Objective To observe the genetic predisposition of complement C5 gene polymorphisms in proliferative diabetic retinopathy (PDR) in Chongqing Han population.Methods 400 type 2 diabetes (T2D) patients (case group) and 600 age-and sex-matched healthy controls (control group) were enrolled in this study.There were 8 PDR patients in case group.All the subjects were Han ethnic people.The immune-related representative SNP locus of C5 gene including rs2269067,rs7040033,rs7027797 were screened by linkage disequilibrium analysis.Locus rs1017119 was selected by TagSNP and was around the above three loci.Subjects' peripheral venous blood was collected and DNA was extracted.Genotyping was examined by PCR-restriction fragment length polymorphism method.The level of C5 plasma protein was measured by enzyme-linked immunoabsorbent assay.Results The frequency of GG genotype of rs2269067 was significantly increased in PDR patients in cases group compared with controls (Pc=3.4 × 10-5,OR=1.87,95％CI=1.43-2.44;P=3.1 × 10-6).There was no differences in frequency of G,CC and CG genotype of rs2269067 between two groups (P=1.4 × 10-4,1.000,1.0 × 10-6).There were no differences in frequency of G,CC,CG,GG genotype of rs7040033,rs1017119,and rs7027797 between two groups (P＞0.05).The production of C5 plasma protein was significantly increased in case group as compare with control group (P=0.0004).An increased production of C5 plasma protein was observed in rs2269067 GG genotype cases compared to CG or CC cases (P=0.003,0.001).Conclusion C5 rs2269067 GG genotype may be associated with the PDR of T2D in Chongqing Han population.

Object To assess the genetic diversity among ten natural populations in Ophiopogon japonicus (L f ) Ker Gawl and O. bodinieri Levl (Liliaceae) in Sichuan Province Methods Forty random decamer primers were screened for Random Amplified Polymophic DNA (RAPD) fragments Results Based on cluster analysis of 515 DNA bands amplified by 11 primers, a DNA molecular dendrogram was established, and the relationship of the populations and seeds between O japonicus and O bodinieri was set up Conclusion 1 Distinct genetic differences and extensive genetic diversity are presented among the samples The genetic differences are related to morphological differences, but not to geographic regions; 2 Quite genetic differences are presented between O japonicus and O bodinieri, it suggests that they have farther relationships

Abstract: Objective　To analyze the incidence of pulmonary tuberculosis infection in the elderly(≥ 65 years old) in Xianning City from 2011 to 2021, and to provide scientific basis for formulating strategies and measures for tuberculosis prevention and treatment in the elderly. Methods　Descriptive epidemiological analysis was used to analyze the epidemic characteristics and influencing factors of elderly (this article studies elderly people aged 65 and above) pulmonary tuberculosis in Xianning City, Hubei Province from 2011 to 2021. Results　From 2011 to 2021, the average annual registration rate of elderly patients with pulmonary tuberculosis in Xianning City, Hubei Province was 211.52/100 000 (5 289/2 500 496), which showed a downward trend （χ2 trend=58.06, P<0.05), and significantly higher than the average registration rate of 96.04/100 000 patients with pulmonary tuberculosis (26 463/27 554 195)(χ2=2 896.69, P<0.05). The average proportion of the registered number of elderly patients with pulmonary tuberculosis in the whole population was 19.99% (5 289/26 463), which showed an upward trend (χ2trend=272.05, P<0.05). The average positive rate of etiology in elderly patients with pulmonary tuberculosis was 51.86% (2 743/5 289), which was significantly higher than that in the whole population 46.41% (12 282/26 463) (χ2=52.53, P<0.05). The average proportion of retreated patients in elderly patients with pulmonary tuberculosis was 8.92% (472/5 289), which was significantly higher than that of retreated patients in the whole population 6.33% (1 674/26 463) (χ2=47.23, P<0.05). The sex ratio of male to female in elderly patients with pulmonary tuberculosis was 2.92∶1. Among the six counties (districts) under the jurisdiction of Xianning City, Hubei Province the registration rate of elderly tuberculosis patients in Xian'an district was the highest 262.20/100 000 (1 267/483 220), and that in Tongcheng County was the lowest 123.83/100 000 (575/464 338), with significant differences (χ2=233.67, P<0.05). Conclusion　The overall incidence of tuberculosis in the elderly in Xianning City , Hubei Province showed a downward trend, but the proportion of the incidence of tuberculosis in the elderly in the whole population has increased year by year, suggesting that the elderly are the key population of tuberculosis prevention and control in Xianning City in the future, and comprehensive prevention and control measures should be taken to reduce the incidence of tuberculosis in the elderly population.

In this paper, the process performance indexes (PPIs) P(p) and P(pk) were introduced and applied to evaluate the process capability and quality consistence of Chinese medicine products. The historical quality analysis data of Qingkailing injection were collected and taken as the research object. The confidence intervals of P(p) and P(pk) were estimated based on the Bootstrap sampling methods. Results showed that the value and width of the confidence interval P(p) of were smaller that of P(pk) suggesting that the P(pk) index was more sensitive than P(p) in process capability analysis. Within the production period concerned, the P(pk) values estimated from different quality indexes of Qingkailing injection, such as baicalin, cholic acid, geniposide and total nitrogen, were 1.122, 2.055, 1.564 and 0.891, respectively. It could be found that the cholic acid had the highest process capability, followed by the geniposide and baicalin. The total nitrogen had the lowest process capability, indicating that it is necessary to reinforce the quality management of total nitrogen related manufacturing processes. The case studies demonstrate the effectiveness and feasibility of PPIs, which are convenient to be used in production practice of Chinese medicine.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; Injections ; Quality Control

Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.

Objective:To detect the effect of Celecoxib on the proliferation and apoptosis of human nasopha-ryngeal carcinoma cell line CNE-2. Method:The growth inhibition rate of CNE-2 by Celecoxib was evaluated with MTT method. Apoptosis related morphology changes were observed with transmission electron microscopy (TEM). The cell cycle and apoptosis were measured with flow cytometric method (FCM). Apoptotic index ( AI) was counted by the TDT-mediated dUTP-biotin nick end-labeling (TUNED assay. Result: The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner. Apoptosis with nuclear chromatin condensa-tion, cell shrinkage, periplast loss and the formation of apoptotic bodies was observed with TEM. Apoptotic rates of CNE-2 cells treated with 80 and 100 μmol/L celecoxib were (10. 47±0. 18)% and (20. 17±0. 55)% respective-ly, significantly higher than those of the control group (1. 57±0. 27)% with FCM. The percentage of G_0/G_1 phase cells increased, whereas the S and G_2/M phases cells decreased in a dose-dependent manner after the treatment. TUNEL assay showed that the apoptosis ratio( AI) of CNE-2 treated with Celecoxib was higher than control group (P<0. 01). Conclusion:Celecoxib can inhibit the growth of human nasopharyngeal carcinoma cell line CNE-2 and induce the cell apoptosis, which may be related to blocking the cell cycle progress of CNE-2 cells.

Background Oxygen-induced retinal neovascularization is the main pathological basis for many retinal vascular diseases.Research showed that light exposure at night can suppress retinal neovascularization in oxygen-induced retinopathy(OIR),but there were few reports discussing its effect on ROP.Objective This study aimed to observe the effect of light exposure at night on retinal neovascularization in an OIR mouse model.Methods Sixty-four newborn C57 BL/6J mice were randomly divided into four groups,with 16 mice for each group.OIR models were established by rearing the newborn C57BL/6J mice with their mothers in a(75±2)％ oxygen environment from postnatal day 7(P7)to Pl2,and then transferred to room air.In the OIR model group,the environmental illumination level was the same as the normal control group,and the model mice were exposed to 100 lx light at night in the OIR+ light exposure group.In the simple light exposure group,normal mice were reared in room air and were exposed to light at night from P12 to P17.All the mice were sacrificed on P17,and retinal flat mounts were prepared to assess the oxygen-induced changes of retinal vessels using the adenosine diphosphatase(ADPase)histochemical technique.The amount of proliferative neovascularization was quantified by counting the number of endotheliocyte nuclei in new vessels extending from the retinal inner limiting membrane into the vitreous in ocular cross-sections.The expression of the vascular endothelial growth factor(VEGF)protein was detected by immunohistochemistry.Real-time PCR analysis was performed to examine the expression of VEGF mRNA.The rearing and usage of the animals complied with the Statement of ARVO.Results Less free-vascular areas and new blood vessels were seen in the OIR+light exposure group compared with the OIR model group.On day 17 of the mouse life,the number of the endotheliocyte nuclei in new vessels extending from retinal inner limiting membrane were 0.97±0.83,1.00±0.72,38.57±5.01 and 16.92±3.39 in the normal group,simple light exposure group,OIR model group and OIR+light exposure group,respectively,showing significant differences among them(F =78.767,P =0.000).The number of nuclei in the OIR+light exposure group were less than that of the OIR model group(t=20.446,P＜0.01).Immunochemistry showed that the expression of VEGF in retina was weaker in the OIR+light exposure group than the OIR model group.The relative expression values of VEGF mRNA were 1.00±0.00,0.94±0.07,2.08±0.50 and 1.43±0.21 in the normal group,simple light exposure group,OIR model group and OIR+light exposure group,respectively,showing a significant difference (F=11.268,P =0.003),where the VEGF mRNA level in the OIR+light exposure group was lower than that of the OIR model group(t =20.163,P＜0.05).Conclusions Light exposure at night can weaken retinal neovascularization in OIR mice

OBJECTIVE To assess the efficancy of Yangxue Qingnao granules on simvastatin-induced vascular injury model in zebrafish. METHODS Since statins can inhibit vascular development in zebrafish,in this study,we developed a novel animal model using 1 to 3 day post-fertilization larval zebrafish by optimizing the doses and duration of simvastatin exposure.Five pro-angiogenic drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model. Zebrafish was treated with different concentration of Yangxue Qingnao granules( 62.5,125 and 250 μg·mL-1)for 2 d then tested for the area of subintestinal vein vessels. RESULTS Vascular regeneration promoting effect of five pro-angiogenic drugs (calycosin, astragaloside, chlorogenic acid, ferulic acid and Panax Notoginseng Saponins)were 8-48%,24-51%,35-58%,28-75% and 37-69%,respectively.In 62.5,125 and 250 μg·mL-1Yangxue Qingnao granules group,vascular regeneration promoting effect were 21% (P>0.05), 84%(P<0.01) and 53%(P<0.01). CONCLUSION Our results demonstrate that the zebrafish vascular injury model validated in this study could be used for in vivo angiogenesis studies and drug screening and for assessing pro-angiogenic drugs with different mechanisms.Yangxue Qingnao granules could promote the vascular regeneration in zebrafish.

Objective To estimate the value of computed tomography perfusion for clinical stage and approach the correlation of perfusion parameters and Cyfra21-1.Methods 63 patients with head and neck squamous carcinoma were confirmed by pathology and follow up underwent CT perfusion,which were divided into three groups by international clinical staging criteria(stageⅠ,stageⅡand stageⅢ-Ⅳ).BF,BV,MTT,TTP and Cyfra21-1 were recorded and compared with correlation in different clinical staging.Results There was no significant difference of Cyfra21-1 between stageⅠand stageⅡ(Z =1.439,P =0.1 62).There was significant differ-ence of Cyfra21-1 between stageⅠand stageⅢ-Ⅳ(Z =3.356,P =0.000),stageⅡand stageⅢ-Ⅳ(Z =4.959,P =0.000).There was significant difference of BF and BV between stageⅠand stageⅡ,stageⅠand stageⅢ-Ⅳ(P <0.05),of MTT and TTP between stageⅠand stageⅢ-Ⅳ,stageⅡand stageⅢ-Ⅳ(P <0.05).There was no significant difference of BF and BV between stageⅡand stageⅢ-Ⅳ(P >0.05),of MTT and TTP between stageⅠand stageⅡ(P >0.05).Cyfra21-1 and perfusion parameters in all groups have relationship(r=0.76,0.76,-0.82,-0.82,P <0.05).Conclusion The statistically significant of positive correlation be-tween Cyfra21-1 and perfusion parameters in head and neck squamous carcinoma suggests that CT perfusion could play a complemen-tary role in clinical assessment.

OBJECTIVETo develop an HPLC method for the determination of eleutheroside B in As1 (the extract fraction from Acanthopanax senticosus, which has a good effect of antifatigue).

METHODThe antifatigue effect of Asl was evaluated by mice burden swimming testing. One compound was isolated by silica gel, Sephadex LH-20 column chromatography and preparative high performance liquid chromatography. The structure was identified by physicochemical properties and spectral evidences. Eleutheroside B in Asl was determined by HPLC. Chromatographic conditions included Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase consisting of a mixture of acetonitrile-water-ethanoic acid (10: 90: 0.01). The UV detection wavelength was set at 344 nm.

RESULTAs1 showed an excellent antifatigue activity; One compound was isolated from As1 and its structure was identified as Eleutheroside B; The calibration cure was linear in the range of 0.104-20.8 microg (r = 0.9999), the average recovery was 97.68%, RSD 1.4% (n=6).

CONCLUSIONThis HPLC method is simple,accurate and reliable.

Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Fatigue ; drug therapy ; physiopathology ; Glucosides ; analysis ; Humans ; Male ; Mice ; Phenylpropionates ; analysis ; Swimming