AIM: To investigate the efficacy and safety of ZKY001 eye drops with different concentrations in the treatment of corneal epithelial defects(CED)after primary pterygium excision.METHODS: This was a multicenter, randomized, double-blinded, placebo-controlled phase II clinical trial. From March 15, 2022 to November 14, 2022, patients with primary pterygium who had undergone surgery were recruited from 12 tertiary hospitals across China. Using block randomization, 178 patients(178 eyes)were randomly assigned to 3 groups in a 1:1:1 ratio: 0.002% ZKY001 group(n=59), 0.004% ZKY001 group(n=59), and placebo group(n=60, receiving ZKY001 sham eye drops). Subjects in each group received 1 drop of the study drug 4 times per day for 4 d. The percentage of CED area recovery from baseline, the first complete healing time of CED area, the number of first complete healing cases of CED, and changes in visual analogue scale(VAS)scores for eye discomfort including eye pain, foreign body sensation, tearing and photophobia were observed.RESULTS: In terms of improvement in CED, there were no statistically significant differences among the three groups including the first healing time of CED, the percentage improvement in CED area compared to baseline, and the percentage of first healing cases at different follow-up visits(all P>0.05). Numerically, the first healing time of CED was shorter in the test groups compared to the placebo group(67.87±21.688 h for the 0.002% ZKY001 group, 61.48±22.091 h for the 0.004% ZKY001 group, and 68.85±20.851 h for the placebo group). On D1 morning, the percentage improvement in CED area compared to baseline was maximally different from the placebo group, and the numerical difference advantage was maintained at subsequent follow-up visits. The number of first healing cases in the CED area at different follow-up visits was higher in the test groups than the placebo group. In terms of improvement in ocular discomfort, the total VAS scores were lower in the test groups compared to the placebo group, mainly due to reductions in foreign body sensation and pain scores. At D3, the 0.004% ZKY001 group showed statistically significant improvement in foreign body sensation(P<0.017). In terms of safety, the overall incidence of adverse events was low(9.0%)and similar among groups.CONCLUSION: The use of ZKY001 eyedrops after primary pterygium surgery can safely improve the CED repair, and alleviate postoperative symptoms caused by CED.

ObjectiveTo apply ultrasonography to quantitatively evaluate the structure and biomechanical properties of gastrocnemius muscle in children with spastic cerebral palsy. MethodsFrom December, 2020 to December, 2021, 36 children with spastic diplegia or hemiplegia were selected as experimental group, and other 30 healthy children matched age and gender were as control group during the same period. All the subjects were measured muscle thickness (MT), muscle fiber length (FL), pinnate angle (PA) and shear wave velocity (SWV) with two-dimensional ultrasound imaging and shear wave elastography on the medial gastrocnemius of the affected side (hemiplegia), more serious side (diplegia) or random side (control group), while the gastrocnemius muscle tone and gross motor function of the experimental group were assessed with modified Ashworth Scale (MAS) and Gross Motor Function Measure (GMFM). ResultsMT and FL were less in the experimental group than in the control group (|t| > 3.937, P < 0.001), while PA and SWV were more (|t| > 6.105, P < 0.001). MT and FL positively correlated to GMFM score (r > 0.391, P < 0.05), while SWV positively correlated to MAS score (r = 0.734, P < 0.001). ConclusionUltrasonography can be used to quantitatively evaluate the structure and stiffness of gastrocnemius muscle for children with spastic cerebral palsy.

This study aimed to elucidate the effect of tryptophan (Trp) on gut hormone secretion as well as the roles of the calcium-sensing receptor (CaSR) and its downstream signaling pathway in gut hormone secretion by assessing swine duodenal perfusion in vitro. Swine duodenum was perfused with Krebs-Henseleit buffer as a basal solution. Various concentrations (0, 10, and 20 mM) of Trp were applied to investigate its effect on gut hormone secretion. A CaSR antagonist was used to detect the involvement of CaSR and its signal molecules. The 20 mM Trp concentration promoted the secretion of cholecystokinin (CCK) and glucose-dependent insulinotropic peptide (GIP), elevated the mRNA level of CaSR, and upregulated the protein levels of CaSR, protein kinase C (PKC), and inositol trisphosphate receptor (IP3R). However, NPS 2143, an inhibitor of CaSR, attenuated the CCK and GIP release, reduced the mRNA level of CaSR, and decreased the protein levels of CaSR, PKC, and IP3R with 20 mM Trp perfusion. The results indicate that CCK and GIP secretion can be induced by Trp in swine duodenum in vitro, and the effect is mediated by CaSR and its downstream signal molecules PKC and IP3R.
Cholecystokinin ; Duodenum ; Gastric Inhibitory Polypeptide ; In Vitro Techniques ; Inositol ; Perfusion ; Protein Kinase C ; Receptors, Calcium-Sensing ; RNA, Messenger ; Swine ; Tryptophan

Objective A rat model of chronic post-thoracotomy pain is used to study whether acute pain and pre-operative diffused noxious inhibitory controls(DNIC) can predict chronic pain and how DNIC changes when pain maintains.Methods Rats were randomly divided into three groups:naive group,sham group and model group.DNIC was constantly assessed in individual rats,along with each animal's mechanical hyperalgesia and cold allodynia after thoracotomy.Results In model group,the incidence of chronic post-thoracotomy pain was 55%(11 of 20),which was named CPTP group,and the other 9 rats without chronic pain was defined as non-CPTP group.The pre-operative DNIC was significantly weaker in CPTP group with lower mechanical threshold on 6 days after surgery and higher cold sensitivity on 6 days after surgery comparing with non-CPTP group.In the acute pain phase (day 3),DNIC was decreased in both CPTP group and non-CPTP group as compared with pre-operative period.Besides,DNIC was recovered in non-CPTP group while kept impaired in CPTP group on 21 days after surgery.Conclusions Pre-operatively assessed DNIC efficiency and acute post-operative pain intensity were two independent predictors for CPTP.DNIC was decreased both in acute pain and chronic state,while returned to normal when pain sense was normal.

Objective To investigate the effect of dexamethasone combined with oridonin on proliferation and apoptosis in multiple myeloma cells U266 and the related molecular mechanism. Methods Exponential phase of growth U266 cells were treated with different concentrations of oridonin combined with dexamethasone or alone. U266 cells treated by DMSO were taken as control group. The proliferation inhibitory ratios were measured by CCK-8 assay followed by 24 h, 48 h and 72 h. Apoptosis induction was assessed by using Annexin V-FITC kit. Real time PCR was used to examine the mRNA changes of Notch1, NF-κB/p65 and bcl-2. Western blot assay was applied to detect the protein expression of Notch1, cleaved Notch1, NF-κB/p65 and bcl-2. Results Compared with that in control group, proliferation in all the experimental groups was inhibited (P<0.05), and the apoptosis was promoted (P<0.05); especially the combination of dexamethasone and oridonin had a synergistic effect on the proliferation and apoptosis of U266 cells (P<0.05). The results of PCR and Western blot showed that after treatment of U266 cells with dexamethasone, the mRNA as well as their protein levels of NF-κB/p65 and bcl-2 were decreased compared with those in the control group (P<0.05). Moreover, the mRNA and protein expression of Notch1, cleaved Notch1, NF-κB/p65 and bcl-2 was obviously down-regulated in oridonin group and the combination group (P<0.05). Conclusion Combination of dexamethasone and oridonin can significantly increase the anti-tumor effect by inhibiting proliferation and inducing apoptosis of U266 cells, which may be related to the inhibition of the Notch1 pathway.

Objective To investigate the application of contrast-enhanced ultrasonograhy CEUS in the early assessment of antiangiogenic treatment in metastatic colorectal cancer Methods Among 41 colorectal cancer patients with liver metastases enrolled in this study 25 patients received treatment of antiangiogenic treatment bevacizumab combined with chemotherapy BV group and 16 patients received chemotherapy only non-BV group To assess the treatment responses in the liver lesions between and within these two groups CEUS scans were performed before the treatment T0 or??baseline on the day 2 T1 and on the day 7 T2 after the treatment The percentage differences were evaluated according to the CEUS quantitative parameters such as rise time RT peak intensity PI mean transit time MTT area under the curve AUC time from peak to one half TPH wash in slope WIS and time to peak TTP Results On T1 both the RT and MTT increased within the BV group and decrease within the non-BV group when compared with their own baselines the differences were statistically significant P <0 05 On T2 the MTT increased within the BV group and decreased within the non-BV group when compared with their baselines the differences again were statistically significant P <0 05 All other parameters measured on T1 and T2 did not show a significant difference when compared with baselines Within the BV group the RT decreased and WIS and TPH increased for the responding subjects when compared with the non-responding subjects the differences were statistically significant P < 0 05 Within the non-BV group however no statistically significant differences were found for all parameters between the responding subjects and non-responding subjects P >0 05 Conclusions CEUS can be used to assess the treatment response in liver metastatic patients with colorectal cancer by monitoring the change of the tumor perfusion before and after the treatment.

Objective To detect serum oxytocin and highly sensitive C-reactive protein (hs-CRP) levels in obese and type 2 diabetes mellitus(T2DM) subjects and investigate the relationships between serum oxytocin levels and hs-CRP, glycolipid metabolism, insulin resistance and pancreas β cell function. Methods A total of 176 subjects were enrolled in the study, including 88 patients with newly-diagnosed type 2 diabetes ( T2DM) and 88 subjects with normal glucose tolerance(NGT). NGT and T2DM groups were further divided each into normal weight (NW) and obese(OB) subgroups. Obesity was defined as body mass index(BMI)≥25 kg/ m2 according to the WHO-Western Pacific Region diagnostic criteria (2000). 75g oral glucose tolerance test ( OGTT) was performed in all subjects. Fasting plasma glucose ( FPG), 2 h postprandial plasma glucose (2hPG), fasting insulin ( FINS), 2h postprandial serum insulin(2hINS), HbA1C and lipids were also determined. Insulin resistance and pancreas β-cell function were determined by homeostasis model assessment ( HOMA-IR, HOMA-β). Highly sensitive C-reactive protein(hs-CRP) level was determined by chemiluminescence immunoassay and fasting serum oxytocin level was determined by ELISA. Results Serum oxytocin level was lower in T2DM group than that in NGT group(P<0. 01), while serum hs-CRP level was higher in T2DM group than that in NGT group(P<0. 01). The level of serum oxytocin in subjects with obesity was also lower than that in subjects with NW in both NGT and T2DM groups [7. 16(6. 45-8. 82) vs 7. 98(7. 03-9. 17) ng/ L and 9. 23(8. 16-10. 36) vs 9. 86(8. 77-12. 06) ng/ L, P<0. 05]. The level of serum hs-CRP in subjects with obesity was higher than that in subjects with NW in both NGT and T2DM groups [0. 99(0. 25-1. 97) vs 0. 54(0. 19-0. 91) mg/ L and 3. 47(1. 63-6. 20) vs 1. 65(0. 81-3. 81) mg/ L, P<0. 05]. Serum oxytocin level was negatively correlated with hs-CRP, BMI, WC, WHR, HbA1C , FPG, 2hPG, FINS, 2hINS, total cholesterol, triglycerides, LDL-C and HOMA-IR, while was positively correlated with HOMA-β(P<0. 05). Subjects within the upper serum hs-CRP tertile had lower level of oxytocin when compared to subjects in the middle or lower serum hs-CRP tertiles(P<0. 05 ). Conclusion Serum oxytocin level was decreased in subjects with type 2 diabetes as well as with obesity. Serum oxytocin level was closely correlated with inflammation, glycolipid metabolism, insulin resistance, and pancreas β cell function. It may play an important role in the pathogenesis of obesity and T2DM.

Inferior vena caval (IVC) tumor thrombus with cardiac extension is a very rare phenomenon,which proliferates fast and could be very challenging to the surgery.This paper was designed to investigate the clinical value of transesophageal echocardiography (TEE) for the surgical resection of IVC tumor thrombus extending into right cardiac cavities.Six cases from our medical institute,preoperatively diagnosed as IVC tumor thrombus with cardiac extension and scheduled for the surgical resection,were retrospectively analyzed.In addition to real-time and dynamic monitoring,comprehensive TEE exams were performed for all the patients respectively after anesthesia induction,namely before tumor resection and after tumor resection.Cardiac extension was defined by the preoperative finding of cardiac mass originated from IVC tumor thrombus by transthoracic echocardiography,computerized tomography or CT angiography.In all the cases,intraoperative TEE provided an accurate and excellent view of the IVC tumor thrombus.For case three,the IVC tumor thrombus was found at the IVC entrance to right atrium without further cardiac extension; for case five,the IVC tumor thrombus proliferated into right atrium but the extended cardiac mass was very slim and flexible and the tricuspid valve was untouched; for case four,the IVC tumor thrombus extended into right atrium and even cross the tricuspid valve but the extended cardiac mass was also very slim and flexible.Based on the TEE-provided information,the originally scheduled surgical decision was modified and the surgical resection was performed without cardiopulmonary bypass (CPB).For the other three cases,the intraoperative TEE showed similar results to preoperative findings.The huge IVC tumor thrombus extended into the right heart,presented almost no flexibility and dramatically compromised the intracardiac blood flow.For the three cases,CPB support was indispensable for the tumor resection.The full TEE exam after tumor resection in all the six patients displayed clear surgical resection without tumor residuals,but in those three patients suffered with severely compromised cardiac extension,severe tricuspid regurgitation was noticed.All the six patients were closely monitored until discharged,and no TEE-related complications were observed.This paper reports about TEE' s utilization in a series of consecutive patients undergoing surgical resection of IVC tumor thrombus with cardiac extension.In addition to its safety and effectiveness,TEE can provide valuable information for surgical decision making,surgical intervention assessment and anesthesiamanagement strategies.

BACKGROUND:It is reported that the cellsurface antigen may be damaged by the trypsin enzyme in some extent and the cellactivity may also be influenced, as a result of which, the subsequent separation and fol ow-up-test wil be affected. Accutase enzymes possess the activities of protease and col agen enzyme and need no special termination or cleaning at the end of digestion. Another special function of accutase enzyme is to protect cellsurface antigen and thus it has been widely applied in the stem celldigestion and culture.
OBJECTIVE:To compare the digestive effect of accutase enzymes and trypsin enzyme in the separation and original culture of spermatogonial stem cells.
METHODS:The testes of 5-7 days male Kunming mice (n=45) were col ected and primarily digested with col agenas. The suspension of digested tissue was divided into three parts with the same volume named accutases enzyme group, trypsin enzyme group and mixed enzyme group (trypsin enzyme and hyaluronidase). For the comparison of testis digestive status and the time required for the formation of single cells, the micrographs were taken at 1, 3 and 5 minutes respectively after enzyme digestion. The total number and the mortality of the single cells were estimated and compared. The leydig cells and sertoli cells were removed by differential adherent method and the remained spermatogenic cells were then treated with CD90.2 immune magnetic beads. The selected spermatogonial stem cells were subsequently labeled by glial cellline-derived neurotrophic factor receptor alpha-1 and sorted with flow cytometry. The number of spermatogonial stem cells positive for glial cellline-derived neurotrophic factor receptor alpha-1 in CD90.2+and CD90.2-cells was compared within different groups.
RESULTS AND CONCLUSION:The digestive capacities of different enzyme were different. Accutases enzyme obtained the single cellsuspension more quickly than trypsin enzyme and mixed enzyme, and had the least cellmasses and broken cellmembrane than the other two groups. After the differential attached treatment, the highest total number of CD90+spermatogonial stem cells and lowest cellmortality could be found in the accutases group, when compared with the other groups. The results of cellsorting by flow cytometry showed a higher rate (72.24%) of GFRα1+/CD90+cells in the accutases group than the trypsin group (51.16%) and mixed enzyme group (71.27%). The GFRα1+/CD90-number in the accutases group was lower (15.03%) than that of the trypsin group (18.8%) and mixed enzyme group (24.23%), respectively. The results indicate a better effect of accutases enzyme on the primary separation of spermatogonial stem cells than that of trypsin or mixed enzyme.

Dietary essential amino acids are extracted in the first-pass metabolism by the small intestine.Besides incorporation into mucosal protein,they are also the fuels for the utilization by small intestinal mucosal cells and the precursors for intestinal synthesis of new amino acids and enormous biological important molecules such as glutathione,polyamines,etc.Thus,their gut metabolism is not only crucial for the maintenance of the integrity and function of intestinal mucosa,but also important for the regulation of whole body metabolism.