[Study on reproductive toxicity of nano-cadmium sulfide with different particle sizes on male mice].

Objective: To investigate the reproductive toxicity of cadmium sulfide nanoparticles (Nano-CdS) with different particle sizes on male mice. Methods: In January 2019, 30 SPF grade male mice were randomly divided into a control group, an experimental group[CdS Ⅰ group (particle size approximately 5 nm), and a CdS Ⅱ group (particle size approximately 50 nm) ], with 10 mice in each group. The experimental group was orally gavaged with 100 mg/kg, once a day, while the control group was gavaged with an equal volume of physiological saline for 45 consecutive days. After 45 days, levels of cadmium accumulation in testis were determined directly by AAS, deformity and testicular histopathological changes were also observed. Serum testosterone levels were measured by enzyme-linked immunosorbentassay (ELISA), expression levels of P450scc, 17ß-HSD and P450c17 mRNA were determined by real-time PCR. P450c17 protein was determinated by Western Blot. Results: The histopathological results showed that the testes of the experimental group mice showed varying degrees of damage; Ultrastructural observation showed that the ultrastructure of mouse testicular cells in each experimental group showed varying degrees of mitochondrial expansion and disappearance of cristae, as well as irregular nuclear membranes. The degree of damage in CdS Ⅰ group was milder than that in CdS Ⅱ group. Compared with the control group, the cadmium content in the testes of the CdS Ⅰ and CdS Ⅱ groups significantly increased (P=0.001, 0.001), and the CdS Ⅱ group was higher than the CdS Ⅰ group (P=0.001). Compared with the control group, the levels of testosterone in the CdS Ⅰ and CdS Ⅱ groups decreased with statistical significance (P=0.001, 0.001). Real time fluorescence quantitative PCR results showed that compared with the control group, the experimental group's P450scc, 17ß-HSD. The expression levels of 17ß-HSD and P450c17 mRNA were significantly reduced, with statistically significant differences (P=0.001, 0.001, 0.001), and CdS Ⅱ group 17ß-HSD. The expression levels of 17ß-HSD and P450c17 mRNA were significantly lower than those of CdS Ⅰ group (P=0.001, 0.036). The Western Blot assay results showed that the expression levels of P450c17 protein in the testes of CdS Ⅰ and CdS Ⅱ groups of mice were significantly reduced, with statistical significance (P=0.001, 0.001) ; And the CdS Ⅱ group was significantly lower than the CdS Ⅰ group (P=0.001). According to Spearman correlation analysis, testosterone levels are correlated with P450scc, P450c17, 17ß-HSD mRNA. There is a highly positive correlation between 17ß-HSD mRNA levels, with statistically significant differences (r(s)=0.88, 0.80, 0.70, P=0.001, 0.001, 0.004) . Conclusion: Nano cadmium sulfide may induce reproductive toxicity by reducing the expression levels of key enzyme genes and enzyme protein activity in testosterone and its synthesis in mice, and the CdS Ⅱ group has a stronger toxic effect.

[Genetic analysis of varicella-zoster virus in patients with viral anterior uveitis].

Objective: To analyze the intraocular varicella-zoster virus (VZV) infection and genetic characteristics in patients clinically diagnosed with viral anterior uveitis. Methods: A total of 83 aqueous humor samples were collected from patients clinically diagnosed with viral anterior uveitis infection in China from June 2018 to July 2019. The positive samples infected with VZV were screened by real time polymerase chain reaction, and the single nucleotide polymorphisms on the open reading frames 22, 38 and 62 of the positive samples were amplified and analyzed. According to the gene characteristics of the amplified target fragment, the vaccine strain and wild strain (8 vaccine strains and the rest were wild strains) were identified to determine the genotype. Results: There were 83 patients (31 females and 52 males) with viral uveitis infection, whose mean age was 51.0 (45.5, 61.0) (range: 15-83) years,, and, of which 57.8% (48 cases) were infected with viral uveitis over 50 years of age. None of the patients had a history of varicella or herpes zoster vaccination. Of the samples of 83 patients infected with viral uveitis, 57 (68.6%) were positive for VZV. Among them, 14 were successfully amplified to obtain the target fragment gene sequences, all of which were wild strains by analysis, and belonged to Clade2 of genotype, which was the same as the VZV vaccine strain types infected by varicella and herpes zoster patients in China. Conclusion: From 2018 to 2019, VZV infection in Chinese patients with viral anterior uveitis was a wild strain, and the genotype belonged to Clade2 as the vaccine strain, which was the same as the main epidemic genotype of VZV infection in Chinese patients with varicella and herpes zoster.

Biological background of the genomic variations of cf-DNA in healthy individuals.

BACKGROUND: Cell-free DNA (cf-DNA)-based liquid biopsy is emerging as a revolutionary new method in individualized cancer treatment and prognosis monitoring, although detecting early-stage cancers using cf-DNA remains challenging, partially because of the undefined biological background of cf-DNA. MATERIALS AND METHODS: We investigated somatic mutations in the cf-DNA of 259 cancer-free individuals with a median age of 47 years using an endogenous barcoding duplex method with an ultralow base error rate (2 × 10-7) and compared the variant allele frequencies (VAFs) of these mutations between the cf-DNA and the corresponding blood cell DNA. RESULTS: Sixty percent (155/259) of the samples showed at least one nonsynonymous mutation on either of two similar target panels covering 508 and 559 cancer-related genes. For individuals older than 50 years of age, the positive rate increased to 76%. Most cf-DNA mutations were also present at similar VAFs in the paired blood cell DNA. The most frequently mutated genes were driver genes of hematologic malignancies, including DNMT3A, TET2, AXSL1, and JAK2. However, the other 58.4% (192/329) of the mutations were likely 'passenger mutations' of clonal hematopoiesis, including mutations in NOTCH2, FAT3, EXT2, ERBB4, and ARID2, which are driver genes of solid tumors. CONCLUSION: Hematopoietic clone-derived mutations, including 'driver mutations' and 'passenger mutations', are prevalent in the cf-DNA of both healthy individuals and cancer patients and may be a potential source of false positives in the liquid biopsy. Our results also suggest the ineffectiveness for distinguishing clonal hematopoietic mutations of low VAF (≤0.1%) from tumor-derived mutations using conventional next-generation sequencing of blood cell DNA. However, an error correction model with an ultralow error rate and high coverage depth is required for blood cell DNA sequencing, which is difficult and costly to achieve with current technologies.

[Surgical treatment of port-site metastases after laparoscopic radical resection of gastric cancer].

Objective: To investigate the feasibility of surgical treatment of port-site metastasis after laparoscopic radical resection of gastric cancer. Methods: The clinical and follow-up data of five patients with port-site metastases after laparoscopic radical resection of gastric cancer at Zhejiang Provincial People's Hospital between January 2014 and January 2018 were retrospectively analyzed. Results: Port-site metastases occurred within 6 months after gastrointestinal tumor resection in three patients, 10 months after the operation in one patient, and 30 months after the operation in one patient, respectively. Metastasis to the abdominal cavity or distant metastasis was excluded before the surgical treatment of the port-site metastases, and all patients recovered well after the operation. No incisional infection or hernia occurred. By December 2018, two patients died (they survived for 13 and 24 months, respectively) and three patients survived. The follow-up duration ranged from 7 to 19 months. Conclusions: Surgical resection of port-site metastases is not difficult due to their superficial location. Surgical treatment can improve the prognosis of patients without abdominal or distant metastasis/recurrence.

[Preliminary application of endoscopic titanium clip localization combined with three-dimensional CT reconstruction in the determination of resection margin of gastric central cancer under laparoscopy].

Objective: To evaluate the accuracy of endoscopic titanium clip localization combined with CT three-dimensional reconstruction for the control of incision margin in early gastric cancer under laparoscopy. Methods: A prospective analysis was made for gastric cancer whose lesions were located in the middle of the stomach and T stage was 1 to 2 from October 2017 to January 2019 at Department of Gastrointestinal and Pancreatic Surgery, Zhejiang Provincial People's Hospital. Totally 25 patients were eventually enrolled in the study. There were 17 males and 8 females aging of (63.6± 7.2) years (range: 48 to 77 years). All cases were treated with titanium clip localization under endoscope combined with CT three-dimensional (3D) reconstruction to construct a virtual panorama of gastric cavity and lesions, and to design surgical margins. Laparoscopic surgical resection was performed according to the surgical margins designed before operation. The distance from the gastric angle to the origin of the minor curvature of the incisional margin, the distance from the gastric angle to the the center of lesion and the distance of the upper incision margin were measured under three-dimensional CT reconstruction and under actual specimen. Paired t test was used to compare the three distances measured by two methods. Results: The measured distances from the gastric angle to the center of the lesion and the proximal incisional margin under 3D reconstruction CT were according to the measured values of actual specimens ((2.67±1.38) cm vs. (2.83±1.56) cm, t=1.51, P=0.14; (5.23±0.60) cm vs. 5 cm, t=1.93, P=0.07); the measured distances from the gastric angle to the origin of the minor curvature of the incisional margin under CT 3D reconstruction were different with the measured values of solid specimens ((5.94±0.94) cm vs. (6.37±0.90) cm, t=3.52, P=0.00). Conclusion: The method of titanium clip localization combined with CT 3D reconstruction can provide a feasible laparoscopic localization method and incision edge solution for T1 to 2 gastric central cancer.

[Laparoscopic radical antegrade modular pancreatosplenectomy for left-sided pancreatic adenocarcinoma: analysis of 12 cases].

Objective: To evaluate the safety and feasibility of laparoscopic radical antegrade modular pancreatosplenectomy(Lap-RAMPS) for left-sided pancreatic adenocarcinoma. Methods: Clinical data of total 12 patients underwent Lap-RAMPS for left-sided pancreatic adenocarcinoma at Department of Gastrointestinal and Pancreatic Surgery, Zhejiang Provincial People's Hospital from March 2016 to August 2017 were reviewed retrospectively.There were 7 male patients and 5 female patients, with median age of 60.5 years old(47-68 years old). Abdominal enhanced CT, pancreatic MRI, PET-CT were performed on all patients to evaluate the lesion and exclude metastasis.Follow-up were done with out-patient clinic or telephone consultancy until October 2017. Results: All patients underwent pure Lap-RAMPS.The medium operative time was 250 minutes(180-445 minutes), and the blood loss was 150 ml(50-500 ml). The medium first flatus time and diet resumption time were 3.0 days(1-5 days) and 3.5 days(1-7 days) respectively.The medium postoperative hospital stay was 9 days(4-18 days). Morbidity occurred in 8 patients with gastric empty delay(n=1), bleeding(n=1), fluid collection(n=3). There was no mortality.The medium overall number of retrived lymph nodes was 15.6 and the positive rate was 41.7%. The R0 rate was 100%.The medium follow-up was 10 months.One patient was diagnosed as liver metastasis after 8 months and accepted chemotherapy.One patient died after 14 months for tumor recurrence and metastasis.Others survived without tumor recurrence or metasitasis. Conclusion: Lap-RAMPS is safe and feasible with accepted oncological outcomes for selected left side pancreatic adenocarcinoma under skilled hands.

[Strategy in management of adjacent organ lesion during laparoscopic pancreaticoduodenectomy].

Objective: To evaluate the safety and feasible of adjacent organ resection during laparoscopic pancreaticoduodenectomy(LPD), and summary the surgical strategies. Methods: Clinical data of 15 adjacent organ resections combined with LPD from March 2013 to September 2017 were reviewed.There were 10 male and 5 female patients aging from 20 to 86 years, and the body mass index ranged from 19.6 to 34.5 kg/m(2).Two patients had previous abdominal surgical history.Two patients underwent preoperative chemotherapy. Results: The resected adjacent organs included liver(n=4), stomach(n=3), colon(n=6), right kidney with embolectomy and vasoplastic of inferior vena cava(n=1), and spleen artery aneurysms(n=1). The operative time ranged from 280 to 450 minutes, and the blood loss ranged from 100 to 450 ml.The total complication rate was 5/15 and no one died in 90 days after surgery.The postoperative hospital stay ranged from 10 to 42 days with medium 18 days.The pathology included adenocarcinoma of stomach and duodenum(n=1), gastric cancer invading pancreas or duodenum(n=2), ampullary adenocarcinoma with left hepatolithiasis(n=1), ampullary adenocarcinoma with a benign lesion in left liver(n=1), ampullary adenocarcinoma with single liver metastasis(n=1), ampullary adenocarcinoma(n=1), pancreatic intraductal papillary mucinous neoplasm with splenic artery aneurysms(n=1), pancreatic neuroendocrine neoplasm with colon cancer(n=1), distal common bile duct adenocarcinoma involving righ hepatic duct(n=1), pancreatic neuroendocrine neoplasm invading inferior vena cava and right renal vein(n=1), duodenal adnocarcinoma(n=1), duodenal ewing's sarcoma(n=1), duodenal intesititialoma(n=2). The follow-up was from 3 to 40 months with the medium survival of 17.5 months. Conclusions: The oncological outcomes of PD combined with adjacent organ resection is acceptable.Surgical treatment for those patients with periampullary neoplasma and adjacent organ lesions should be aggressive.

[Reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol in mice: an experimental study].

Objective: To investigate the reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol in mice and to provide a basis for its risk assessment. Methods: The reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol was tested using the screening method of chemicals with reproductive and developmental toxicity in "Chemical Testing Method" (SEPA). After five days of adaptive feeding, 120 specific pathogen-free healthy Kunming mice (male/female ratio=1:1) were orally administered 0 (control) , 146, 292, and 584 mg/kg 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol for two weeks. One male mouse was mated with one female mouse in a single cage. The day on which a vaginal plug was observed was defined as gestation day 0 (GD0). The exposure for female mice was sustained to four days postpartum and the exposure for male mice was sustained for two weeks after mating. The body weight, food intake, body length, tail length, and sex ratio were recorded and the reproductive index was calculated. The reproductive organs were weighed and subjected to histopathological examination. Results: The 584 mg/kg group had significantly lower body weight at weeks 5 and 6 and food intake at week 6 in male mice, uterus weight and uterus/body weight ratio in female mice, and body weight, body length, and tail length on day 0 in offspring compared with the control group (all P<0.05). The 292 mg/kg group had significantly lower testis weight of male mice and food intake of female mice at gestational week 2 than the control group (both P<0.05). The 146 mg/kg group had significantly lower food intake of female mice at gestational week 2 than the control group (P<0.05) . Conclusion: For male and female Kunming mice, the no observed adverse effect levels of 2- (2H-1, 2, 3, -benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol are both 146 mg/kg.

[Autophagy regulated by JWA influenced sensitivity of esophageal cancer to cisplatin].

Objective: To explore the effect of JWA on cisplatin sensitivity and its potential molecular mechanism in esophageal cancer. Methods: The siRNA was used to inhibit the JWA expression, then cisplatin sensitivity and LC3 (autophagy related protein) expression levels were observed in TE1 cells.Further, the effect of autophagy inhibitor tamoxifen (3-MA) on above process was determined.Cisplatin sensitivity of 20 fresh esophageal cancer samples was evaluated by histoculture drug response assay (HDRA). Result: Silencing JWA gene increased the sensitivity of TE1 cells to cisplatin (P<0.05), and decreased the LC3-Ⅰ and LC3-Ⅱ proteins induced by cisplatin.Furthermore, combined with 3-MA increased the inhibition rate of cisplatin in JWA silencing group (P<0.05). Additionally, the inhibition rate of cisplatin on tissues with low JWA expression were higher than those with high expression (45.6% vs 25.6%, P=0.005). Conclusions: JWA could influence the cisplatin sensitivity by regulating autophagy in esophageal cancer.

[Expression characteristics of PTEN and NDRG1 in colorectal carcinoma and their prognostic value].

Objective: To study the expression status and clinical significance of PTEN and NDRG1 in colorectal carcinoma. Methods: Tissue samples of 91 colorectal cancers, 30 colorectal adenomas and 21 colorectal normal mucosa tissues were collected. Postoperative specimens were examined by immunohistochemistry for PTEN and NDRG1 expression. The expression of PTEN and NDRG1 was correlated with clinicopathological feature. Results: The expression of PTEN and NDRG1 in the studied cases was detected in 55.0%(50/91) and 76.9%(70/91), respectively. Their expression was significantly different from that of colorectal adenomas and normal colorectal mucosa tissues(P<0.05). Decreased expression of PTEN and over expression of NDRG1 were significantly related to the lymph node metastasis (P<0.05). The expression of PTEN was negatively related to that of NDRG1 in colorectal carcinoma(rs'=-0.251, P=0.016). The patients with negative expression of PTEN showed a lower disease free survival and overall survival(P<0.05). Conclusions: Loss of expression of PTEN protein may be an important molecular marker in predicting the occurrence and PTEN may be useful as a prognostic marker of colorectal carcinoma. NDRG1 plays a role in the development of colorectal carcinoma, although not a prognostic indicator.The ancillary study with combined detection of PTEN and NDRG1 may be useful in difficult cases.

Proposal of a prediction score for hematoma expansion after intracerebral hemorrhage.

OBJECTIVE: To propose and validate a prediction score for intracerebral hemorrhage (ICH) patients at risk of hematoma expansion (HE). DESIGN: A retrospective observational study was designed to propose and validate the score. SETTING: Sanxiang Road branch and Xuguan branch belonging to the Second Affiliated Hospital of Soochow University (China). PATIENTS: A total of 317 ICH patients in Sanxiang Road branch were registered as the development cohort, and 109 ICH patients in Xuguan branch were enrolled as the validation cohort. PROCEDURE: Independent risk factors for HE were identified using multiple logistic regression analysis. A prediction score was then proposed based on ß coefficients and preliminarily verified in the validation cohort. MAIN VARIABLES: All clinical data of the patients were compiled from the electronic medical records. Hematoma expansion was defined as an increase in hematoma volume >33% or absolute hematoma growth >6ml from the initial scan. Specific non-contrast CT(NCCT) signs were identified by two observers independently. RESULTS: Our score demonstrated satisfactory discrimination ability for HE (area under the ROC curve 0.854 in the development cohort versus 0.893 in the validation cohort). Appropriate calibration was found in the development cohort, whereas calibration in the validation cohort was slightly lower but still within the accuracy range (maximum deviation, average deviation and P were 0.070, 0.028, 0.773, respectively, versus 0.114, 0.056, 0.156). Decision curve analysis of the score from two samples were both far from the curve of treat all and curve of treat none, which verified its security and reliability. Patients with a total score ≥4.5 were at greatest risk of HE. CONCLUSION: The score may provide some reference and help in accurately identifying individuals at high risk of HE, allowing rapid guidance of clinical management and also serving as an aid in clinical trials.

Long-lasting infiltration anaesthesia by lidocaine-loaded biodegradable nanoparticles in hydrogel in rats.

BACKGROUND: Infiltration of a long-lasting anaesthetic is helpful during the post-operative period. The recently developed local drug delivery system, biodegradable nanoparticles in a thermo-sensitive hydrogel (nanogel system), may possibly provide an extended duration of drugs. Therefore, we evaluated whether prolonged infiltration anaesthesia could be achieved by loading lidocaine into this delivery system. METHODS: Thirty male rats were randomized into five groups of six rats each: saline; 2% hydrochloride lidocaine solution; lidocaine-loaded nanogel system and its compositing formulations, namely lido-nano gel; lido-nano; and lidogel. Durations of local anaesthesia with subcutaneously injected agents were measured by tail flick latency tests in a randomized, blind fashion. RESULTS: Lido-nano gel produced effective anaesthesia for 360+/-113 min, compared with 150+/-33 min by lidogel, 180+/-37 min by lido-nano, and 110+/-45 min by lidocaine solution (P<0.001, means+/-SD), and elicited complete sensory blockade for 300+/-114 min, compared with 75+/-37 min by lidogel, 105+/-53 min by lido-nano, and 60+/-33 min by lidocaine solution (P<0.001, means+/-SD) without severe skin/systemic toxicity. CONCLUSION: Lidocaine-loaded biodegradable nanoparticles in hydrogel produced prolonged infiltration anaesthesia in rats without severe toxicity, indicating a possible way to develop long-lasting local anaesthetics.

Potential roles of lncRNA-Cox2 and EGR1 in regulating epidural fibrosis following laminectomy.

OBJECTIVE: Epidural fibrosis, one of the common complications after spinal surgery, seriously affects the surgical decompression effect. Effectively inhibiting the fibrous tissue hyperplasia is pivotal to reduce the scar adhesion. Previous studies showed that early growth response 1 (EGR1) is associated with the fibroblast reactivity induced by transforming growth factor-beta (TGF-ß) and plays a vital regulatory role in scar formation; however, the upstream targets and mechanisms still remain unclear. In this work, it was found that the level of long non-coding ribonucleic acid (lncRNA)-cyclooxygenase-2 (COX2) was significantly negatively correlated with EGR1 expression and the severity of the scar. Therefore, it was conjectured that lncRNA-COX2 may decrease fibroplasia and scar formation by negatively regulating EGR1. MATERIALS AND METHODS: TGF-ß was used to activate the embryonic and adult rat fibroblasts. Rats underwent laminectomy to establish the epidural fibrosis model. The changes in the levels of fibroplasia-related genes were measured and analyzed through messenger RNA (mRNA), lncRNA, and micro RNA expression profile chips. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was applied to determine the levels of EGR1 and lncRNA-COX2, and Western blotting was adopted to detect the content of EGR1, collagen I (Col-1), Col-3, and alpha-smooth muscle actin (α-SMA). The scar formation was reflected by hematoxylin and eosin (HE) staining and Masson staining, and the expression level of α-SMA in the scar tissues was measured via immunohistochemistry. Finally, micro-magnetic resonance imaging (MRI) was utilized to examine the different degrees of epidural fibroplasia. RESULTS: It was found that the reactivity of embryonic rat fibroblasts to the TGF-ß stimulation was different from that of adult rat fibroblasts. LncRNA-COX2 was highly expressed in the embryonic rat fibroblasts, but lowly expressed in the adult rat fibroblasts, which had negative correlations with the EGR1 level in embryonic and adult rat fibroblasts. In addition, it was revealed that the expression of EGR1 in the adult rat fibroblasts was remarkably higher than that in the embryonic rat fibroblasts after the activation with TGF-ß. Meanwhile, the level of lncRNA-COX2 was lowered after the activation, especially in the adult rat fibroblasts. It was discovered in the in-vivo model that the degree of fibroplasia was positively associated with EGR1 level and negatively correlated with lncRNA-COX2 level. CONCLUSIONS: The results of this research elucidated that the down-regulation of lncRNA-COX2 is involved in the epidural scar formation and related to the elevated EGR1 level which regulates the activation of fibroblasts and secretion of massive extracellular matrixes, suggesting that lncRNA-COX2 may modulate the role of fibroblasts in scar formation as an upstream action target of EGR1.

Biomechanical strength impact of lateral wall breach on spinal pedicle screw fixation.

OBJECTIVE: The purpose of our study was to make a comparison between the fixation strength of optimum placed pedicle screw (OS) and re-directionally accurate placed pedicle screw (RS) after lateral pedicle breach. PATIENTS AND METHODS: A total of 30 fresh lumbar vertebrae (L1-5) were gained from 6 male or female pigs weighing about 100 kg, which were divided into 2 groups according to different ways of pedicle screws placement: OS group (n=30) and RS group (n=30). MTS machine was employed to detect the screw loosening and axial pullout. We examined seating torque, screw-loosening force, the maximal torque and post-loosening axial pullout in each pedicle screw. RESULTS: Maximal insertion torque of OS was (111.6±8.4) N•cm and RS was (79.0±6.3) N•cm, which indicated a significant difference (Z=3.012, p=0.003). Seating torque of OS and RS were (85.9±5.6) N•cm and (60.3±4.8) N•cm separately, and the difference was statistically significant (Z=2.799, p=0.006). Screw loosening force of OS and RS were (75.9±7.0) N and (52.4±6.3) N respectively, and the difference was statistically significant (Z=2.652, p=0.003). Post-loosening axial pullout force of OS and RS were (328.5±11.3) N and (269.1±9.6) N separately, demonstrating that the difference was statistically significant (Z=2.865, p=0.004). CONCLUSIONS: RS placement is an alternative for remediation following a lateral wall breach evidenced by significantly decreased seating torque, screw loosening force, the maximal torque and post-loosening axial pullout compared with OS.

Crocetin improves endothelium-dependent relaxation of thoracic aorta in hypercholesterolemic rabbit by increasing eNOS activity.

Our previous studies have proven that crocetin (CCT), extracted from Gardenia jasminoides Ellis, possesses the anti-atherosclerotic effect. Because endothelial dysfunction strongly contributes to the initiation and progression of atherosclerosis, the present study aims to investigate whether CCT is capable of improving this dysfunction and to explore the possible mechanisms. Endothelial dysfunction was induced by in vivo feeding high cholesterol diet (HCD) to rabbit and by in vitro treating bovine aortic endothelial cells (BAECs) with oxidized LDL (oxLDL). Endothelium-dependent relaxation (EDR) evoked by acetylcholine (Ach) and endothelium-independent relaxation (RIDR) mediated by sodium nitroprusside (SNP) of thoracic aorta isolated from rabbit were measured. The results indicated that the EDR in HCD alone treated rabbits was seriously impaired and the maximal relaxation induced by Ach (10(-5.5) M) was only 54% that in control rabbit fed with regular diet. Oral complementation with CCT (15, 30 mg/kg) dose-dependently improved this impairment and restored the maximal relaxation to 68% and 80% that in control group, respectively. However, the EIDR maintained comparable in all groups. Complementation with CCT (15, 30 mg/kg) simultaneously increased serum level of nitric oxide (NO), upregulated vessel activity and mRNA expression of endothelial NO synthase (eNOS) as well as vessel cyclic GMP (cGMP) content compared with those in rabbit treated with HCD alone. Inducible NOS (iNOS) activity remained unchangeable in all groups. In BAECs, oxLDL treatment decreased NO production, downregulated both activity and mRNA expression of eNOS. While those decrease or downregulation were inhibited by co-treatment with CCT (0.1, 1, 10 microM) in a dose-dependent manner. These findings suggested that CCT significantly restored the EDR of thoracic aorta in hypercholesterolemic rabbit, which might be explained by its action to increase the vessel eNOS activity, leading to elevation of NO production.

Isolation and characterization of sheep lactoferrin, an inhibitor of platelet aggregation and comparison with human lactoferrin.

Highly purified sheep lactoferrin was isolated from ovine whey in a single chromatographic step (FPLC): it was characterized by electrophoresis, N-terminal sequence determination and compared with lactoferrins from other species. Sheep and human lactoferrins inhibited thrombin-induced platelet aggregation (median inhibitory concentration: IC50 5 and 4 microM, respectively). Pepsin hydrolysates of human and sheep lactoferrins were fractionated by reverse-phase high-performance liquid chromatography and only one peak was an inhibitor of platelet aggregation. The sheep or human lactoferrin binding to platelets was studied.

Sheep kappa-casein peptides inhibit platelet aggregation.

The C-terminal part (residues 106-171) of sheep kappa-casein, called caseinoglycopeptide (CGP), inhibits thrombin- and collagen-induced platelet aggregation in a dose-dependent manner (mean inhibitory concentration (IC50) 215 microM and 100 microM, respectively). An enzymatic hydrolysate of CGP was fractionated by reverse phase high performance liquid chromatography: three peptides KDQDK (residues 112-116), TAQVTSTEV (residues 163-171) and QVTSTEV (residues 165-171) completely inhibited thrombin-induced platelet aggregation. CGP at a concentration near its IC50 had a very long life when incubated in human or guinea-pig plasma. An ex vivo experiment showed that 17% of CGP was found 60 min after its i.v. bolus injection in guinea-pig. By hydrophobic cluster analysis, human fibrinogen and sheep kappa-casein peptides, inhibitors of platelet aggregation, were compared and we observed similarities for their C-terminal parts and for their short peptides (RGDF and KDQDK).

Polyamine profiles in rat testis, germ cells and Sertoli cells during testicular maturation.

Polyamine cellular concentrations (putrescine, spermidine and spermine) in the rat testis and testicular cell types were determined by fluorescence spectroscopy of their dansyl derivatives. A method is described to separate dansylated polyamines by high performance liquid chromatography in less than 12 minutes. In rat Sertoli cells, polyamine concentrations (per mg DNA) were greater than those in germ cells and the testis. The concentrations of all three polyamines increased with age. Concentrations of spermidine and spermine in germ cells also increased with age and leveled off after 27 to 35 days. On the other hand, higher putrescine levels were found in the testis of young rats (13 to 22 days) while the greatest spermidine and spermine contents were observed in the testis from rats of 31 to 35 days old. Of great interest, Sertoli cells from all age groups studied released a relatively large quantity of putrescine and a smaller amount of spermidine, but no spermine, into culture media. The amount of polyamine released by Sertoli cells varied with the age of the animal. Sertoli cells from 27-day-old rats released the greatest quantity of putrescine on a per mg DNA basis. The release of putrescine increased after hypotonic treatment that removed contaminating germ cells from the remaining Sertoli cells. It is concluded that cellular polyamine levels in the rat testis, germ cells and cultured Sertoli cells and the amount of polyamines released by Sertoli cells were age-dependent during the first wave of spermatogenesis.

Protective effects of ulinastatin on intestinal injury during the perioperative period of acute superior mesenteric artery ischemia.

OBJECTIVE: This work aims to explore the protective effects of ulinastatin on intestinal injury during the perioperative period of acute superior mesenteric artery ischemia (ASMAI). PATIENTS AND METHODS: 28 patients undergoing revascularization were divided into 2 groups, with 14 cases each. The cases in the observation group (OG) were treated with ulinastatin 300,000 U intravenously 30 min before the operation, and continuously treated with 300,000 U every 4 hr thereafter until 24 hr of the operation, while those in the control group (CG) were not given the intervention of ulinastatin. Patients' circular intestinal fatty acid binding protein (I-FABP) levels were measured at the following time points to reflect the intestinal injury: 30 min before the operation, before revascularization, then 1, 12 and 24 hr after the operation. The white blood cell counting (WBC), serum alanine aminotransferase (ALT), serum creatinine (Cr), D-dimer, and serum endotoxin (ET) were also measured simultaneously for the analysis of the significance of their values with the intestinal injury. RESULTS: There were no significant differences (p > 0.05) in ischemia duration, length of the affected intestinal segments, WBC, ALT and Cr levels at the above time points between the 2 groups, and all the indicators of the 2 groups, including the mean circular I-FABP levels before the operation and the revascularization, showed no significant difference (p > 0.05). After the blood supply was restored, the I-FABP levels in OG dropped significantly as compared with those in CG. The pattern of circular ET levels appeared the similar manner as the circular I-FABP levels did. CONCLUSIONS: Our study showed a protective effects of ulinastatin on intestinal injury during the perioperative period of ASMAI, as revealed by the circular I-FABP levels which mainly happened after the blood supply was restored.

Down-regulation of cyclooxygenase-2 is involved in ischemic postconditioning protection against renal ischemia reperfusion injury in rats.

Ischemic postconditioning (IPostC) is a phenomenon whereby rapid intermittent interruptions of blood flow in the early phase of reperfusion protect an organ from ischemia-reperfusion injury. In the present study, we investigated whether the protective effect of IPostC was associated with the cyclooxygenase-2 (COX-2) pathway by evaluating its expression following renal ischemia-reperfusion in rats. Animals underwent 45 minutes of renal pedicle occlusion followed by reperfusion for 1.5, 3, 6, 12, or 24 hours. IPostC was performed by six 10-second cycles of reperfusion and 10 seconds of renal pedicle occlusion at the end of ischemia. Blood and kidney samples were collected at each reperfusion time point. The protein expression of COX-1 and COX-2 were evaluated by Western blotting. Our data showed that IPostC attenuated the renal dysfunction and decreased COX-2 expression induced by ischemia-reperfusion insults. The results indicated that the protective effect of IPostC was related to down-regulation of COX-2 expression.