μg/ml) . Half proliferation inhibitory concentration and half differentiation inhibitory concentration were calculated and the teratogenic effects were evaluated according to the criteria. For the embryonic stem cell test, the effects of different concentrations of ADN (0, 39.06, 78.13, 156.25, 312.50, 625.00, 1250.00, 2500.00 μg/ml) on the differentiation of mouse embryonic stem cells (mESCs) into myocardial cells and the cytotoxicity of mESCs and 3T3 cells were detected. The embryonic toxicity was evaluated according to the criteria. In this study, both 5-fluorouracil (5-FU) , a known strong embryonic toxic drug, and penicillin-G (P-G) , a non-embryonic toxic drug, were used to verify the effectiveness of the model, and the validated test model was applied to evaluate the embryonic toxicity of ADN.Results:In the MM Test, the inhibition rates of proliferation and differentiation of limb bud cells in ADN groups were higher than that in control group ( P<0.05) . And the half proliferation inhibitory concentration and half differentiation inhibitory concentration of ADN on limb bud cells were 7480.32 and 4526.09 μg/ml, respectively. ADN was determined to be non-teratogenic by standard. In the embryonic stem cell test, the inhibition rates of mESCs proliferation in ADN groups were higher than that in control group, and the inhibition rates of 3T3 cells in 156.25, 312.50, 625.00, 1250.00, 2500.00 μg/ml ADN groups were higher than that in control group ( P<0.05) . The half proliferation inhibitory concentration and half differentiation inhibitory concentration of ADN on mESCs were 1851.73 and 1796.39 μg/ml, respectively, and the half proliferation inhibitory concentration on 3T3 cells was 3334.35 μg/ml. ADN was determined to be non-embryotoxic by standard. Conclusion:After evaluation by MM Test and embryonic stem cell models, ADN has no embryo toxicity and is a non-teratogenic substance.

Objective To explore the value of prenatal ultrasound in diagnosis of omphalocele-exstrophy-imperforate anusspinal defects (OEIS) in first trimester.Methods Prenatal ultrasonic characteristics of 10 fetuses with OEIS complex in first trimester were retrospectively analyzed and compared with autopsy results.Results Cystic bulging in the lower anterior abdominal wall was observed in all 10 fetuses.Spinal scoliosis dysplasia was found in 10 fetuses,with myelomeningocele in 3 fetuses.No normal bladder was visualized in 8 fetuses.Thickened nuchal translucency was noticed in 5 fetuses,among which neck lymphatic hydrocele was found in 1 fetus.The bilateral clubbed feet and left lower mutilation was observed in 1 fetus,respectively.All 10 OEIS complex fetuses were found accompanied with short umbilical cord,while single umbilical artery and umbilical cord cyst were found in 4 and 1 fetus,respectively.Autopsy showed abdominal wall defects with exstrophy in 10 fetuses.However,no complete cystic bulging was found.Besides,autopsy also showed pubic symphysis separation and bladder exstrophy in 10 fetuses without obvious genitalia nor anus.Conclusion Cystic bulging in the lower anterior abdominal wall is the most common prenatal ultrasonic characteristic of OEIS complex in first trimester.

Objective:To investigate clinical efficacy of parasacral perforator flap (PPF) on postoperative wound healing in pilonidal sinus diseases (PSDs).Methods:The surgery steps were as follows: (1) To preoperatively detect parasacral perforator arteries with the handhold Doppler probe and mark them; (2) To remove the infected and necrotic tissues of PSDs completely; (3) To design the PPF according to the wound size and the parasacral perforator arteries' localization; (4) To harvest the flap from the gluteus maximus muscle surface and transfer it to the wound without tension. Several data were documented, including surgical duration, flap length, flap width, drainage tube placement duration, hospital stay, duration from operation to stitch removal, postsurgical complications and recurrence.Results:There were six patients with PSDs whose postoperative wound healing was repaired by PPF, admitted in our department from March 2021 to March 2023. Of them, five were male and one was female. Their median age was 24 (range: 18-33) years old. Their median surgical duration was 165 (range: 134-207) minutes, median length of PPF was 8 (range: 7-11) cm, median width of PPF was 3 (range: 3-4) cm, mean duration of drainage tube placement was 8 (range: 4-17) days, mean hospital stay was 13 (range: 6-23) days, mean duration from operation to stitch removal was 14 (range: 14-17) days, median follow-up time was 6-16 months. Incisions of all six cases achieved first-intention healing without early- or late-stage complications. No recurrence occurred during follow-up. All patients involved were satisfied with their clinical efficacy.Conclusion:The utility of PPF in postoperative wound healing of PPDs was effective, safe and reliable.

Objective:To explore the effect of nerve injury in rats by neurobehavioral experiments, in order to provide a model and idea for further clarification of the traumatic brain injury mechanism under explosion exposure.Methods:From May 2021 to August 2022, 160 SPF male rats were randomly divided into four groups, including control group, 60 kPa group (low intensity group), 90 kPa group (medium intensity group) and 120 kPa group (high intensity group). The blast induced traumatic brain injury (bTBI) model of rats was established by using the shock tube platform to simulate the shock wave parameters of the explosion overpressure of 60 kPa, 90 kPa and 120 kPa. Acute observation was carried out after 24 h and 7 d of explosive exposure, and chronic recovery observation was carried out after 28 d and 90 d. The time effect of shock wave brain injury in different situations was discussed by open field, light dark test, active avoidance test. Finally, the results of brain injury in rats were detected by pathological tissue staining.Results:After 24 h explosion exposure, compared with the control group, the rest time of rats in low and high intensity groups increased, the total movement distance decreased, and the number of visits to the camera obscura decreased, with statistical significance ( P<0.05). After 7 days of exposure, compared with the control group, the rest time of rats in high intensity group increased, and the number of visits to the obscura decreased, with statistical significance ( P<0.05). After 28 and 90 days of exposure, compared with the control group, there were no significant differences in rest time, total exercise distance and times of visiting the camera obscura in all intensity groups ( P>0.05). After 24 h of explosive exposure, compared with the control group, the cell morphology of rats in each intensity group was normal, and no inflammatory cell infiltration was observed. Conclusion:In the acute phase (24 h) of blast exposure, rats have no desire to explore the outside world, and shock wave exposure may damage the neurological function of rats.