Objective Targeted adenoviral gene delivery from peripheral nerves was used to integrally analyse the characterization and time course of LacZ gene (AdLacZ) retrograde transfer to spinal cord and transgene product anterograde labeling ofperipheral nerve. Methods Recombinant replication-defective adenovirus containing AdLacZ was administrated to the cut proximal stumps of median and tibial nerves in Wister rats. Then the transected nerve was repaired with 10-0 nylon sutures. At different time point postinfection the spinal cords of C5 to T1 attached with DRGs and brachial plexuses, or L2 to L6 attached with DRGs and lumbosacralplexuses were removed. The removed spinal cord and DRGs were cut into 50 μm serialcoronal sections and processed for X-gal staining and immunohistochemical staining. The whole specimens of brachial or lumbosacral plexuses attaching with theirperipheral nerves were processed for X-gal staining. The number of X-gal stained neurons was counted and the initial detected time of retrograde labeling, peaktime and persisting period of gene expression in DRG sensory neurons, spinal cord motor neurons and peripheral nerves were studied. Results The gene transfer was specifically targeted to the particular segments of spinal cord andDRGs, and transgene expression was strictly unilaterally corresponding to the infected nerves. Within the same nerve models, the initial detected time of gene expression was earliest in DRG neurons, then in the motor neurons and latest in peripheral nerves. The persisting duration of β-gal staining was shortest in motor neurons, then in sensory neurons and longest in peripheral nerves. The initial detected time of β-gal staining in median nerve models was earlier in mediannerve models compared with that in the tibial nerve models. Although the initial detected time and the beginning of peak duration of β-gal staining were not same, the decreasing time of β-gal staining in motor and sensory neurons of thetwo nerve models were started at about the same day 8 post-infection. The labeled neurons were more in tibial nerve-models than that in median nerve models. Within the same models, the labeled sensory neurons of DRGs were morethan labeled motor neurons of ventral horn. The β-gal staining was tenser in median nerves than that in tibial nerves. However the persisting time of β-gal staining was longer in tibial nerve models. Conclusion The b gene expression in neurons and PNS renders this system particularly attractive for neuroanatomical tracing studies. Furthermore this gene delivery method allowing specific targeting of motor and sensory neurons without damaging the spinal cord might offer potentialities for the gene therapy of peripheral nerve injury.
Objective To explore the expression and function of NDRG2 gene in human primary hepatocellular carcinoma and normal hepatic tissues. Methods The immunohistochemical ABC method， Western blot， and Real-time PCR were used to investigate the expression and content of NDRG2 in human hepatocellular carcinoma and hepatic normal biopsies. Results The NDRG2 protein located in cytoplasm. The positive rate was 16.67%（5/30） and 100%（30/30） in hepatocellular carcinoma and normal hepatic tissues, respectively. The relative content of NDRG2 protein in hepatocellular carcinoma and normal hepatic tissues were 0.029 0±0.005 9 and 0.109 2±0.002 8. There were significant differences between human hepatocellular carcinoma and hepatic normal biopsies both in staining positive rates and relative content（P＜0.05）. The Western blot also agreed with the result，the expression level of NDRG2 protein in hepatocellular carcinoma and normal hepatic tissues was 1.13±0.15 and 1.57±0.18， respectively, there was significant difference（P＜0.05）. Also, compared with normal hepatic tissues, the expression level of NDRG2 mRNA in carcinoma tissues was reduced significantly (0.89±0.15 vs. 1.48±0.17, P＜0.05). However, there were no significant differences in NDRG2mRNA expression between Edmondson-Steiner grades. Conclusions There possibly have difference in NDRG2 expression between human primary hepatocellular carcinoma and normal hepatic tissue. NDRG2 gene may take part in the pathogenesis of human primary hepatocellular carcinoma. Futher study will be needed to study its mechanism and function.
ObjectiveTo observe the effects of NDRG1 on proliferation, migration and lumen formation of retinal vascular endothelial cells (RF/6A cells) in monkeys under high glucose condition. MethodsRF/6A cells were divided into normal group, mannitol group, high glucose group, small interfering RNA (siRNA) negative control group without target gene (siRNA group), 30 nmol/L siRNA down-regulated NDRG1 genome (siNDRG1 group) and 50 nmol/L siNDRG1 group. Normal group cells were cultured conventionally. The mannitol group was added with 25 mmol/L mannitol, and the high-glucose group was added with 25 mmol/L glucose. In the siRNA group, 25 mmol/L glucose was added, and then blank siRNA was added for induction. The 30 and 50 nmol/L siNDRG1 groups were added with 25 mmol/L glucose and induced with 30 and 50 nmol/L siRNDRG1, respectively. All cells were incubated for 24 h for follow-up experiments. Cell proliferation was observed by 4', 6-diaminidine 2-phenylindole staining. Cell counting kit-8 staining was used to detect cell activity. The expression level of NDRG1 mRNA and protein was detected by Western blot and real-time quantitative polymerase chain reaction. Cell migration was observed by cell scratch assay. Cell lumen formation assay was used to detect lumen formation. The two-tailed Student t test was used to compare the two groups. One-way analysis of variance was used to compare groups. ResultsThere were significant differences in cell proliferation rate (t=36.659, 57.645) mobility rate (t=24.745, 33.638) and lumen formation number (t=41.276, 22.867) between high glucose group and normal group and mannitol group (P＜0.01). Compared with normal group and mannitol group, the relative expression levels of NDRG1gene mRNA and protein in high glucose group were significantly decreased, with statistical significance (t=46.145, 21.541, 36.738, 32.976; P＜0.001). Compared with the siRNA negative group, the relative expression levels of NDRG1gene mRNA and protein in 30 nmol/L siNDRG1 group and 50 nmol/L siNDRG1 group were significantly decreased, and the differences were statistically significant (t=44.275, 40.7577, 57.167, 25.877; P＜0.01). Compared with normal group and siRNA group, cell mobility in 30 nmol/LsiNDRG1 group was increased, and the difference was statistically significant (t=57.562, 49.522; P＜0.01). Compared with normal group and siRNA group, the number of cell lumen formation in 30 nmol/LsiNDRG1 group was significantly increased in the same field of vision, and the difference was statistically significant (t=63.446, 42.742; P＜0.01). ConclusionDown-regulation of NDRG1 gene can improve the activity, migration and lumen formation of RF/6A cells under hyperglycemia.
Objective To systematically review the issues and countermeasures in supervising medical insurance funds under the DRG/DIP payment model. Methods The CNKI, WanFang Data, CBM, VIP, PubMed, Web of Science, and Embase databases were electronically searched to collect studies related to objectives from inception to March 15, 2024. Two reviewers independently screened the literature, extracted data, and assessed the quality of the included studies using the Critical Appraisal Skills Programme (CASP). Excel 2019 was used for data extraction and organization, and a thematic synthesis approach was employed for analysis. Results Nineteen qualitative studies were included. Nine studies identified key issues in fund supervision under the DRG/DIP model: inadequate regulatory mechanisms, weak regulatory capacity, low informatization, and traditional regulatory concepts. Fourteen studies proposed optimization strategies, including establishing a regulatory system, customizing regulatory indicators, creating a performance evaluation mechanism, developing talent, promoting multi-party regulation, enhancing intelligent supervision systems, and improving stakeholder collaboration and communication. Conclusion The DRG/DIP payment model faces challenges in medical insurance fund supervision, including weak mechanisms and capacity. Improving regulatory efficiency and ensuring medical service quality requires strengthening the regulatory system, customizing indicator systems, and enhancing talent development.
ObjectiveAs few studies have evaluated the policy effects of the Chinese simplified DRGs-PPS systematically, this research aims to assess its policy effects and to provide insight for other developing regions that are undergoing the same reform. MethodsThe history and major problems of the Chinese DRGs-PPS were analyzed qualitatively. Moreover, the efficiency (average hospitalization cost; length of stay, LOS) and equity of the simplified DRGs-PPS were examined at both macro and micro levels. ResultsAs of today, only 20 of the 32 provinces in mainland China had implemented the simplified DRGs. There were also huge differences in terms of the number and categories of diseases among the various provinces involved. Literature review showed that " lack of rationale in setting payment standards" , "limited diseases are included into the DRGs categories" and "lack of regulation to avoid ethical risks of health service providers" were the frequently cited problems. On the macro level, the national average medical cost had increased while the average LOS had been relatively stable from the year 2004 onwards, and simplified DRGs had been implemented widely since 2004 while discrepancies existed in various provinces. On the micro level, among the studies that focused on assessing hospitals with statistical test, 78% (11/14) of these studies revealed that hospitalization cost could be reduced and 60% (6/10) of them indicated that LOS could be reduced. ConclusionBy comparing the policy effects at both macro and micro levels, we conclude that the simplified DRGs are useful in controlling hospitalization cost but they fail to reduce LOS. Also much more still needs to be done in China to facilitate the transition from simplified DRGs to genuine DRGs.
Objective To perform data-driven, assisted prediction of health insurance reimbursement ratios for the major thoracic surgery group in CHS-DRG, in addition to providing an optional solution for health insurance providers and medical institutions to accurately and effectively predict the references of health insurance payments for the patient group. Methods Using the information on major thoracic surgery cases from a large tertiary hospital in Sichuan province in 2020 as a sample, 70% of the total dataset was used as a training dataset and 30% as a test dataset. This data was used to predict health insurance spending through a multiple linear regression model and an improved machine learning method that is based on feature selection. Results When the number of filtered features was the same via three machine learning methods including random forest, logistic regression, and support vector machine, there was no significant difference in the prediction effectiveness. The model with the best prediction effect had an accuracy of 78.96%, sensitivity of 83.93%, specificity of 71.27%, precision of 0.818 8, AUC value of 0.841 4, and a Kappa value of 0.610 8. Conclusion The basic characteristics such as the number of disease diagnoses and surgical operations, as well as the age of patients affect the reimbursement ratio. The cost of materials, drugs, and treatments has a greater impact on the reimbursement ratio. The combined method of feature selection and machine learning outperforms traditional statistical linear models. When dealing with a larger dataset that has many features, selecting the right number can enhance the prediction ability and efficiency of the model.