ObjectiveTo evaluate the clinical efficacy and safety of Tongluo Mingmu capsules in the treatment of diabetic retinopathy with blood stasis， collateral obstruction， and Qi and Yin deficiency syndrome. MethodA randomized， double-blind， positive-control， and multi-center clinical trial design method was used. 416 patients with diabetic retinopathy with blood stasis， collateral obstruction， and Qi and Yin deficiency syndrome in four test centers were included （the ratio of the treatment group to the control group was 3∶1）. On the basis of standardized hypoglycemic treatment， the treatment group was given both four Tongluo Mingmu capsules and two Calcium Dobesilate capsule agents three times a day， while the control group were given both two Calcium Dobesilate capsules and four Tongluo Mingmu capsule agents three times a day. The course of treatment was 12 weeks. The curative effect of Tongluo Mingmu capsules was evaluated by comparing the comprehensive curative effect of diabetic retinopathy， traditional Chinese medicine（TCM） syndrome score， corrected visual acuity， fundus changes， fundus fluorescence angiography， and other curative effect indexes before and after treatment in the two groups. At the same time， general examination， laboratory examination， and adverse events were performed to evaluate the safety of the drug. ResultThe baseline demographic data and disease characteristics of the treatment group and the control group were balanced and comparable， with the difference not statistically significant. After 12 weeks of treatment， the total effective rate of the comprehensive curative effect of diabetic retinopathy in the treatment group （61.0%， 189/310） was better than that in the control group （44.1%， 45/102）， and the difference was statistically significant （χ²=8.880， P<0.01）. The total effective rate of TCM syndromes in the treatment group （88.4%， 259/293） was better than that in the control group （69.9%， 65/93）， and the difference was statistically significant （χ²=17.927， P<0.01）. The disappearance rate of dry eyes （χ²=8.305）， dull complexion （χ²=4.053）， lassitude （χ²=10.267）， shortness of breath （χ²=8.494）， and dry stool （χ²=8.657） in the treatment group was higher than that in the control group， and the difference between the groups was statistically significant （P<0.05， P<0.01）. In terms of improving corrected visual acuity （χ²=8.382）， fundus changes （χ²=6.026） ， the treatment group was significantly better than the control group （P<0.05）. During the trial， the incidence of adverse events in the treatment group and the control group was 1.3% and 2.9%， respectively. There was no significant difference between the two groups. In addition， there were no serious adverse events and adverse events leading to withdrawal in both groups. ConclusionTongluo Mingmu capsules can improve the comprehensive curative effect of diabetic retinopathy and enhance the efficacy of TCM syndromes， visual acuity， fundus changes， and fundus fluorescein angiography， with great safety. Therefore， it can provide a new alternative therapeutic drug for patients with diabetic retinopathy.

Objective To prepare silymarin phospholipids complex（SM-PC） and investigate its physicochemical properties. Methods On the basis of single-factor tests, the drug-lipid ratio, drug concentration and reaction temperature were selected as the factors of the central composite design and response surface methodology in the preparation of SM-PC by solvent volatilization, and the best process was optimized with the compound rate as the index. And its in vitro dissolution was measured. Results The optimum preparation technology of SM-PC was as follows: acetone was used as compound solvent, the concentration of SM was 8.0 mg/ml, the mass ratio of SM to phospholipid was 1∶1.8, the reaction temperature was 56 ℃ and the recombination rate was（95.15±1.55）% with deviation of less than 3%. The in vitro dissolution test showed that the dissolution of SM-PC was close to 90% in 60 min. The dissolution behavior of main component of silybin was similar to that of silymarin capsules（Legalon ^®）, which was higher than SM-API. Conclusion SM-PC was successfully prepared by central composite design response surface method, which significantly improved the dissolution and laid a foundation for the study of subsequent preparations.

Objective To evaluate the release characteristics in vitro, pharmacokinetics in rabbits and in vivo-in vitro correlation of silymarin phospholipid complex microporous osmotic pump controlled release tablets（SM-PC MPOP）. Methods The release characteristics of SM-PC MPOP in vitro were detected by HPLC in the artificial gastric fluid. Six beagle dogs were subjected to double cycle cross control, which were given SM-PC MPOP and Legalon（30 mg/kg）. The concentration of silybin in plasma was determined by HPLC and the data were processed by software. Results The cumulative release rate of SM-PC MPOP in vitro was over 85% in 12 h. The pharmacokinetics in beagle dogs showed that SM-PC MPOP and legalon conformed to double compartment first-order absorption model and the pharmacokinetic parameters were obtained: t_max:（3.2±0.4）and（0.9±0.1）h, C_max:（0.298 6±0.068 9）and（0.629 9±0.076 5）μg/ml, AUC_0→24:（2.996 8±0.583 3）and（2.268 9±0.432 8）h·μg /ml. The relative bioavailability of SM-PC MPOP was（162.21 ± 30.82）%. Conclusion SM-PC MPOP could release slowly, which could increase the relative bioavailability significantly. The correlation between the absorption in vivo and release in vitro was fine（r = 0.839 0）.

Objective:To investigate the characteristics and risk factors of local recurrence in resected pancreatic cancer.Methods:The clinicopathological data of 190 patients in whom recurrent sites can be identified after radical resection of pancreatic cancer from Sep 2013 to Aug 2020 at the Cangzhou Central Hospital were retrospectively analyzed. The survival time and clinicopathological characteristics of local recurrence were compared with those of other recurrence types. Cox risk regression model was used to analyze the risk factors of local recurrence.Results:The recurrence sites were local (49 cases, 25.8%), liver (53 cases, 27.9%), lung (35 cases, 18.4%), peritoneal (25 cases, 13.2%) and multiple sites (28 cases, 14.7%). Patients mRFS and mOS were 17.8 months and 30.9 months respectively. The clinicopathological features of patients with local recurrence were compared with those of other recurrence types [tumor diameter ( P=0.023), preoperative CA199 level ( P=0.021), peripancreatic nerve plexus invasion ( P=0.031), lymphovascular invasion ( P=0.004), surgical margin state ( P<0.001) and postoperative adjuvant chemotherapy ( P=0.038)]. Tumor diameter ( P=0.018), peripancreatic nerve plexus invasion ( P=0.002) and postoperative adjuvant chemotherapy ( P=0.004) were independent factors for local recurrence in resected pancreatic cancer, and only peripancreatic nerve plexus invasion was not associated with other recurrence types. Conclusions:Local recurrence in resected pancreatic cancer has important impact on the prognosis of patients. Peripancreatic nerve plexus invasion is an independent factor affecting local recurrence.

Objective:To explore the role and mechanism of long non-coding RNA KCNQ1 overlapping transcript 1 (LncRNA KCNQ1OT1) in the migration, proliferation and invasion of hepatocellular carcinoma (HCC).Methods:The experimental method was conducted. The expression levels of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues in the StarBase database were collected. The experimental methods including real-time quantitative PCR, cell transfection, scratch assay, CCK8 assay, Transwell assay, Western blot were used to determine the expression, migration, proliferation, invasion of LncRNA KCNQ1OT1 in HCC cells and its relationship with phosphatidylinositol 3-kinase/phosphorylated AKT Protein (PI3K /p-AKT) signaling pathways. Observation indicators: (1) expression of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues; (2) the migration of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown; (3) the proliferation and invasion of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown; (4) effects of LncRNA KCNQ1OT1 gene knockdown on PI3K/p-AKT signaling pathways. Measurement data with normal distribution were expressed as Mean± SD, and comparison between groups was analyzed using the t test. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. Results:(1) Expression of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues. The expression levels of LncRNA KCNQ1OT1 in 374 HCC tissues and 50 normal liver tissues from StarBase database were 3.320±0.017 and 1.470±0.025, respectively, showing a significant difference ( t=5.24, P<0.05). Results of gene expression profile interactive analysis showed that the 30-month disease-free survival rates of HCC patients with high and low expression levels of LncRNA KCNQ1OT1 were 41% and 55%, respectively, with a significant difference ( χ2=6.209, P<0.05). The relative expression of LncRNA KCNQ1OT1 in HepG2, SMCC-7721and MHCC-97H cells were 1.470±0.042, 3.300±0.032, 4.040±0.031, respectively, versus 1.000±0.022 in normal liver cells (LO2), showing significant differences ( t=17.66, 95.40, 114.20, P<0.05). (2) The migration of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown. ① Results of cell transfection showed that the relative expression levels of LncRNA KCNQ1OT1 in HepG2, SMCC-7721 and MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.350±0.016, 0.310±0.020, 0.380±0.018, respectively, versus 1.000±0.021, 1.000±0.018, 1.000±0.019 in the negative control cells, showing significant differences ( t=23.40, 28.15, 22.32, P<0.05). ② Results of scratch assay showed that the healing rates of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 85.0%±1.9%, 75.0%±1.8%, 90.0%±1.7%, respectively, versus 100.0%±2.0%, 95.0%±1.8%, 72.0%±1.7% of the negative control cells, showing significant differences ( t=31.35, 47.36, 38.42, P<0.05). ③ Results of Transwell assay showed that the vertical migration rates of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 195±10, 205±12, 85±8, respectively, versus 520±11, 430±7, 405±20 of the negative control cells, showing significant differences between them ( t=922.30, 458.20, 708.40, P<0.05). (3) The proliferation and invasion of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown. ① Results of CCK8 assay showed that 72-hour optical densities of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 1.370±0.018, 1.240±0.016, 1.360±0.020, respectively, versus 0.900±0.023, 1.740±0.032, 1.230±0.025 of the negative control cells, with significant differences ( t=10.79, 12.00, 7.56, P<0.05). ② Results of Transwell assay showed that the invasion numbers of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 186±12, 155±7, 75±9, respectively, versus 505±1, 245±8, 300±15 of the negative control cells, showing significant differences ( t=955.90, 163.40, 530.90, P<0.05). (4) Effects of LncRNA KCNQ1OT1 gene knockdown on PI3K/p-AKT signaling pathways. Resluts of Western blot showed that the relative repression levels of PI3K in HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.447±0.009, 0.430±0.012, 0.354±0.006, respectively, versus 0.820±0.017, 0.850±0.012, 0.531±0.001 of the negative control cells, showing significant differences ( t=18.94, 25.72, 27.46, P<0.05). The relative repression levels of p-AKT in HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.343±0.015, 0.410±0.012, 0.579±0.006, respectively, versus 0.546±0.012, 0.620±0.012, 0.830±0.012 of the negative control cells, showing significant differences ( t=10.78, 12.86, 19.02, P<0.05). Conclusions:LncRNA KCNQ1OT1 plays an important role in the occurrence and development of HCC. LncRNA KCNQ1OT1 gene knockdown can inhibit the PI3K/AKT signaling pathways, so it can significantly inhibit the proliferation, migration and invasion of HCC cells.

Objective To compare the effects of cleaning oral among different nursing methods for patients with dysphagia after brain injury who take food by gastric tube as well as by mouth. Methods A total of 59 patients with dysphagia after brain injury were divided into research group ( 29 cases, mouth nursing by modified method of brushing their teeth and irrigation ) and control group ( 30 cases, mouth nursing by traditional oral care routine) according to the random number table method from October 2012 to May 2014. Results were observed by comparing oral condition, indexes of pharyngeal swab culture and water-swallowing test. Results Incidence rates of halitosis and unclean mouth were all 13. 8% in the research group which were lower than those (73. 3%, 63. 3%) in the control group with statistically significant differences (χ2 =21. 208, 15. 214;P<0. 01). The effective rate of water-swallowing test was 79. 3% in the research group which was higher than that (56. 7%) in the control group (χ2 =12. 202,P<0. 01). There was no statistically significant difference in abnormity rates between two groups before intervention (P >0. 05). The abnormity rate in the research group was lower than that in the control group at 4th week (χ2 =8. 958,P <0. 01). There was statistically significant difference in abnormity rate before interventions and at 4th week in the research group (P<0.01), with no statistically significant difference in the control group (P>0.05). The rate of oral mucosa bleeding was 3. 4% in the research group and 6. 7% in the control with no statistically significant difference (χ2 =0. 316,P >0. 05), but there was no mechanical injury of oral mucosa in two groups. There was no statistically significant difference in index of water-swallowing test between two groups before interventions ( P>0. 05), with significant difference at 4th week (P<0. 01). Conclusions The implementation of mouth nursing by modified method of brushing their teeth and irrigation not only can achieve the purpose to clean oral effectively, but also can reduce oral bacteria and promote the recovery of swallowing function.

Objective To evaluate the clinical value of the real-time three-dimensional ultrasound in the diagnosis of the fistula-in-ano.Methods One hundred and eighteen fistula-in-ano patients were examined using conventional and three-dimensional ultrasound,the stereotaxis of interal opening and the shape of the fistula-in-ano were analyzed.Then the diagnosis results of ultrasound were compared with the surgery and pathology.Results In 1 1 8 patients,the accuracy of preoperative identifying internal opening by two-dimensional ultrasound was 85%,and 95% for three-dimensional ultrasound with statistically significant difference (χ2 =6.679,P <0.05).For complex anal fistula,the diagnostic accuracy rates of main fistula tract by three-dimensional ultrasound (100%,82/82 )was higher than that by two-dimensional ultrasound(95%,78/82),the difference was statistically significant (χ2 =4.100,P <0.05 ).In 82 cases of complex anal fistula,the diagnostic accuracy of branch fistula tract by two-dimensional ultrasound was 70%(57/82 ),the accuracy using three-dimensional ultrasound was 89% (73/82 ),there was statistically significance (χ2 =9.499,P < 0.05 ).Conclusions Three-dimensional ultrasound can accurately locate the interal opening of fistula-in-ano and determine the shape of fistula-in-ano,which provided the most intuitive information for clinical treatment and had high practical value.

OBJECTIVETo report on clinical, genetic and molecular characterization of two Chinese families with Leber's hereditary optic neuropathy.

METHODSOphthalmological examinations have revealed variable severity and age at onset of visual loss among the probands and other matrilineal relatives of both families. The entire mitochondrial genome of the two probands was amplified with PCR in 24 overlapping fragments using sets of oligonucleotide primers.

RESULTSThe ophthalmological examinations showed that penetrance was 12.5% and 30.0% respectively in the two families. Sequence analysis of the complete mitochondrial genomes in these pedigrees has identified unreported homoplasmic T8821G mutation in the ATPase 6 gene and distinct sets of polymorphisms belonging to haplogroups M10a. The T8821G mutation has occurred at the extremely conserved nucleotide (conventional position 99) of the ATPase6. Thus, this mutation may alter structural formation of ATPase6, thereby leading to failure in the synthesis of ATP involved in visual impairment.

CONCLUSIONAbove observations have suggested that the ATPase6 T8821G mutation may be involved in the pathogenesis of optic neuropathy in these families.

Adolescent ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Male ; Mitochondrial Proton-Translocating ATPases ; genetics ; Molecular Sequence Data ; Optic Atrophy, Hereditary, Leber ; enzymology ; genetics ; Pedigree ; Point Mutation ; Young Adult

Objective To prepare insulin thiolated hyaluronic acid nanoparticles (Ins-HA-Cys-NPs)and study its physicochemical properties. Methods The Ins-HA-Cys-NPs was prepared by ultrasonic emulsifying method,and the properties of nanoparticles including morphology,mean diameter,Zeta potential,entrapment efficiency and drug loading efficiency were studied,as well as the cryoprotectant selection.Results The prepared nanoparticles was round in appearance and the mean diameter was(178.5 ±0.8)nm,the polydispersity index was (0.214 ±0.013)and the Zeta potential was -(38.47 ±0.46 )mV,while the entrapment efficiency was (48.85 ±0.66 )%,drug loading efficiency was (4.79 ±0.13 )%;10%mannitol as cryoprotectant provided uniform and well dispersed suspension of nanoparticles with blue opalescence after redispersion.Conclusion The thiolated hyaluronic acid nanoparticles may be used as the carrier for oral drug delivery system of insulin,and it provides a basis for studies on rats in vivo.

Objective To systematically review the efficacy of glutamine enriched nutrition support for patients with severe acute pancreatitis (SAP).Methods Database including PubMed,Embase,HighWire,Cochrane Central Register of Controlled Trials,Wanfang Database,CJFD and CBM were searched with glutamine,severe acute pancreatitis,SAP,谷氨酰胺,重症急性胰腺炎.Literatures published before March 2014 were searched.Randomized controlled trials containing the comparison of conventional treatment and glutamine enriched nutrition support were enrolled in the study,and then the literatures were screened and the data were extracted by 2 independent reviewers.The quality of the literatures was assessed,and the data were analyzed using the RevMan 5.2 software.SAP was diagnosed according to The revised Atlanta classificantion for acute pancreatitis or guideline for the diagnosis and treatment of SAP which was composed by the pancreatic surgery Branch of Chinese Medical Association.The count data were analyzed using the relative risk (RR) and 95％ confidence interval (95％ CI),and the measurement data were analyzed using standard mean difference (SMD) and 95 ％ CI.The heterogeneity of the data was analyzed using the I2 test.Results Ten literatures including 433 cases were enrolled in the study,and all of them were prospective randomized controlled studies.There were 218 patients treated by conventional methods (control group) and 215 patients received glutamine enriched nutrition support (experimental group).Compared with the control group,glutamine enriched nutrition support could elevate the albumin level,decrease the C-reaction protein level and shorten the hospital stay in the experimental group (SMD=1.00,-0.93,-0.71,95％CI:0.50-1.50,-1.25--0.61,-1.10--0.32,P＜0.05),glutamine enriched nutrition support could decrease the morbidity and mortality (RR =0.56,0.34,95％ CI:0.41-0.77,0.15-0.76,P ＜ 0.05) without increasing the expenses (SMD =0.03,95％ CI:-0.88-0.95,P ＞ 0.05).Conclusion Glutamine enriched nutrition support is superior to conventional methods for the treatment of SAP.