OBJECTIVE:To establish the quality standard of Chushi pill. METHODS:Microscopic identification and TLC were adopted for the qualitative identification of Cortex moutan,C. dictamni,Angelica sinensis,Rubia cordifolia and Gardeniae fructus in Chushi pill;HPLC was performed to determine the contents of paeonol and baicalin. It was performed on column of Kro-masil 100-5 C18 with mobile phase of methanol-water-phosphoric acid(47∶53∶0.2,V/V/V)at the flow rate of 1.0 ml/min,the detec-tion wavelength was 280 nm,the temperature was 25 ℃ and the volume was 10 μl. RESULTS:The microscopic identification showed microscopic characteristics of C. moutan and C. dictamni,and characteristics of A. sinensis,R. cordifolia and G. fructus were identified by TLC;the linear range of paeonol was 0.106 24-2.124 8 μg(r=0.999 9)and baicalin was 0.059 04-1.180 8 μg (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 2.06%;average recoveries were respective-ly 101.56%(RSD=1.68%,n=9)and 100.16%(RSD=1.13%,n=9). CONCLUSIONS:The method is simple,accurate and re-producible,and can be used for the quantity control of Chushi pill.

To investigate the RNA interference (RNAi) effect induced by vector-derived small interfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online, we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene. Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different, depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80.

OBJECTIVETo study and establish the optimal technology for the preparation of evodiae juice.

METHODThe contents of evodiamine, rutaecarpine and evodin were simultaneously determined with HPLC, and each yield of the three compounds were chosen as the evaluating indicator. The orthogonal test coupled with the weighted coefficient method were adopted to acquire the optimal technology for the preparation of evodiae juice.

RESULTThe study showed that the optimal technology for the preparation of evodiae juice was as follows: decocted three times while the first time was with 12-fold of water socked 30 minutes and decocted 45 minutes, the second time was with 8-fold of water decocted 20 minutes and the third time was with 6-fold of water decocted 20 minutes.

CONCLUSIONThis method is simple and accurate. The optimal technology is suitable for industry manufacture of evodiae juice.

Objective To investigate the sleep quality of inland military officers and soldiers stationed and trained at plateau and its possible influence factors.Methods A total of 459 military officers and soldiers stationed and trained at plateau were performed the on site psychological assessment by adopting the psychological stress self-evaluation test (PSET),Pittsburgh Sleep Quality Index (PSQI),work related fatigue feelings (WRFFQ) and self compiled general data questionnaire.Results (1) The mean value of overall sleep quality in militaries stationed and trained at plateau was 5.61-±-3.48.40.5 ％ of respondents had good sleep quality,25.3％ had poor sleep quality and 34.2％ had general sleep quality.(2)The fatigue and psychological stress scores in the militaries with poor sleep quality were significantly higher than those in the militaries with good sleep quality (t1 =10.70,t2 =-9.68,P＜0.01).(3)The psychological stress,fatigue degree,confidence level of self-assessment health status had significantly positive correlation(r =0.517,0.488,0.259,0.352,P＜ 0.01).(4) The psychological stress,fatigue degree,confidence level of self-assessment health status entered the PSQI total score regression equation,moreover the predictive variation amount was 36.1％.Conclusion The psychological stress status and fatigue level in the militaries at plateau affect the sleep quality,and the confidence degree and health status assessment of officers and soldiers on exercise also produce the active influence on sleep.

Objective To analyze the main ingredients and investigate the anti-psoriasis effect in the standard decoction of Lithospermum. Methods The extraction rate was determined by extract determination method, and the content of total polysaccharide and total phenolic acid was evaluated by spectrophotometry. Moreover, the effects of different concentrations of the standard decoction of Lithospermum on skin lesions induced by imiquimod (IMQ) in psoriatic mice were observed. Results The extracting rate was 4.65 ％, the total polysaccharide content was 1.182 mg/ml and the total phenolic acid content was 3.506 mg/ml. The different concentrations of the standard decoction of Lithospermum could ameliorate the scales, erythema and psoriasis-like mice skin and reduce the thickness of epidermis. Conclusions The standard decoction of Lithospermum could improve the psoriasis-like lesions induced by imiquimod in mice and possess the anti-psoriasis effect.

To investigate the RNA interference (RNAi) effect induced by vector-derived small interfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online,we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene.Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different,depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80.

Objective To study the quality standard of Qingre-Chushi granules, including TLC identification of main components and ultra performance liquid chromatography determination of main components, so as to lay a foundation for further development of new drugs. Methods Seven constituents, including Gentianae Radix et Razoma, Scutellaria Radix, Plantaginis Herba, Rehmanniae Radix, Imperarae Rhizoma, Poria and Glycyrrhiza Radix et Rhizoma, were determined by TLC. The content of gentiopicroside and Baicalin in Gentiana officinalis and Scutellaria baicalensis was determined by UPLC on Acquity UPLC HSS T3C18 column (2.1 mm×100 mm, 1.7 μm) with acetonitrile-0.01％ formic acid water as mobile phase and gradient elution. Results TLC was highly specific and could effectively identify Gentian, Scutellaria baicalensis, Plantain, Rehmannia glutinosa, Rhizoma cocos, and licorice. The results of UPLC assay showed that there was a good linear relationship between gentiopicroside and Baicalin in the range of 0.014-0.028 μg and 0.015-0.449 μg, and the recovery of the three concentrations were all up to standard. Conclusions TLC can be used to identify the seven components of Qingre-Chushi granules, and UPLC can be used to determine the contents of three indicators, which can control the quality of Qingre-Chushi granules.

Objective:To observe the healing morphology, macular microstructure and visual function of idiopathic macular hole (IMH) after pars plana vitrectomy (PPV) combined with internal limiting membrane (ILM) flap.Methods:Retrospective case study. From 2016 to 2018, 39 eyes of 39 patients with IMH diagnosed in Tianjin Eye Hospital were included in the study. Among them, there were 4 eyes in 4 males and 35 eyes in 35 females, with an average age of 64.56±7.2 years. BCVA, OCT, OCT angiography (OCTA) and MAIA microperimetry examination were performed in all patients. BCVA examination was performed with the international standard visual acuity chart, which was converted to logMAR visual acuity when recording. All patients underwent PPV combined with ILM flap covering and air tamponade. According to the characteristics of OCT images postoperatively, the eyes were divided into U-shaped closed group, V-shaped closed group, irregular closed group and flat closed group, with 26, 5, 7 and 1 eyes respectively. There was a significant difference in the minimum hole diameter ( F=5.118, P=0.005) and macular hole classification ( F=3.608, P=0.024). The shallow capillary layer (SCP) blood flow density in the U-shaped closure group was significantly higher than that in the V-shaped closure group, the irregular closure group and the flat closure group (t=2.079, 2.368; P=0.047, 0.025). At 1, 3, 6 months after the operation, the same equipment and methods were used for relevant examination. The blood flow density of BCVA, SCP, perimeter of foveal avascular zone (PERIM) and mean sensitivity (MS) were compared before and after operation. Independent sample t-test was used for quantitative data comparison between different groups, and χ2 test was used for counting data comparison. Results:Six months after operation, the logMAR of the eyes in the U-shaped closure group was -0.75±0.29 higher than that before operation, and was better than that in the V-shaped closure group, the irregular closure group and the flat closure group ( t=-2.974, -2.518; P=0.006, 0.018). The integrity of external limiting membrane (ELM) and ellipsoid in U-shaped closed group was significantly higher than that in V-shaped closed group, irregular closed group and flat closed group ( χ2=15.229, 10.809; P=0.020, 0.013). The percentage of macular central fovea reflex mass in the U-shaped closed group was significantly lower than that in the V-shaped closed group, irregular closed group and flat closed group ( χ2=20.107, P=0.000). PERIM in U-shaped closure group was smaller than that in V-shaped closure group, irregular closure group and flat closure group, and the difference was statistically significant ( t=-3.391, -2.427; P=0.002, 0.022). The total MS of macular area 10° in the U-shaped closure group was significantly higher than that in the other V-shaped closure group, irregular closure group and flat closure group ( t=2.939, 2.811; P=0.001, 0.001). Conclusion:After IMH operation, the U-shaped closure showed better BCVA and macular light sensitivity, the proportion of ELM and ellipsoid to restore structural integrity are higher, PERIM is smaller, and there are fewer macular fovea strong reflex masses.

Objective:To analyze the distribution characteristics of special types of diabetes in China, in order to provide a theoretical basis for the diagnosis and treatment of special types of diabetes.Methods:Pubmed, CNKI, and WanFang Data were searched for the case reports and clinical studies of special types of diabetes in China from 2011 to 2021. After independent literature screening by 2 researchers according to the inclusion and exclusion criteria, diseases and the number of corresponding cases included were extracted for statistics. The etiological composition and disease characteristics of three subtypes of special type diabetes were analyzed.Results:A total of 613 articles(7 377 patients)were included and roughly divided into eight subtypes of special type diabetes according to etiological classification for disease composition analysis. The results by ratio in descending order were as follows: mono-genetic gene defects in islet β-cell function, pancreatogenic diabetes, diabetes induced by drugs or chemicals, endocrine disease, mono-genetic gene defects in insulin action, other genetic syndromes associated with diabetes, infection, and uncommon immune-mediated diabetes. The disease composition of the three subtypes of special types of diabetes that we focused on were mono-genetic gene defects in islet β-cell function(50.21%), pancreatogenic diabetes(35.65%), and mono-genetic gene defects in insulin action(1.56%). The composition analysis of the special types of diabetes in each subtype showed that neonatal diabetes mellitus(NDM, n=1 749, 23.71%)and maturity onset diabetes in young(MODY, n=1 554, 21.07%)accounted for the largest proportions. According to the composition analysis of each subtype of MODY patients, the top three subtypes were MODY2(50.89%), MODY3(16.03%), and MODYX(8.91%). In addition, taking MODY as an example, patients with de novo mutations(DNMs)and(or)new mutation sites were summarized and analyzed. The results revealed 31 MODY patients with DNMs(1.99%) and 339 MODY patients with new mutation sites(21.81%). Conclusions:According to the literature analysis, NDM and MODY represent the largest proportion of patients with special type diabetes in China. MODY2 patients make up the largest proportion of MODY patients. In addition, diabetic patients carrying DNMs and(or)new mutation sites should be taken seriously.

Congenital generalized lipodystrophy type 1 (CGL1) is an autosomal recessive genetic disease caused by mutations in AGPAT2 gene. The main clinical mainifestations include body subcutaneous fat loss, muscle hypertrophy, obvious subcutaneous veins, pseudoacromegaly, hirsutism, and acanthosis nigricans. What′s more, CGL1 is always accompanied by metabolic diseases. Therefore, it is easily misdiagnosed as metabolic syndrome, type 2 diabetes, polycystic ovary syndrome, acromegaly, or Cushing′s syndrome. Meanwhile, it is difficult to distinguish it from partial lipoatrophy syndrome. In this article, we present clinical and molecular characteristics of a patient with CGL1 and review mutations reported in literature to replenish current knowledge about this orphan disease.