Acute toxicity test of the Li-Dan-He-Ji granules / 中华危重病急救医学

OBJECTIVE@#To observe the acute toxic reaction of the Li-Dan-He-Ji granules, and to evaluate its safety.@*METHODS@#Sixty C57BL6/J mice were randomly divided into normal control group, vehicle group and drug treatment group, with 10 females and 10 males in each group. According to the Technical guidelines for the study of toxicity of single drug administration, the maximum administration dosage (MAD) was used to intragastric administration of Li-Dan-He-Ji granules 0.04 mL/g (42.8 g/kg), three times within 24 hours, with an interval of 6 hours. The vehicle group was fed with the same pure water. The normal control group received no treatment. The mice were observed continuously for 14 days, and the appearance characteristics, behavioral activities, body weight changes and the number of deaths in each group were recorded. At the 14 days, blood samples were collected from the eyeballs, and routine blood tests such as white blood cell count (WBC), lymphocyte count (LYM), neutrophil count (NEU), lymphocyte percentage (LYM%), neutrophil percentage (NEU%), red blood cell count (RBC), hemoglobin (Hb), and platelet count (PLT) were performed. And alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), creatinine (Cr) and other biochemical indicators. The mice were then sacrificed, and the histopathological changes of liver and kidney were observed by hematoxylin-eosin (HE) staining. The organ indexes of heart, liver, spleen, lung, kidney and thymus were calculated.@*RESULTS@#The median lethal dose (LD50) of Li-Dan-He-Ji granules were not obtained. During the MAD experiment, the animals in each group did not die, their behavioral activities were normal, and there was no significant change in liver and kidney histopathological examination. There were no significant differences in body weight, blood routine, biochemical indexes and organ index among all groups (all P > 0.05). The body weight (g) of normal control female and male group, vehicle female and male group and drug female and male group before administration were 18.96±1.14, 19.65±1.45, 19.33±1.30, 19.53±1.22, 19.28±1.69 and 19.48±1.28; 14 days after administration were 27.69±0.81, 28.19±2.22, 27.77±1.00, 27.88±1.85, 27.92±1.33 and 28.07±1.93, respectively.@*CONCLUSIONS@#The Li-Dan-He-Ji granules have low oral toxicity, combined with clinical observation, can be safely used in infants.

Analysis on Technical Problems and Countermeasures of Standard Formulation of Traditional Chinese Medicine Formula Granules / 中国实验方剂学杂志

After the completion of the pilot work of traditional Chinese medicine formula granules （TCMFGs）， the national and provincial medical products administrations have published and implemented about 440 varieties of TCMFGs standards. Based on the previous work， this paper analyzed technical problems encountered in the review and evaluation of Shandong TCMFGs standards， mainly involving the executive standards and distinguishing technologies of raw materials， the adding process of excipients in the procedure item， the rationality of quality control methods， the information content and reproducibility of characteristic chromatograms， the nomenclature and accessibility of reference materials， etc. The common problems such as the coverage of standards， specification differences， and the integrity of quality control items of current TCMFGs standards were discussed deeply. It is proposed to promote the upgrading of provincial standards to national standards， accelerate the research and development of reference materials， advocate the use of high-quality raw materials， explore the evaluation methods of high-quality products， and strengthen the quality supervision of the whole process. Suggestions of this paper is hoped to provide references for the formulation of national and provincial TCMFGs standards， promote the continuous improvement of TCMFGs standard system， and ensure the healthy and orderly development of the TCMFGs industry.

Neurological Diseases Responding Specifically to Traditional Chinese Medicine / 中国实验方剂学杂志

In recent years， the incidence of neurological diseases has been increasing year by year. To give full play to the advantages of traditional Chinese medicine （TCM） in the treatment of neurological disorders， identify the breakthrough point of integrating TCM with western medicine， and further standardize the clinical diagnosis and treatment of TCM， the China Association of Chinese Medicine organized neurologists in TCM and western medicine to carry out in-depth discussion on the neurological diseases responding specifically to TCM and integrated TCM and western medicine， such as stroke， headache， vertigo， multiple sclerosis， and epilepsy， aiming to formulate a well-recognized and integrated treatment protocol for TCM and western medicine and improve the efficacy of neurological disorders. Furthermore， the treatment suggestions of the corresponding diseases in TCM and western medicine were proposed to provide references for clinical practice and scientific research.

Sesquiterpenes and polyphenols with glucose-uptake stimulatory and antioxidant activities from the medicinal mushroom Sanghuangporus sanghuang / 中国天然药物

A chemical investigation on the fermentation products of Sanghuangporus sanghuang led to the isolation and identification of fourteen secondary metabolites (1-14) including eight sesquiterpenoids (1-8) and six polyphenols (9-14). Compounds 1-3 were sesquiterpenes with new structures which were elucidated based on NMR spectroscopy, high resolution mass spectrometry (HRMS) and electronic circular dichroism (ECD) data. All the isolates were tested for their stimulation effects on glucose uptake in insulin-resistant HepG2 cells, and cellular antioxidant activity. Compounds 9-12 were subjected to molecular docking experiment to primarily evaluate their anti-coronavirus (SARS-CoV-2) activity. As a result, compounds 9-12 were found to increase the glucose uptake of insulin-resistant HepG2 cells by 18.1%, 62.7%, 33.7% and 21.4% at the dose of 50 μmol·L

Research progress on preclinical efficacy evaluation of COVID-19 complicated with cytokine storm / 中国药理学通报

Studies have shown that COVID-19 patients infected with SARS-CoV-2 have severe pulmonary inflammation and cytokine storm, so the treatment of cytokine storm is an important part of rescuing critically ill patients with COVID-19. As an important cause of death, the preclinical study of cytokine storm is essential, and related experiments in vivo and in vitro are also the only way to develop new drugs for COVID-19 in the future. This paper reviews the in vitro and in vivo experimental methods of cytokine storm research articles at home and abroad in recent years, including the establishment of animal models, cell evaluation methods, pharmacodynamic evaluation indicators, etc., in order to provide reference and guidance for the experimental design methods of cytokine storm.

Psoralen and isopsoralen improve lipid metabolism disorder via inhibition of NF-κB activation in LO2 cells / 中国中药杂志

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using palmitic acid( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Efficacy of balloon Eustachian tuboplasty combined with grommet insertion in the treatment of chronic dilation Eustachian tube dysfunction / 临床耳鼻咽喉头颈外科杂志

OBJECTIVES@#To evaluate the efficacy of balloon Eustachian tuboplasty (BET) combined with grommet insertion in the treatment of chronic dilation Eustachian tube dysfunction (CDETD).@*METHODS@#A retrospective study was performed in 19 patients with CDETD who underwent BET at the Department of Otolaryngology-Head and Neck Surgery, Xinhua Hospital Affiliated with Shanghai Jiaotong University School of Medicine, from October, 2014 to September, 2016. The ages of these patients ranged from 10 to 67 years. All the patients underwent the preoperative assessment of oto-endoscope, tympanometry, pure tone audiometry, fiber nasopharyngeal endoscopy, Eustachian tube pressure measurement (TMM), CT and MRI. These patients had failed to respond to medicine, multiple tympanic membrane puncture and at least 2 times grommet insertion before our study. BET was performed in 5 patients (5 ears), and BET+grommet insertionwas performed in other 14 patients (23 ears). The changes of Eustachian tube function in these patients was assessed using the Eustachian tube score (ETS) and Eustachian tube dysfunction questionnaire-7 (ETDQ-7) preoperatively and 1, 3, 6, 9 and 12 months after surgery, respectively. In addition, subjective symptoms including the difficulty level of valsalva, aural fullness and earache were assessed by visual rating scale (VAS score) preoperatively and at 1, 6, and 12 months after surgery. The mean scores before surgery were compared with that at 1, 3, 6, 9 and 12 months. Postoperative adverse reactions and complications were recorded, such as earache, nosebleeding and so on.@*RESULTS@#Valsalva score and VAS score for aural fullness before surgery were 8.286±0.189 and 8.571±0.221, respectively. Valsalva score and VAS score for aural fullness were 3.714±0.317, 2.393±0.434, respectively, at one month after surgery, which were decreased significantly, as compared with the scores before surgery (<0.05). VAS score at 6 months and 12 months after surgery were statistically significant compared with those before surgery (<0.05). ETS score after surgery was significantly higher than that before surgery (<0.05). ETDQ-7 score after surgery was significantly lower than that before surgery (<0.05). The subjective satisfaction in these patients was 84.2%.@*CONCLUSIONS@#BET is simple and safe, with fewer complications, and effective for the treatment of CDETD combined with grommet insertion.

Research progress on influence of intestinal microflora on peripheral immune organs in mice / 中国免疫学杂志

The study found that the presence of intestinal microbiota is not only important for the metabolism of essential nutrients in the body, but also plays a key role in the development of the body′s immune system in recent years. Partial microbiota, through natural selection and co-evolution with the host, forms symbiotic relationships with host microbes that are inseparable from host physiology in mice. Symbiotic flora affects the formation of the body′s immune system by affecting innate and adaptive immunity and the development of various regulatory mechanisms. The destruction of the microbial ecosystem in the intestine can lead to the occurrence of many diseases,especially those related to the immune system. Peripheral immune organs always receive a number of immune cells colonized by antigen stimulation. So,the intestinal flora plays an important role in maintaining the function of immune cells. This article will investigates the effects of mouse-related intestinal flora on peripheral immune organ function.

The regulation effect of emodin on human embryonic hepatocyte L02 cell strain Farnesoid X receptor / 中华实用儿科临床杂志

Objective To investigate the regulation effect of emodin on human embryonic liver L02 cells strain farnesoid X receptor (FXR) pathways.Methods By using Guggulsterones,FXR genes were intervened with in L02 cells as model group,in three different concentrations of emodin (50.0 μ mol/L,25.0 μmol/L,12.5 μmol/L) of emodin in the model group cells,FXR,small heterodimer parter (SHP),UDP-glucuronosyltransferase 2B4 (UGT2 B4),bile salt export pump(BSEP) mRNA and protein expressions were detected by real-time fluorescent quantitative PCR and Western blot test.Results (1) The relative expressions of FXR mRNA and protein in the model group (0.240 ± 0.021,0.385 ±0.119) decreased significantly than those of control group (1.000 ± 0.088,1.000 ± 0.223),the differences were statistically significant (t =14.62,4.21,all P ＜ 0.01).Compared with the model group,the relative expressions of FXR mRNA in the high,the middle and the low-dose emodin groups (0.755 ±0.083,0.817 ±0.097,0.547 ± 0.080) were significantly higher (t =10.42,10.03,6.39,all P ＜ 0.01).The relative expressions of FXR protein in the medium-dose group (0.865 ± 0.203) increased significantly (t =3.53,P ＜ 0.01).The relative expressions of FXR mRNA in the high and the medium-dose groups (0.755 ± 0.083,0.817 ± 0.097) were higher than those in the low-dose group (0.547 ± 0.080),the differences were statistically significant (t =3.11,3.70,all P ＜ 0.01).(2)The relative expressions of SHP,UGT2B4,BSEP mRNA and protein in the model group (0.148 ±0.025,0.205 ± 0.039,0.184 ± 0.020;0.458 ± 0.130,0.255 ± 0.170,0.303 ± 0.100) were significantly lower than those in the control group (1.000 ±.0.099,1.000 ±0.104,1.000 ±0.125;1.000 ±0.129,1.000 ±0.157,1.000 ±0.162),the differences were statistically significant (t =14.50,12.44,11.19,5.13,5.57,6.33,all P ＜ 0.01).The relative expressions of SHP,UGT2B4 and BSEP mRNA in the high,the middle and the low-dose groups (0.610 ± 0.058,0.514 ± 0.041,0.707 ± 0.062;0.755 ± 0.108,0.800 ± 0.086,0.727 ± 0.076;0.470 ± 0.070,0.582 ± 0.050,0.500±0.108) were significantly lower than those in the model group (0.148 ± 0.025,0.205 ± 0.039,0.184 ± 0.020),the differences were statistically significant (t =12.75,9.38,13.94,9.46,10.90,11.96,7.53,10.31,5.00,all P ＜0.01).The relative expressions of SHP,UGT2B4 and BSEP mRNA in the high and the middle-dose emodin group (0.658 ±0.091,0.624 ±0.113,0.607 ±0.097;0.868 ±0.194,0.883 ±0.099,0.913 ±0.131) were significantly higher than those in the low-dose group (0.458 ±0.130,0.255 ±0.170,0.303 ±0.100),the differences were statistically significant (t =2.18,3.13,3.78,3.05,5.53,6.41,all P ＜ 0.01).The relative expression of SHP and BSEP protein in the low-dose group (0.645 ±0.135,0.572 ±0.076) increased,the differences were statistically significant (t =1.73,P ＜ 0.05,t =3.72,P ＜ 0.01).The relative expression of BSEP protein in the high and the medium-dose groups (0.607 ±0.097,0.913 ± 0.131) was significantly higher than those in the low-dose group (0.572 ± 0.076),the differences were statistically significant (t =1.99,3.90,all P ＜ 0.01).The relative expressions of SHP and UGT2B4 mRNA in the high and the low-dose group (0.610 ±0.058,0.470 ±0.070;0.514 ± 0.041,0.582 ± 0.050) were significantly lower than those in the medium-dose group (0.800 ± 0.086),the differences were statistically significant (t =3.75,6.47,3.83,3.42,all P ＜ 0.01).The expression levels of UGT2B4 and BSEP in the high and the low-dose groups (0.624 ± 0.113,0.644 ± 0.097;0.607 ± 0.097,0.572 ± 0.076) were significantly lower than those in the medium-dose group (0.883 ± 0.099,0.913 ± 0.131),the differences were statistically significant (t =4.27,2.98,6.30,3.90,all P ＜ 0.01).Conclusions Guggulsterones can inhibit FXR and downstream genes SHP,UGT2B4,BSEP expressions in L02,and emodin can enhance FXR gene expression,promote SHP,UGT2B4,BSEP gene expression,inhibit cholestasis pathway,protection of liver cell,which shows a dosage discreapancy.

Application of individualized nursing and double loop learning in early onset severe preeclampsia / 中国实用护理杂志

Objective Combining the concept of double loop learning in enterprise management with individualized nursing practice,the application value of this method in nursing care of patients with early onset severe preeclampsia(ESPE)was analyzed. Methods Patients with ESPE were selected from January 2014 to December 2016, which were divided into observation group and control group by random digits table method. The patients in observation group were given individualized nursing care with the idea of double loop learning,while the patients in control group received routine nursing care. The hospital admission, maternal pregnancy outcome and nursing satisfaction of the patients were compared between two groups. Results The observation group included 96 cases and the control group included 101 cases. There was no significant difference between two groups in age, gestational age, systolic blood pressure, diastolic blood pressure, and 24 hours urine protein(P>0.05).There were 87 cases of cesarean section in the observation group,while 94 cases in the control group, and there was no significant difference(P>0.05). The gestational weeks,the incidence of neonatal asphyxia,the nursing satisfaction was(35.5±2.7)weeks,5.2%(5/96), 90.6%(87/96)in the observation group and(34.1±2.0)weeks, 15.8%(16/101),72.3%(73/101)in the control group, the difference was statistically significant(t=4.149, χ2=5.843, 10.862, P<0.05 or 0.01). Conclusions In the ESPE nursing management, the individualized nursing and double loop learning can improve the child pregnancy outcomes and nursing satisfaction.

Study of expression and regulation of TLR2/4 in mycobacterium tuberculosis heat shock proteins 16. 3 effect on mouse bone marrow-derived macrophages / 中国免疫学杂志

Objective:To study the expression and regulation of TLR2/4 in mycobacterium tuberculosis heat shock proteins 16. 3 (mycobacterium tuberculosis heat shock proteins 16. 3,MTB Hsp16. 3) effect on mouse bone marrow-derived macrophages in vitro. Methods:Bone marrow cells were isolated from tibia and femurs of BALB/c mice and incubated with GM-CSF,then detected the expression of CD11b and F4/80 with flow cytometry and observed morphology. The M0 macrophages were stimulated with MTB Hsp16. 3 for 0 h,12 h,24 h,36 h,48 h and 72 h. Real-time PCR detected the expression of TLR2/4 in intracellular at different time point. Silencing macrophages cell surface TLR2/4 molecules by siRNA technology which stimulated with MTB Hsp16. 3 for 0 h,12 h,24 h,36 h,48 h and 72 h. Real-time PCR detected the expression of TLR2/4,Ym-1,Fizz1,IL-10,TNF-α,iNOS and TGF-βin intracellular at different time point. Results:Morphology analysis showed that MTB Hsp16. 3 stimulated macrophages were round cells stretching out pseudopodia,whereas MTB Hsp16. 3 stimulated silencing TLR2/4 macrophages had elongated fibroblastoid. Real time PCR detected the expression of TLR2/4 were upregulated after MTB Hsp16. 3 stimulated M0 macrophages. MTB Hsp16. 3 stimulated silencing TLR2/4 macrophages the expression of IL-6, TNF-α, iNOS were upregulated, whereas IL-10, TGF-β, Ym-1 and Fizz1 were downregulated. Conclusion:MTB Hsp16. 3 may stimulated M0 macrophages to M2 macrophages and suppress M1 macrophages through binding with TLR2/4 receptor,which may be involved the progresss of MTB evaded macrophage phagocytosis.

A study on the effect of regular aerobic exercise on artery elasticity among patients with essential hypertension / 预防医学

Objective To explore the effect of aerobic exercise training on artery elasticity among patients with essential hypertension.Methods A total of 155 cases of patients with essential hypertension (EH)were selected,and were divided into control group(76 cases)and aerobic exercise group(79 cases).Control group was follow routine drug therapy,and aerobic exercise group was follow aerobic exercise training besides the routine therapy for 12 weeks.CAVI,ABI values,limb blood pressure,TG,TC,LDL -C,HDL -C and U -MA were compared before and after the intervention.Results After the intervention,bilateral CAVI values of aerobic exercise group were 7.76 ±0.15,7.88 ±0.15,8.87 ±0.25 and 8.89 ± 0.22,and lower than those before the intervention(P <0.05).The decline range of bilateral CAVI value(the difference between the mean differences before and after the intervention)were 1.03 ±0.07 and 1.03 ±0.06,significantly wider than the control group of 0.33 ±1.97 and 0.15 ±0.08(P <0.05).After the intervention,limb systolic pressure,diastolic blood pressure and triglyceride,cholesterol decreased in the aerobic exercise group(P <0.05).The limb systolic blood pressure and left limb diastolic blood pressure decreased in the control group after intervention (P <0.05),but the decline range was smaller than the aerobic exercise group(P <0.05 ).Conclusion The aerobic exercise training could significantly improve arterial elasticity among patients with essential hypertension.

Protective effect of emodin pretreatment in young rats with intrahepatic cholestasis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of emodin in young rats with intrahepatic cholestasis.</p><p><b>METHODS</b>A total of 120 young Sprague-Dawley rats were randomly divided into control, model, and high-, medium-, and low-dose emodin groups, with 24 rats in each group. The rats in the control and model groups were given sodium carboxymethyl cellulose solution by gavage, while the other groups were given different doses of emodin solution by gavage. On the 5th day of experiment, alpha-naphthylisothiocyanate (ANIT, 50 mg/kg) was applied by gavage to establish the model of intrahepatic cholestasis in all groups except the control group. At 24, 48, and 72 hours after gavage, 8 rats in each group were sacrificed. Colorimetry was used to measure the serum levels of total bilirubin (TBIL), direct bilirubin (DBIL), total bile acid (TBA), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (GGT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in each group, and hematoxylin-eosin staining was applied to observe the morphological changes of the liver under a light microscope at different time points.</p><p><b>RESULTS</b>Compared with the control group, the model group had significantly increased serum levels of TBIL, DBIL, TBA, ALP, GGT, ALT, and AST at the 24-hour, 48-hour, and 72-hour time points (P<0.01). In the model group, the serum levels of TBIL, DBIL, TBA, ALT, and AST showed varying degrees of increase at 48 hours after establishment of model, compared with the values at 24 and 72 hours (P<0.05). At 24, 48, and 72 hours, the high-, medium-, and low-dose emodin groups had varying degrees of reductions in the serum levels of TBIL and TBA compared with the model group (P<0.05); the high- and low-dose emodin groups had significantly increased serum levels of TBA compared with the medium-dose emodin group (P<0.05). The model group had the most severe pathological changes at 48 hours. Compared with the model group, the high-, medium-, and low-dose emodin groups showed certain improvement in pathological changes of the liver at each time point, and the medium-dose emodin group had better improvement compared with the high- and low-dose emodin groups.</p><p><b>CONCLUSIONS</b>Emodin can effectively improve ANIT-induced intrahepatic cholestasis in young rats, and medium-dose emodin shows the best effect.</p>

Effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats

Objective: To observe effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats. Methods: A total of 75 SD rats were randomly divided into A, B, C, D, E groups with 15 in each group. Rats in group A served as the control group received just only but tissue separation without modeling operation, while model of unilateral ureteral obstruction (UUO) was established in B, C, D, E groups. Rats in A, B group were given saline lavage placebo treatment, while rats in C, D, E groups were given diammonium glycyrrhizinate and alprostadil injection. Five rats were sacrificed 1, 2, 3 weeks after modeling, serum creatinine level of femoral venous blood was determined. Transforming growth factor - β1 (TGF - β1) and concentration of connective tissue growth factor (CTGF) were also detected by using ELISA. Line renal interstitial tissue was taken after HE staining, renal interstitial TGF - β1 and CTGF expression were detected by using immunohistochemical method. Results: Serum creatinine levels of B, C, D, E group at different time points in were significantly higher than that of group A (. P0.05); while serum and kidney tissue TGF - β1, concentration of CREA, expression of rats in B, C, D, E groups showed a gradual increasing trend over time. TGF - β1 and CREF of Group B in serum and kidney tissues at each time point were significantly higher than that of the other groups (. P<0.05). TGF - β1 and CREF of Group E in serum and kidney tissues at each time point were significantly lower than that of B, C, D group at all time points in serum and kidney tissues (. P<0.05). Conclusions: Alprostadil combined with diammonium glycyrrhizinate can significantly lower the expression of TGF - β1 and CTGF in serum and tissues of SD rat with renal interstitial fibrosis, thus inhibit rat renal interstitial fibrosis process. It has synergy protective effect.

Effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats

OBJECTIVE@#To observe effect of alprostadil combined with Diammonium glycyrrhizinate on renal interstitial fibrosis in SD rats.@*METHODS@#A total of 75 SD rats were randomly divided into A, B, C, D, E groups with 15 in each group. Rats in group A served as the control group received just only but tissue separation without modeling operation, while model of unilateral ureteral obstruction (UUO) was established in B, C, D, E groups. Rats in A, B group were given saline lavage placebo treatment, while rats in C, D, E groups were given diammonium glycyrrhizinate and alprostadil injection. Five rats were sacrificed 1, 2, 3 weeks after modeling, serum creatinine level of femoral venous blood was determined. Transforming growth factor - β1 (TGF - β1) and concentration of connective tissue growth factor (CTGF) were also detected by using ELISA. Line renal interstitial tissue was taken after HE staining, renal interstitial TGF - β1 and CTGF expression were detected by using immunohistochemical method.@*RESULTS@#Serum creatinine levels of B, C, D, E group at different time points in were significantly higher than that of group A (P0.05); while serum and kidney tissue TGF - β1, concentration of CREA, expression of rats in B, C, D, E groups showed a gradual increasing trend over time. TGF - β1 and CREF of Group B in serum and kidney tissues at each time point were significantly higher than that of the other groups (P<0.05). TGF - β1 and CREF of Group E in serum and kidney tissues at each time point were significantly lower than that of B, C, D group at all time points in serum and kidney tissues (P<0.05).@*CONCLUSIONS@#Alprostadil combined with diammonium glycyrrhizinate can significantly lower the expression of TGF - β1 and CTGF in serum and tissues of SD rat with renal interstitial fibrosis, thus inhibit rat renal interstitial fibrosis process. It has synergy protective effect.

Construction and characterization of EGFP reporter gene labeled Sindbis virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct and characterize EGFP reporter gene labeled Sindbis virus (SINV).</p><p><b>METHODS</b>The reporter gene EGFP was inserted into the genome of infectious clone pBR-XJ160 by using multi-fusion long fragment PCR method. Then apply reverse genetic manipulation technique to rescue and obtain EGFP labeled SINV.</p><p><b>RESULTS</b>We successively obtained labeled SINV, which has good fluorescent expression characteristics and genetic stability.</p><p><b>CONCLUSION</b>The labeled virus can be seen in living cells and living body, and this serves as a good tool for cell and tissue tropism and biological function study of viruses. This study laid a foundation for further studying the cell tropism, biological functions and infection mechanism of SINV.</p>

Preparation and identification of a recombinant hepatitis C virus (HCV) based on sindbis virus vector / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct the recombinant virus-like particles containing HCV envelope glycoprotein E1E2 based on sindbis virus vector.</p><p><b>METHODS</b>The gene encoding HCV envelope glycoprotein E1E2 was cloned into sindbis virus vector to construct recombinant plasmids pBR-XJE1E2 and pVA-XJE1E2, and transfect them into BHK-21 cells to obtain recombinant virus-like particles. The expression of E1 and E2 protein were verified by Western Blot and indirect immunofluorescent assay (IFA).</p><p><b>RESULTS</b>The results of restriction enzyme digestion, PCR and sequencing analysis showed that the recombinant plasmids were constructed successfully. And the results of RT-PCR, Western blotting and IFA detection showed that the transfect cells could package HCV-like particles of expressing structural proteins E1E2.</p><p><b>CONCLUSION</b>The recombinant expression plasmids pBR-XJE1E2 and pVA-XJE1E2 based on sindbis virus vector could package HCV-like particles in eukaryotic cell, which provides a foundation for further study of its in vivo animal immune response.</p>

Phenotype and genotype analysis of three Chinese pedigrees with von Willebrand disease / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze phenotype and genotype of three Chinese pedigrees with von Willebrand disease (vWD), and explore the molecular mechanism.</p><p><b>METHODS</b>Bleeding time (BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation (RIPA), von Willebrand factor (vWF): ristocetin cofactor (RCof) (vWF:RCof), vWF antigen (vWF:Ag), vWF activity (vWF:A) test, vWF collagen binding assay (vWF:CB), vWF and Factor VIII (FVIII) binding assay (vWF:FVIII:B) and multimer analysis were used for phenotype diagnosis. Genomic DNA was extracted from the peripheral blood (PB). All the 52 exons and flanking sequences of the probands' vWF gene were amplified by PCR and analyzed by direct sequencing.</p><p><b>RESULTS</b>APTT were prolonged in all three probands, while BT were normal excepting for proband 3. Plasma RIPA, vWF:RCo, vWF:Ag, vWF:A and vWF:CB were decreased in different extents. In multimer analysis, proband 3 lost the large and intermediate molecular weight multimers, while proband 1 and 2 were normal. Gene analysis in the three probands revealed three heterozygous missense mutations of 144067 G→A (R2287Q) in exon 39, 110374G→A (R1374H) and 110770C→T (S1506L) in exon 28 and heterozygous polymorphism 110667G→A (D1472H) in exon 28, respectively.</p><p><b>CONCLUSION</b>The three heterozygous mutations (R2287Q, R1374H and S1506L) and an heterozygous polymorphism (D1472H) are genetic defects of the hereditary vWD of the three pedigrees respectively. R2287Q is a novel mutation reported for the first time in the literature.</p>

Molecular analysis of a patient with hemophilia A caused by FVIII His99Arg mutation / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the molecular mechanism of a Chinese hemophilia A patient in whom there was a discrepancy between the clinical bleeding symptoms and laboratory assay of FVIII activity (FVIII: C).</p><p><b>METHODS</b>FVIII: C was detected by chromogenic and one-stage methods, and FVIII: Ag by ELISA. The APTT corrected test was used to screen the FVIII inhibitor and PCR amplification to analyze all the exons and flanking sequences of F8 gene of the proband, PCR products were purified and sequenced directly. The corresponding gene sites of family members were detected according to the gene mutation sites. Two B domain deleted human FVIII mutant expression plasmids His99Arg and His99Ala (pRC/RS V - BDhFVIIIcDNA) were constructed and transfected into HEK293T transiently. FVIII: Ag and FVIII: C of the expression products were assayed.</p><p><b>RESULTS</b>The proband APTT was prolonged, FVIII: Ag was 120% but FVIII: C <1% and no FVIII inhibitor in plasma. The results of anticoagulation and fibrinolytic functions were normal. The cross reacting material positive (CRM+) hemophilia A was diagnosed. Gene analysis revealed a A28828G substitution in exon 3 resulted in a H (His) to R (Arg) missense mutation and the same heterozygous was identified in his mother. In vitro expression of FVIII: Ag and FVIII: C of His99Arg were 180.0% and 5.8% , respectively, while FVIII: Ag and FVIII: C of His99Ala were 45.0% and 20.0% of that of wild type, respectively. His99Arg and His99Ala were diagnosed as CRM+ and CRM- mutations, respectively.</p><p><b>CONCLUSION</b>Both the two F VIII mutations could express FVIII protein. However, CRM His99Arg mutant protein has little FVIII procoagulant activity and His99Ala has reduced FVIII function by routine methods.</p>

Molecular epidemiological analysis of KPC-2 and IMP-4 carbapenemase possessing Klebsiella pneumonia isolated from children / 中华检验医学杂志

Objective To investigate the distribution of acquired carbapenemases in carbapenemresistant strains of Klebsiella pneumoniae, and explore its role in epidemiology of nosocomial infection. Methods From November 2008 to March 2009, twenty clinical isolates of carbapenem-resistant Klebsiella pneumoniae were collected from children hospitalized in Wuhan children's hospital. MICs of antibiotics were tested by DNA of Klebsiella pneumoniae. Modified Hodge test was used to screen strains producing carbapenemases,combined imipenem(IPM)-EDTA , meropenem(MEM)- EDTA and ceftazidime(CAZ) - EDTA double-disk synergy test (DDST) were used to detect metallo-β-lactamase-producing. PCR amplification of the carbapenemase and integrase genes, and sequencing were performed. Plasmid conjugation transfer experiments and Southern hybridization were applied to study the mode of drug resistance transmission. Results Four types of Klebsiella pneumoniae were detected by PFGE, type A consisted of 5 strains, including 3 strains of type Al and 2 strains of type A2), type B (2 strains), type C (12strains) and type D (1 strain). Type A and C were the main drug resistant clones. Eight strains of Klebsiella pneumoniae carried both KPC-2 and IMP-4 genes, 10 strains carried IMP-4 gene, 2 strains carried KPC-2 gene. None of NDM-1 ,GIM, SPM, SIM, OXA-23, and VIM carbapenemase genes was detected in 20 isolates. All of 20 isolates carried lntl which were found to be located on bacterial chromosome by Southern blot. Conclusions KPC-2and IMP-4 genes are the major carbapenemase genes in Klebsiella pneumoniae isolated in Wuhan.Transmission of drug resistance is mainly through vertical transmission of type C resistant clone and horizontal transmission of Intl on bacteria chromosome.