Single cell RNA sequencing (scRNA-seq) is an advanced technology to study the transcriptome information at the single cell level. The application of this technology can attribute to analyze the heterogeneous map of cells in the process of disease development, and precisely identify the specific cell subsets that are responsive to pharmacological therapy. Currently, scRNA-seq technology has been widely applied in the field of drug research, including studies on therapeutic targets, drug-induced adverse reactions, drug resistance and vaccine. This work reviews the application of scRNA-seq technology in drug discovery, which offers a scientific basis for personalized and accurate medication therapy.

BACKGROUND: Bone marrow stem cells can be induced to differentiate into vascular endothelial cells and facilitate establishment of compensatory circulation, so as so improve blood supply of ischemic tissues. OBJECTIVE: To observe the clinical efficacy of autologous peripheral blood stem cells (PBSCs) transplantation in the treatment of diabetic vascular disorder in the lower extremities and diabetic feet. DESIGN: Wuhan Central Hospital, Wuban, Hubei Province, China. PARTICIPANTS: A total of 89 patients with type 2 diabetic vascular disorder in the lower extremity were enrolled from Department of Endocrinology in Wuhan Central Hospital from July 2005 to may 2007. All cases matched the criteria of diabetes by WHO in 1999, and complicated with diabetic peripheral angiopathy and diabetic feet. They included 61 males and 28 females, aged 45-90 years, with an average age of 67. Lesion was mainly located in lower extremities, 54 patients with foot ulcers while 8 patients with gangrene. All the patients signed the informed consents, and the experiment was approved by the hospital ethics committee. Recombinant human granulocyte colony stimulating factor was purchased from China Kunpeng Bio-Pharmaceutical Co., Ltd.METHODS: Stem cell mobilization: 89 patients all received recombinant human granulocyte colony stimulating factor, 500-600 μg per day by hypodermic injection for 5 days to mobilize stem cells. On the fifth day. PBSCs were collected with a total amount of 83-103mL. The number of mononuclear cells was (1.86-3.39)×1011L-1,while the proportion of CD34 positive cells was 0.55%-1.36%. PBSCs transplantation: After patients were treated with intravenous anesthesia, PBSCs were injected into the ischemic lower extremity and foot intramuscularly at 3cm×3cm distance. The clinical and laboratory findings were monitored from the first day to the sixth month.MAIN OUTCOME MEASURES: The patients were evaluated before transplantation, and in the first, third and sixth months after transplantation, including affected limb pain, cool feeling, intermittent claudication, ankle brachial index (ABI), limb skin temperature and deep feeling. Color Doppler. CT and digital subtraction angiography were applied to detect lower extremities morphous.RESULTS: All 89 included patients were involved in the result analysis. Scores of clinical symptom: The affected limb pain, cool feeling, and intermittent claudication were all improved significantly after PBSCs transplantation (P<0.05),and the improvement was more evident with time extension. ABI: After PBSCs transplantation, ABI in patients increased significantly by different degrees with the time extension(P0.05). Skin temperature and deep feeling: Limb skin temperature increased significantly after transplantation (P<0.05), and deep feeling seemed to be improved, but the change was not significant(P0.05). Imaging determination: There was no obvious change after transplantation checking of lower extremity by color Doppler. But CT and digital subtraction angiography results showed lower extremities in 23 patients had new collateral vessels formation with different degrees.CONCLUSION: Autologous PBSCs transplantation can increase blood flow of lower extremities in patients with diabetic vascular disorder in the lower extremities and diabetic feet, and promote the angiogenesis in lower extremity of partial patients.

Objective To supervise military training by application of clinical classification of fatigue fracture so as to decrease the occurrence and development of fatigue fracture. Methods From Feb.1999 to Feb.2002, the knowledge of clinical classification of fatigue fracture was applied in the military training in which it was essential to prevent and treat fatigue fracture. Results From 1999 to 2002, the incidence of fatigue fracture decreased year by year. Types Ⅱand Ⅲof fatigue fracture decreased significantly. Conclusion Clinical classification plays an important role in decreasing the occurrence and development of fatigue fracture.

Objective To investigate the usefulness of acoustic radiation force impulse (ARFI) elastography for noninvasive evaluation of liver fibrosis in rats.Methods A total of 70 male Wistar rats were included in the group for dimethylnitrosamine (DMN)-induced liver injury,and 10 saline-injected rats were used as normal control.Hepatic injury was induced by a single intraperitoneal injection of DMN at a dose of 50 mg/kg of body weight.Several rats in the group with DNM injected and the normal control group were randomly selected and sacrificed at each of the following post-injection time:day 5,7,10,14,21,24,and 28.And their livers were taken for pathology analysis.All the rats underwent ARFI elastography before sacrificed in order to acquire a shear wave velocity (Vs) to represent liver stiffness.Correlation between Vs and the histological finding was analysed.ResultsAmong 58 successfully modeled rats,9,13,14 and 12 rats were found to be with S1,S2,S3 and S4 of liver fibrosis pathologically,respectively.And 10 rats were found to be with severe inflammatory activity without any fibrosis.Values of Vs increased with the stage of liver fibrosis ( P ＜0.05).There was a significant correlation between Vs and stage of liver fibrosis ( r =0.947,P =0.000).The areas under ROC curve for the diagnosis of fibrosis S≥S1,S≥S2,S≥S3 and S=S4 were 0.983,0.995,0.999 and 0.964,respectively;for the cutoff values of Vs were 1.59 m/s,2.13 m/s,2.33 m/s and 2.51 m/s,respectively,the sensitivity was 95.8％,92.3％,100％ and 84.6％,and specificity was 100％,100％,96.9％ and 95.6％,respectively.The values of Vs in the group with severe inflammatory activity were significantly higher than those in the control group ( P =0.000).ConclusionsARFI has a relatively high value in the evaluation of liver fibrfosis in rats,while severe inflammatory activity may affect its accuracy.

A strain was separeted from the Yueqing bay using pine pollen baiting.The vegetative thallus of the separated strain is oval and unincleate.It possesses a cell wall composed of many compact layers of closely pressed scales, which can be resolved where the cell wall is disrupted.The radiating branched extensions of the thallus, the ectoplasmic net, emerges from the sagenogenetosome.Asexual reproduction is by conversion of the vegetative thallus to many biflagellate zoospores, during which tetrads of cells are formed.It was identified with Schizochytrium sp.based on the features mentioned above.

Aim To establish a model of the vascular endothelial cell (VEC) injury in SDrats.Methods SD rats were randomly divided into the control and the modelgroups. The model rats were injected with adrenaline diluted to 2. 5 times 0. 05 mg?100 g-1 (tid) for 5 d continously. From the 4th d, they were irritated for 5 min in the0℃ cold-water in the middle between adrenaline injections.The control rats weregiven 0. 9% NS as above. At 6th d, blood samples were taken from carotid arteries ofthe rats and the CEC counts, t - PA、PAI activities, 6-keto-PGF1? concentrations andthe platelet aggregation rate(max) were detected respectively. Results In the modelgroup, as compared with those in the control group, t - PA activity and 6-keto-PGF1?concentration decreased significantly(P

Objective To establish normal range of components of cord blood cells in healthy fetuses from 19 to 37 weeks' gestation and to provide proof for diagnosis of hematological disorders in prenatal fetuses and premature infants.Methods Twelve hematological parameters were determined in 182 fetuses using Coulter GENS system 2 full automated blood cell counter,and the blood cells were classified by microscope.Results The number of white blood cell(WBC) was increased gradually from 3.58?10~9/L to 5.76?10~9/L with the gestational weeks increasing from 19 to 37 weeks.The differential counts indicated that the lymphocytes represented the main population.The number of lymphocytes and normoblast was decreased gradually and that of neutrophils was increased gradually.The numbers of monocytes,eosinophils and basophils remained stable as the increasing of gestational weeks.The red blood cell(RBC),hemoglobin(HGB) and hematocrit(HCT) were increased gradually but mean corpuscular volume(MCV) was decreased.The differences between mean corpuscular hemoglobin(MCH),platelet volume distribution width(PDW),mean platelet volume(MPV),plateletcrit(PCT) and mean corpuscular hemoglobin concentration(MCHC) were not significant among different fetuses' ages.Conclusion The components of cord blood cells in healthy fetuses are dynamic and the establishment normal range of components of cord blood cells in healthy fetuses is helpful to diagnose the disorders in prenatal fetuses and premature infants.

Objective To evaluate the clinical values of acoustic radiation force impulse(ARFI) elastography for liver fibrosis on hepatopath patients.Methods ARFI elastography was prospectively performed on 99 patients prepared to undergo hepatic surgery to get a shear wave velocity(Vs) which was the representative of liver stiffness.The fibrosis stages,inflammation grades and steatosis grades were evaluated histologically after surgery.Values of Vs were compared with the histological results.Results Values of Vs with fibrosis stage 0-4 was (1.14 ± 0.22) m/s,(1.30 ± 0.22) m/s,(1.36 ± 0.38) m/s,(1.47 ± 0.37)m/s and (1.87 ± 0.08) m/s,respectively.A significant difference was observed among them (P ＜0.001).There was a certain correlation between Vs and fibrosis stage(r =0.541,P ＜0.001).Vs was a significant predictor of stage ≥3 fibrosis with an area under the ROC curve of 0.812,sensitivity 73.2％ and specificity 81.4％ when 1.40 m/s was the cutoff value (P ＜ 0.001).Values of Vs with inflammation grade 0-3 was (1.17 ± 0.16)m/s,(1.43 ± 0.36) m/s,(1.59 ± 0.53) m/s and (1.89 ± 0.59) m/s,respectively,which were significantly different (P ＜ 0.001).A certain correlation was observed between Vs and inflammation grade(r =0.383,P ＜0.001).Values of Vs with steatosis grade 0-4 was (1.61 ±0.42) m/s,(1.47±0.58) m/s,(1.56 ± 0.71)m/s,1.13 m/s and (0.94 ± 0.95) m/s.Obvious difference didn't exist between Vs and steatosis grade (P =0.286).Obvious correlation wasn't observed between them,either (r =-0.177,P =0.080).Conclusions ARFI elastography has a certain value for the evaluation of liver fibrosis,while inflammation grade may affect its performance.

Objective To explore the diagnostic value of virtual touch tissue quantification (VTQ) for the assessment of liver fibrosis in patients with chronic hepatitis B.Methods VTQ was prospectively performed in 181 patients with chronic hepatitis B and a shear wave velocity (Vs) indirectly reflecting liver stiffness was obtained for each patient.With the gold standards of fibrotic stages and inflammation grades,the relationship between Vs,fibrotic stages and inflammation grades was evaluated.Results A relatively high correlation existed between Vs and liver fibrotic stages (r ＝ 0.812,P ＜ 0.001).Significant differences existed among groups with different fibrotic stages except for groups SO vs.S1 and S1 vs.S2 (P ＜ 0.05).The areas under the ROC curve were 0.971,0.96l,0.969 and 0.908 for Vs predicting S≥S1,S≥S2,S≥S3 and S =S4 fibrosis respectively.Moderate correlation existed between Vs and inflammation grades (r =0.476,P =0.000).Conclusion Capable of staging liver fibrosis and preliminarily grading inflammation,VTQ is helpful for the early diagnosis of liver fibrosis and assessment of therapeutic follow-up and prognosis in patients with chronic hepatitis B.

OBJECTIVETo investigate the effects of soluble M-CSF receptor (sMR) on proliferation and differentiation of hematopoietic precursors derived from umbilical cord blood in mesenchymal stem cell (MSC) microenvironment.

METHODIn group of cytokine (CK) + sMR, MSCs were used as feeder cells, mononuclear cells (MNCs) from cord blood were expanded in MSC microenvironment in presence of SCF, Flt3L, TPO, IL-6 and sMR. In CK control group, no sMR was added. MNC counting and colony forming cell (CFC) culture were performed at week 1, 2, 3 and 4.

RESULTS1) The number of MNCs increased rapidly in both group CK and group CK + sMR (108.47 -fold and 120.67 -fold, respectively, P > 0.05). 2) CFC increased, peaked at week 3(38.1 x 10(3)) and declined rapidly at week 4(18.1 x 10(3)) in group CK, but still increased in group CK + sMR at week 4 (84 x 10(3)), the total number of CFC was higher in group CK + sMR than in group CK at week 3 and week 4 (P <0.01). 3) The erythroid CFC peaked at week 1 (5891.2 and 5635.6 for groups CK and CK + sMR, respectively), then dropped rapidly and to zero at week 3, in both group CK and group CK + sMR (P > 0. 05). 4) Myeloid CFC expanded continuously and peaked at week 3 (31.5 x 10(3)), then declined at week 4 (18.3 x 10(3)) in group CK; but still increased at week 4(80.8 x 10(3)) in group CK + sMR, being higher than that in group CK at week 3 and week 4 (P <0.01).

CONCLUSIONsMR can inhibit the differentiation of cord blood hematopoietic precursors expanded in MSC microenvironment, but the inhibition exerts only on myelomonocytic but not on erythroid precursors.

Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Mesenchymal Stromal Cells ; Receptor, Macrophage Colony-Stimulating Factor ; chemistry