Objective:To investigate the effect of poly (lactic acid)(PLA)electrospun membranes loaded with cisplatin and chloroquine on the oral squamous cell carcinoma CAL-27 cells,and to explore the method to prevent the recurrence of oral cancer.Methods: The DDP/PLA membranes, CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were prepared by electrospinning.Then the micro morphology of three kinds of membranes were observed by scanning electron microscope (SEM);the degradation rate of PLA membrane was measuredby UV spectrophotometric.The LC3-Ⅱ expression level in CAL-27 cells was detected by laser scanning confocal microscope.The survival rate of CAL-27 cells was detected by MTT method.Results:The SEM results showed that the nanofibers of DDP/PLA,CQ/DDP/PLA and CQ/DDP/CQ/PLA membranes were continuous and smooth with uniform diameters.The degrated time of membranes was about 21 d.The MTT result showed that compared with control group,at first,the effects of cell killing of DDP/PLA membranes,CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were not obvious;as the extension of time,the survival rates of CAL-27 cells in DDP/PLA membranes group,CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were decreased (P <0.05).The immunofluorescence results showed that the fluorescence intensity of LC3-Ⅱ in CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were lower than that in DDP/PLA membranes group.Conclusion:CQ/DDP/PLA membranes with sustained-release effect can increase the sensitivity of CAL-27 cells to DDP and enchance the killer effect of DDP on the CAL-27 cells.

0.05).Conclusions The SCIO system is practical to assess the therapeutic effects of itraconazole and terbinafine for patients with onychomycosis.Treatment of onychomycosis with the two drugs is equally effective and safe.

Objective:To evaluate the effects of exogenous biliverdin (BV) on the expression of Litaf in PC12 cells subjected to oxygen-glucose deprivation and restoration (OGD/R).Methods:PC12 cells were seeded in a 96-well cell culture plate at a density of 1×10 4 cells/well for 3 days and were divided into 3 groups ( n=18 each) by a random number table method: control group (group C), OGD/R group, and biliverdin group (BV group). Group C was incubated in a 37 ℃ incubator (95% air+ 5%CO 2) for 6 h. To establish the OGD/R model, cells were incubated with sugar-free medium in a 37 ℃ incubator (95% air+ 5%CO 2) for 2 h, and the medium was then replaced with normal medium and cells were continuously incubated in a 37 ℃ incubator (95% N 2+ 5% CO 2). In BV group, 2 μg/ml biliverdin was added immediately after oxygen-glucose restoration.Cells in 6 wells in each group were selected at 6 h of restoration for determination of the expression of Litaf protein and mRNA (by real-time polymerase chain reaction) and tumor necrosis factor-alpha (TNF-α) concentration (by enzyme-linked immunosorbent assay). Results:Compared with group C, the expression of Litaf protein and mRNA was significantly up-regulated, and TNF-α concentration in supernatant was increased in group OGD/R ( P<0.05). Compared with group OGD/R, the expression of Litaf protein and mRNA was significantly down-regulated, and TNF-α concentration in supernatant was decreased in group BV ( P<0.05). Conclusion:The mechanism by which exogenous biliverdin reduces OGD/R damage to PC12 cells is related to inhibiting up-regulated expression of Litaf and alleviating the inflammatory responses.

Objective:To evaluate the effect of propofol anesthesia on autophagy in hippocampal neurons of newborn rats.Methods:Thirty-nine healthy Sprague-Dawley rats, aged 7 days, weighing 10-12 g, were divided into 3 groups ( n=13 each) using a random number table method: control group (group C), fat emulsion group (group F) and propofol group (group P). Normal saline 8 ml/kg was intraperitoneally injected for 5 consecutive days in group C. Medium-/long-chain fatty emulsion injection 8 ml/kg was intraperitoneally injected for 5 consecutive days in group F. Medium-/long-chain propofol injection 80 mg/kg was intraperitoneally injected for 5 consecutive days in group P. Five rats were sacrificed on 1st day after the end of propofol anesthesia, and hippocampal tissues were taken for determination of the expression of microtubule-associated protein 1 light chain 3B (LC3B) and Beclin-1 (by Western blot). The remaining rats in each group underwent the Morris water maze test on 19th day after the end of propofol anesthesia (30 days after birth), and the escape latency, percentage of time of staying at the target quadrant and the number of crossing the original platform were recorded. Results:Compared with group C, no significant change was found in the expression of hippocampal LC3B and Beclin-1, escape latency, percentage of time of staying at the target quadrant, and the number of crossing the original platform in group F ( P>0.05), and the expression of hippocampal LC3B and Beclin-1 was significantly up-regulated, the escape latency was prolonged, percentage of time of staying at the target quadrant was decreased, and the number of crossing the original platform was decreased in group P ( P<0.05 or 0.01). Conclusion:The mechanism by which propofol anesthesia causes long-term cognitive dysfunction may be related to promoting autophagy in hippocampal neurons of newborn rats.

Objective:To study the efficacy and safety of bone-filling mesh container plasty in the treatment of posterior wall fracture of vertebra caused by spinal metastases.Methods:This study is a retrospective analysis of 65 patients with pathological fractures of the vertebra caused by vertebral metastases treated with bone-filling mesh container plasty from January 2015 to December 2019. There were 21 males and 44 females, 70.3±10.8 (46-90) (years). According to primary tumor, there were 25 cases of lung cancer, 14 cases of breast cancer, 11 cases of digestive system cancer, 13 cases of urinary system cancer, 1 case of lymphoma and 1 case of ovarian cancer. In the segment of vertebral metastases, there were 2 cases of T 2 vertebra, 1 case of T 5 vertebra, 1 case of T 6 vertebra, 2 cases of T 8 vertebra, 1 case of T 9 vertebra, 5 cases of T 10 vertebra, 4 cases of T 11 vertebra, 15 cases of T 12 vertebra, 12 cases of L 1 vertebra, 8 cases of L 2 vertebra, 8 cases of L 3 vertebra, 4 cases of L 4 vertebra, and 2 cases of L 5 vertebra. According to the CT images of the patient's vertebra before operation, the area of the damaged posterior wall of the vertebra is measured as s, and the area of the posterior wall of the intact vertebra is measured as S. The ratio of posterior wall damage is calculated as R= s/ S, and the value of R represents the degree of damage to the posterior wall of the vertebra. According to the size of the R value, the patients were divided into four groups, typeⅠ( R≤25%, 21 cases), typeⅡ(25%< R≤50%, 22 cases), typeⅢ (50%< R≤75%, 14 cases), typeⅣ( R>75%, 8 cases). The visual analog scale (VAS), Oswestry disability index (ODI) and activity of daily living (ADL) before and 1 day after surgery, 1 month after surgery, and 3 months after surgery were analyzed and compared to evaluate the efficacy of bone-filling mesh container plasty. Pairwise comparisons were performed to verify whether there is a difference in efficacy, bone cement leakage and postoperative complications. Results:All 65 patients were followed up for 3-6 months, with an average of 3.8 months. The VAS scores before surgery, postoperative day 1, postoperative 1, 3 months were 7.32±0.99, 4.14±1.06, 4.11±0.97, and 4.34±1.11, respectively, with a statistically significant difference ( F=149.20, P<0.001). ODI of preoperative, postoperative day 1, postoperative 1, and 3 months were 69.45%±4.15%, 36.65%±3.72%, 36.84%± 3.38%, 37.78%±3.45%, respectively, with a statistically significant difference ( F=840.88, P<0.001). ADL score of preoperative, postoperative day 1, postoperative 1, and 3 months were 71.31±12.81, 79.85±9.14, 78.92±8.95, and 78.31±8.67, respectively, with a statistically significant difference ( F=149.20, P<0.001). There was no significant difference in VAS, ODI and ADL scores between types I and IV (all P>0.05), but with the increase of R value, the leakage rate of intraspinal bone cement would increase correspondingly. Eleven cases occurred bone cement leakage with the rate of 17%. The leakage rate of type I and II was 0, type III was 7.1% (1/14), and type IV was 37.5% (3/8). All patients did not have systemic complications such as allergies, shock, decreased oxygen saturation, etc., and there were no bleeding, infection, nerve root symptoms or cement insertion syndrome after surgery. Conclusion:Bone-filling mesh container plasty can significantly improve the pain symptoms of patients with spinal metastases and recovery functions. The degree of damage to the posterior vertebra has no effect on the efficacy of the surgery. As the degree of damage to the posterior wall of the vertebra increases, the risk of complications of bone cement leakage in the spinal canal will increase.

For the past few years, viral diseases such as Coronavirus Disease 2019, Influenza, and Ebola hemorrhagic fever have become more severe and more frequent. For people’s health and the stable operation of the national economy, it is of increasing importance to research the pathogenesis of virus and host immune regulation mechanism, analyze the regulation of gene expression between virus and host, and surveil pathogenic viruses and prevent virus diseases. Transcriptome sequencing is a kind of technology at the RNA level that focuses on the expression characteristics of genes in a specific space-time state, and it is an indispensable tool for analyzing differential gene expression and researching differential splicing of mRNA. With the progress of molecular biology experimental technology and the maturity of bioinformatics analysis platforms, transcriptomics is developing towards the low-cost and low technical threshold, which is gradually transitioning from the research field to the clinical laboratory and changing the traditional cognition of clinical staff in viruses and viral diseases. It has been increasingly used in the research of viral transcription mechanisms, immune interaction between virus and host, tracking disease progression, and antiviral drug development. In order to provide a methodological reference for the research of virus mechanism and prevention and control of virus disease, here, we mainly review meta-transcriptome from three aspects: technical process, common software and application scenarios of virus and virus diseases, and briefly mention the method and applications of other transcriptomes, including a population of cells RNA sequencing (Bulk RNA-seq), Single-cell RNA sequencing (ScRNA-seq), Single-nuclear RNA sequencing (SnRNA-seq), and Spatial Transcriptome (ST).

OBJECTIVETo compare the effects of acupuncture, electroacupuncture (EA) and moxibustion on functional constipation in rats.

METHODSSixty male Sprague-Dawley rats were divided into a control group (=8), a model group (=11), a medication group (=8), an acupuncture group (=11), an EA group (=11) and a moxibustion group (=11) by random number table. The rats in the model group, medication group, acupuncture group, EA group and moxibustion group were treated with intragastric administration of loperamide hydrochloride for 6 days continuously to establish the functional constipation models, while equal volume of drinking water was administrated to rats in the control group at the same time. The rats in the acupuncture group, EA group and moxibustion group were respectively treated with acupuncture, EA and moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) one hour after intragastric administration; rats in the medication group were treated with intragastric administration of cisapride suspension. All the treatment was given once a day for 6 days. At the last day of intervention, the 24-hour food intake, stool quantity and its water content were measured in each group; the pushing rate of intestine was measured; the structure of colon tissue and acidic mucus in its mucous layer were observed by hematoxylin-eosin dyeing and alcian blue dyeing; the expression of stem cell factor (SCF) and c-kit mRNA was detected by real-time PCR.

RESULTSCompared with the control group, the 24-hour food intake and stool quantity were reduced in the model group (both<0.01), and the water content of stool and pushing rate of intestine were reduced (both<0.01); compared with the model group, the stool quantity and its water content were increased in the medication group, acupuncture group, EA group (<0.05,<0.01), which were not significantly different from those in the moxibustion group (both>0.05). The pushing rate of intestine in each intervention group was increased (all<0.01). The 24-hour food intake and stool quantity in the medication group were not significantly different from those in the acupuncture group, EA group and moxibustion group (all>0.05), and the water content of stool was only reduced in the moxibustion group (<0.01). The pushing rate of intestine in the acupuncture group and moxibustion group was lower than that in the medication group (both<0.01), while that in the EA group was not significantly different from that in the medication group (>0.05). The water content of stool in the moxibustion group was lower than that in the acupuncture group and EA group (both<0.01). The pushing rate of intestine in the acupuncture group and moxibustion group was lower than that in the EA group (both<0.01). The HE staining result indicated the structure of colon tissue was normal, complete and similar in each group; the alcian blue staining indicated the acidic mucosubstance in the model group was lower than that in the control group; compared with the model group, the acidic mucosubstance in the medication group, acupuncture group, EA group and moxibustion group was all increased. Compared with the control group, the expression of SCF and c-kit mRNA was reduced in the model group (both<0.05); compared with the model group, the expression of SCF and c-kit mRNA was increased in the medication group, acupuncture group, EA group and moxibustion group (all<0.05); compared with the moxibustion group, the expression of c-kit mRNA was reduced in the acupuncture group and EA group (both<0.05).

CONCLUSIONSAcupuncture, EA and moxibustion all can play a positive regulative role on functional constipation in rats, in which EA has the best efficacy, followed by acupuncture.

ObjectiveTo investigate the mechanism of Tangbikang dry paste in the prevention and treatment of type 2 diabetic peripheral neuropathy （DPN） based on the glyoxalase-1 （GLO-1）/advanced glycation end products （AGE）/receptor for advanced glycation end products （RAGE） pathway. MethodsA total of 56 Sprague-Dawley rats were randomly divided， with eight assigned to the normal group. The remaining 48 rats were fed a high-fat diet combined with intraperitoneal injection of streptozotocin （STZ） to induce a type 2 diabetes mellitus （T2DM） model. Based on blood glucose levels， the rats were randomly assigned to the model group， Tanglin group （13.5 mg·kg^-1）， metformin group （135 mg·kg^-1）， and Tangbikang dry paste low-， medium-， and high-dose groups （3， 6， 12 g·kg^-1）. Successful modeling of DPN was confirmed by a decrease in mechanical pain threshold in the model group at week 4. Fasting blood glucose， body weight， and mechanical pain threshold were measured every 4 weeks. After 16 weeks of intervention， the pathological morphology of the sciatic nerve was observed using hematoxylin-eosin （HE） staining. The expression of RAGE， AGE， protein kinase C （PKC）， and collagen （COL） in the sciatic nerve was assessed by immunohistochemistry. The mRNA expression of RAGE， PKC， Toll-like receptor （TLR）， COL， and GLO-1 was detected using real-time quantitative PCR （Real-time PCR）. Serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， creatinine （CREA）， urea （UREA）， interleukin-6 （IL-6）， and tumor necrosis factor （TNF）-α were measured by enzyme-linked immunosorbent assay （ELISA）. ResultsCompared with the normal group， the model group showed significantly increased fasting blood glucose （P<0.01）， decreased body weight and mechanical pain threshold （P<0.01）， and elevated serum AST， ALT， CREA， UREA， IL-6， and TNF-α levels （P<0.01）. The expression of RAGE， AGE， and PKC in the sciatic nerve was significantly increased （P<0.01）， while COL expression was decreased （P<0.01）. The mRNA expression of TLR， RAGE， and PKC was upregulated （P<0.01）， whereas COL and GLO-1 mRNA levels were downregulated （P<0.01）. Histological examination showed irregular nerve morphology， axonal alterations， and myelin degeneration. Compared with the model group， fasting blood glucose levels in the Tangbikang dry paste high-dose group at all time points and in the medium-dose group at weeks 4 and 16 were significantly reduced （P<0.05， P<0.01）. No significant changes in body weight were observed across all Tangbikang dose groups. The mechanical pain threshold was elevated at different time points after administration in all Tangbikang groups （P<0.05， P<0.01）. Serum IL-6 and TNF-α levels were decreased in all dose groups （P<0.05， P<0.01）. The expression of RAGE， AGE， and PKC in the sciatic nerve was reduced （P<0.01）， while COL expression was increased （P<0.01）. The mRNA expression of TLR， RAGE， and PKC was downregulated （P<0.01）， whereas GLO-1 mRNA expression was upregulated （P<0.05， P<0.01）. Additionally， COL mRNA expression was significantly increased in the low- and high-dose groups （P<0.01）. Pathological changes in the sciatic nerve were milder in all Tangbikang groups compared to the model group. ConclusionTangbikang dry paste significantly improves DPN， and its mechanism may be associated with the regulation of the GLO-1/AGE/RAGE signaling pathway.

ObjectiveTo investigate the mechanism of Tangbikang dry paste in the prevention and treatment of type 2 diabetic peripheral neuropathy （DPN） based on the glyoxalase-1 （GLO-1）/advanced glycation end products （AGE）/receptor for advanced glycation end products （RAGE） pathway. MethodsA total of 56 Sprague-Dawley rats were randomly divided， with eight assigned to the normal group. The remaining 48 rats were fed a high-fat diet combined with intraperitoneal injection of streptozotocin （STZ） to induce a type 2 diabetes mellitus （T2DM） model. Based on blood glucose levels， the rats were randomly assigned to the model group， Tanglin group （13.5 mg·kg^-1）， metformin group （135 mg·kg^-1）， and Tangbikang dry paste low-， medium-， and high-dose groups （3， 6， 12 g·kg^-1）. Successful modeling of DPN was confirmed by a decrease in mechanical pain threshold in the model group at week 4. Fasting blood glucose， body weight， and mechanical pain threshold were measured every 4 weeks. After 16 weeks of intervention， the pathological morphology of the sciatic nerve was observed using hematoxylin-eosin （HE） staining. The expression of RAGE， AGE， protein kinase C （PKC）， and collagen （COL） in the sciatic nerve was assessed by immunohistochemistry. The mRNA expression of RAGE， PKC， Toll-like receptor （TLR）， COL， and GLO-1 was detected using real-time quantitative PCR （Real-time PCR）. Serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， creatinine （CREA）， urea （UREA）， interleukin-6 （IL-6）， and tumor necrosis factor （TNF）-α were measured by enzyme-linked immunosorbent assay （ELISA）. ResultsCompared with the normal group， the model group showed significantly increased fasting blood glucose （P<0.01）， decreased body weight and mechanical pain threshold （P<0.01）， and elevated serum AST， ALT， CREA， UREA， IL-6， and TNF-α levels （P<0.01）. The expression of RAGE， AGE， and PKC in the sciatic nerve was significantly increased （P<0.01）， while COL expression was decreased （P<0.01）. The mRNA expression of TLR， RAGE， and PKC was upregulated （P<0.01）， whereas COL and GLO-1 mRNA levels were downregulated （P<0.01）. Histological examination showed irregular nerve morphology， axonal alterations， and myelin degeneration. Compared with the model group， fasting blood glucose levels in the Tangbikang dry paste high-dose group at all time points and in the medium-dose group at weeks 4 and 16 were significantly reduced （P<0.05， P<0.01）. No significant changes in body weight were observed across all Tangbikang dose groups. The mechanical pain threshold was elevated at different time points after administration in all Tangbikang groups （P<0.05， P<0.01）. Serum IL-6 and TNF-α levels were decreased in all dose groups （P<0.05， P<0.01）. The expression of RAGE， AGE， and PKC in the sciatic nerve was reduced （P<0.01）， while COL expression was increased （P<0.01）. The mRNA expression of TLR， RAGE， and PKC was downregulated （P<0.01）， whereas GLO-1 mRNA expression was upregulated （P<0.05， P<0.01）. Additionally， COL mRNA expression was significantly increased in the low- and high-dose groups （P<0.01）. Pathological changes in the sciatic nerve were milder in all Tangbikang groups compared to the model group. ConclusionTangbikang dry paste significantly improves DPN， and its mechanism may be associated with the regulation of the GLO-1/AGE/RAGE signaling pathway.