Objective To isolate and identify embryonic stem (ES) cells of rhesus monkey from mature blastocysts cultured in vitro . Methods Rhesus monkey blastocysts were obtained after in vitro maturation of oocytes, fertilization and maturation of early embryos. After the blastocysts were hatched naturally from the zone pellucida, the inner cell mass (ICM) was dislodged from blastocysts using the sealed end of a finely drawn Pasteur pipette and co cultured with the feeder cells. The ICM, resembling embryonic stem cell colony, was isolated, cultured, and identified. Results A total of 92 normal GV oocytes were obtained from 4 FSH prime rhesus monkeys. Six blastocysts with high quality were selected from 22 oocytes after culture in HECM 10 medium. One of the rhesus monkey ES cell lines, i.e. RS5 cell line, was finally isolated from 3 of the inner cell mass isolated from the 6 blastocysts. The RS5 cells presented a high nucleus/cytoplasm ratio, prominent nucleoli, and flatter colonies with individual and distinct cells. After successive passages for 5 months passage, the RS5 cells remained a normal karyotype, i.e. 42 of chromosomes and alkaline phosphatase (AP) positive, which meant the RS5 ES cell colony remained undifferentiated. After high density culture for a longer time, the ES cell could differentiate into multi types of cells. Conclusion The RS5 cell line has the ability to self renew and potential to differentiate. Thus, RS5 cell line belongs to embryonic stem cells.

Humans ; Infant, Newborn ; Meconium ; Peritonitis ; etiology

Objective To evaluate the clinical efficacy of thalidomide plus mometasone furoate cream under occlusion and ultraviolet irradiation for the treatment of prurigo nodularis.Methods A non-randomized,parallel,controlled study was carried out.Eighty patients with prurigo nodularis were divided into 3 groups,i.e.,control group(no irradiation),ultraviolet A1(UVA1)group,and ultraviolet B(UVB)group.All the patients were treated with oral thalidomide and topical mometasone furoate cream under occlusion.Additionally,the patients in UVA1 group and UVB group received UVA1 and NB-UVB irradiation,respectively,thrice a week for no less than 8 weeks.Patients were evaluated at the baseline,and on day 30 after the beginning of treatment.Clinical outcome parameters included disease severity score and visual analogue scales for pruritus.Peripheral blood eosinophils were counted during each visit.Rank sum test was performed to compare the clinical efficacy between the 3 groups,and the relationship between peripheral blood eosinophile count and visual analogue scales for pruritus was analyzed.Results After 30 days of treatment,skin lesions were markedly improved in 5 (21.74％),13(43.33％)and 9(37.5％)patients,and improved in 7(30.43％),12(40％)and 7(29.17％)patients,in the control group,UVA1 group and UVB group respectively;a marked response in pruritus was noted in 7(30.43％),18(60.00％)and 14(58.33％)patients respectively in the control group,UVA1 group and UVB group.The efficacy on skin lesions and pruritus was significantly stronger in the UVA1 group and UVB group than in the control group(skin lesions:Z =8.21,5.22,both P ＜ 0.01;pruritus:Z =4.50,4.50,both P ＜ 0.01),but similar between the UVA1 group and UVB group(skin lesions:Z =0.50,P ＞ 0.05;pruritus:Z =0.35,P ＞ 0.05).Peripheral blood eosinophil count was positively correlated with the visual analogue scale for pruritus in the patients(r =0.53,P ＜ 0.01).Conclusions Thalidomide combined with mometasone furoate cream under occlusion and ultraviolet irradiation shows notable efficacy for the treatment of prurigo nodularis,and the combination with UVA1 or NB-UVB irradiation enhances the efficacy of thalidomide and mometasone furoate cream under occlusion.

Objective To develop a better probe preparation method of gene chips for the further gene chips fabrication and its application in the gene expression. Method Using human leukemia K562 cells as material,applying a new kind of technology for gene differential display restriction display(RD PCR),we isolated a lot of cDNA fragments as probes of gene chips. Results This method has overcome the shortcoming of more pseudopositives in DD PCR,and was easy to be manipulated.Compared with the chip fabricated with full cDNA probes,the probe length is comparatively homogeneous,and the hybridization dynamics is apt to be controlled.Conclusion\ RD PCR technology could provide a new strategy for the probe preparation of gene chips.\;[

Objective To study the chemical constituents of Helicia nilagirica. Methods The ethanol extract was seperated by petroleum ether, dichloromethane, and n-butanol in sequence, then isolated by silica gel column chromatography. The structures were identified and elucidated by physicochemical pro-(perties) and spectral analysis. Results Four compounds were isolated from the petroleum ether extract and identified as n-hexadecane acid (Ⅰ), ?-sitosterol (Ⅱ), ?-sitosterol-3-O-?-D-glucoside-6′-acetate (Ⅲ), and daucosterol (Ⅳ). Conclusion All the compounds are isolated from the plant for the first time. Compounds Ⅰ, Ⅲ, Ⅳ are isolated from the plants of Helicia Lour. for the first time, and compound Ⅲ is a new natural product.

OBJECTIVETo compare the efficacy difference in treatment of myofasical pain syndrome between sparrow-pecking moxibustion and acupuncture at trigger points so as to provide the reference of the effective therapeutic method for myofascial pain syndrome.

METHODSNinety patients were randomized into a sparrow-pecking moxibustion group and an acupuncture group, 45 cases in each one. The trigger points were selected in pain areas in the two groups. In the sparrow-pecking moxibustion group, the sparrow-pecking moxibustion was applied, 30 min in each time. In the acupuncture group, the filiform needles were inserted obliquely at 45 degrees and retained for 40 min in each treatment. The treatment was given once a day and 10 treatments made one session in the two groups. The short-form McGill pain questionnaire was used as the observation index, and the changes in pain rating index (PRI), present pain intensity (PPI) and visual analogue scale (VAS) before and after treatment were used for efficacy assessment.

RESULTSThe results of PRI, PPI and VAS after treatment were reduced apparently as compared with those before treatment in the sparrow-pecking moxibustion group and the acupuncture group (all P<0.001). The differences in PRI, PPI and VAS after treatment were not significant in comparison of the two groups (both P>0.05). The curative and remarkably effective rate was 80.0% (36/45) in the sparrow-pecking moxibustion group, which was better than 40.0% (18/45, P<0.001) in the acupuncture group.

CONCLUSIONSparrow-pecking moxibustion at trigger points achieves the superior efficacy on myofascial pain syndrome as compared with acupuncture at trigger points. This therapy is simpler in operation additionally.

Acupuncture Points ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Myofascial Pain Syndromes ; physiopathology ; therapy ; Treatment Outcome ; Trigger Points ; physiopathology ; Young Adult

AIM:To investigate the effect of pretreatment of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) on the proliferation and the differentiation of mesenchymal stem cells(MSCs) into cardiomyogenic cells.METHODS:The MSCs,isolated primarily from bone marrow,and purified by passage culture,were obtained from the adult rats of four groups:the rats were pretreated by 5 daily injections of SCF;the rats were pretreated with G-CSF;the rats were pretreated with SCF and G-CSF;the rats were treated without any intervention.The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co-culture.The neonatal rat cardiomyocytes cultured for 3 days were co-cultured with DAPI-MSCs.The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co-culture days was analyzed.The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were recorded respectively with digital microscope camera system and immunofluorescence technique.The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated.RESULTS:The percentage of MSCs in G0/G1 phase in SCF/G-CSF group was significantly lower than that in SCF group,G-CSF group and the control group.The percentage of MHC protein-positive MSCs in SCF/G-CSF group was markedly higher than that in SCF group,G-CSF group and the control group,and that in SCF group and G-CSF group was significantly higher than control group.The percentage of TnT protein-positive MSCs in SCF/G-CSF group,SCF group and G-CSF group was significantly higher than that in control group.CONCLUSION:SCF and G-CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes.The combination of using SCF and G-CSF is more effective than using only SCF or G-CSF.

Objective: To evaluate the diagnostic value of ¹⁸F-fluorodeoxyglucose (FDG) PET/CT imaging in non-traumatic unilateral vocal cord paralysis (UVCP) and compare the radioactive uptake in different lesions. Methods: Clinical data of 62 patients (49 males, 13 females; age: (61.7±12.8) years) with non-traumatic UVCP (43 cases of left vocal cord paralysis and 19 cases of right) admitted to Ji′an Hospital from January 2016 to December 2018 were analyzed retrospectively. Pathological results, imaging or follow-up results were considered as the standard of final diagnosis. The diagnostic efficacy of PET/CT imaging for the primary cause was analyzed. The maximum standardized uptake values (SUV_max) of vocal cord in patients with different etiology were compared by independent-sample t test. Results: According to the final diagnosis, the primary causes of UVCP were as follows: malignant tumors (n=44), inflammation (n=16), glomus jugulare tumor (n=1) and idiopathic UVCP (n=1). The diagnostic accuracy of PET/CT imaging for the primary cause was 90.32%(56/62): 44 cases were correctly diagnosed as malignant tumors, while 11 cases as inflammation, and 1 case as glomus jugulare tumor. Among 62 patients, 29 patients had increased SUV_max in the affected side (direct invasion group; further divided into tumor group (n=12) and non-tumor group (n=17)), and other 33 patients had increased SUV_max in the healthy side (indirect invasion group). SUV_max of the affected vocal cord in direct invasion group was higher than that in the healthy side (9.97±5.21 vs 2.43±0.62; t=8.14, P<0.01). The differences between affected side and healthy side in the tumor group (14.92±3.91 vs 2.84±0.54) and the non-tumor group (6.48±2.48 vs 2.14±0.50) were statistically significant (t values: 9.94, 7.93, both P<0.01). In the indirect invasion group, SUV_max in the affected side of vocal cord was significantly lower than the healthy side (1.89±0.35 vs 6.97±2.63; t=11.44, P<0.01). There were significant differences in radioactive uptake between affected side of direct invasion group and healthy side of indirect invasion group (t=2.86, P<0.01), affected sides of tumor group and non-tumor group (t=7.46, P<0.01), affected side of tumor group and healthy side of indirect invasion group (t=6.07, P<0.01). But the radioactive uptake difference between affected side of non-tumor group and healthy side of indirect invasion group was not statistically significant (t=0.51, P>0.05). Conclusions ¹⁸F-FDG PET/CT imaging has high diagnostic value in pathogenic diagnosis of non-traumatic UVCP. The different radioactive uptake of vocal cords in the affected side and the healthy side provides more accurate etiological information for clinical analysis.

Dietary Proteins ; administration & dosage ; Energy Intake ; Female ; Hospitalization ; Humans ; Infant, Newborn ; Infant, Very Low Birth Weight ; Logistic Models ; Male ; Parenteral Nutrition ; Weight Gain

Objective To observe microvascular architecture and free radical metabolism in hippocampus after focal cerebral ischemia/reperfusion and to explore the effect of NBP (3-n-butylphthalide). Methods Fifty-four SD rats were ran?domly divided into NBP pretreatment group, ischemia/reperfusion group and sham operation group (n=18 in each group). The model of middle cerebral artery occlusion(MCAO)was established by suture method. The neurological scores were counted and the volume of infarction was measured;TA-Fe method was applied to observe the microvascular architecture of hippo?campus, Mivnt image analysis system was used to analyze the microvessel density(MVD)and the microvessel area density (MVA)of hippocampus quantitatively;The activity of SOD and content of MDA were measured by colorimetric method. Re?sults Compared to the ischemia reperfusion(IR)group, the neurological scores and the volume of infarction were decreased sharply in NBP group. What′s more, the activity of SOD, MVD and MVA were all enhanced but the content of MDA and the count of closed microvessels were both reduced(P < 0.01). Conclusion NBP can improve microvascular architecture of hippocampus and reduce the free radical injury. There is a protective effect on hippocampus of rats who suffered focal cere?bral ischemia reperfusion.