Objective To establish the crystal violet plaque assay for detection of virus titer of recombined Tiantan vaccinia AIDS vaccine,and provide more stable method of virus titration for rTV AIDS Vaccine.Methods Optimized the concentration of Vero cells,the time and temperature of virus adsorption,and the time of determination for CPE,then established the crystal violet plaque assay for virus titer of rTV.Counting and analysis the plaques by BioSpot Reader,then analyzed the relativity of plaques counted with BioSpot Reader and manual; Several lots rTV AIDS Vaccine and Tiantan vaccinia were titrated by the method of plaque formation-hemadsorption assay,neutral red and crystal violet plaque assay,then analyzed the relativity of the results of three methods ; meanwhile,the virus titer of samples were determine repeatedly by the crystal violet plaque assay,then calculated the coefficient of variation( CV),and verified the precision of the method; SPSS17.0 was used in statistical analysis of the experimental results.Results When the concentration of Vero cells was 5.0×105-9.O×105 cells/ml,virus been adsorbesd 2 h at 37℃,then cultivated 72 h after adding the culture medium containing methyl cellulose.Plaques counted by BioSpot Reader was highly related with counted by manual (r =0.985),so BioSpot Reader counting can objectively reflect the virus plaques with various size,and reduce the error by manual counting; compared the virus titration for different lots of rTV AIDS vaccine and Tiantan vaccinia with three methods,the crystal violet plaque assay was highly related with plaque formation-hemadsorption assay (r =0.997,P＜0.01 ) and neutral red plaque assay(r=0.980,P＜0.01 ).Conclusion Crystal violet plaque assay was established for virus titration of rTV AIDS Vaccine.

Objective To study the genetic diversity of Fritillaria thunbergii,a traditional Chinese herb in Zhejiang Province in China.Methods The genetic diversity of six representational populations of F.thunbergii including 32 individuals was investigated by amplified fragment length polymorphism(AFLP) maker technique.Results The genetic diversity was revealed as follow: the Nei′s genetic diversity index(He) 0.169 0?0.175 7,Shannon′s information index(I) 0.269 8?0.245 3,percentage of polymorphic loci(PPB) was 76.85% at the species level;Ht 0.169 0?0.030 9,and Hs 0.150 8?0.024 0,I 0.233 3?0.261 9, PPB was 50.38% at population level.The genetic differentiation index(Gst) was 0.107 6,Nm 4.147 0.The result of dendrogram of six populations indicated that Dongyang and Yongkang populations shared the minimum genetic distance(0.015 0),they were classified into a group,and Xiangshan and Jinyun populations shared the maximum genetic distance(0.032 4).Conclusion The genetic diversity of F.thunbergii cultivated in Zhejiang Province is very rich,which could ensure the long-term survival of F.thunbergii.But the genetic diversity of F.thunbergii is relatively higher in population levels while lower at the species levels and the degree of genetic differentiation occured among the populations is not significant.The germplasm resources are relatively stable among these six populations.These populations could be used to breed the fine strains of F.thunbergii as the bases.

In order to make a report on 90 cases of insomnia treated by the Wei-defensive qi-regulating and brain-strengthening needling technique,as a clinical subject of the multiple center,for assessing the preliminary standardized indications and operating methods and making an initial summary for writing the draft of the technique operation.Methods:By ratio of 1:1,the patients in conformity with the criteria were randomly divided into the experimental group and control group.In the experimental group,45 cases were treated with the Wei-defensive qi-regulating and brain-strengthening needling technique on Baihui (GV 20),Dazhui(GV 14),Shenmai(BL 62),Zhaohai(KI 6),plus ear points.In the control group,45 cases were treated with routine acupuncture on Sishencong(Ex-HN 1),Shenmen (HT 7),Sanyinjiao(SP 6),etc.The therapeutic effects in the two groups were observed and assessed in comparison.Results:The results showed cure in 15 cases.remarkable effect in 27 cases,effect in 42 cases,failure in 6 cases,and the total eriective rate in 94.4% in 90 cases of insomnia.In the three centers,the statistic management was performed based upon PSQI integral and showed significant difference(P＜0.01),and no significant difference among various groups(P＞0.01).Conclusion:The therapeutic effect in the treatment of insomnia by Wei-defensive qi-regulating and brain-strengthening needling technique is better than the control group and the therapeutic effect and methods in various centers are stable.without any deviation.

Objective To study the causes and protection of intracranial aneurysms uncompletely embolized with micro-coils. Methods 47 cases of intracranial aneurysms were treated with micro-coils to embolize the aneurysaml sac via femoral artery approach to endovascular embolization.The aneurysms were located on AcoA in 19,on MCA in 7,on PcoA in 16 and on ICA in 5,2 aneurysms were foundin each of 2 cases. Results The uncompletely occuluded in 12 of them,aneurysms were completely occluded in 35,9 aneurysms wereoccluded to 95%,2 aneurysms were occluded to 90%,1 neurysms were occluded to 80%. Conclusion The wide neck intracranialaneurysms,hugeness intracranial aneurysms and ophidian,irregular intracranial aneurysms are of high rate of uncompletely occluded.Usingbasket-technique,remodel technique,vein rebuild technique and nibble subdivision endovascular embolization can decrease the rate ofuncompletely embolize intracranial aneurysm.

OBJECTIVETo study the triterpenoid saponins in Hemsleya chensnsis.

METHODThe chemical constituents were isolated and purified by various chromatographic methods and their structures were elucidated on the basis of spectral analysis.

RESULTSeven known triterpenoid saponins were isolated from the root of H. chensnsis and were identified as 3-O-beta-D-glucuropyranosyl-oleanol-icacid (1), 3-O-beta-D-glucopyra-noside-oleanolicacid-28-O-beta-D-glucopyranoside (2), 3-O-(6'-methylester)-beta-D-glucuropyranosyl olea- nolic acid-28-O-alpha-L-arabinopyranoside (3), 3-O-(6'-methylester)-beta-D- glucuropyranosyl- oleanolic acid-28-O-beta-D-mannupyranoside (4), 3-O-(6'-ethyl ester)-beta-D-glucuropyranosyl oleanolic acid-28-O-beta-D- glucopyranoside (5), 3-O-alpha-L-arabinopyranoside-(1-->3) -beta-D-glucuropyranosyl- oleanolic acid-28-O-beta-D-glucopyranoside (6), 3-O-beta-D-glucu-ropyranosyl-oleanolic acid-28-O-beta-D-glucopyra-noside-(1-->6)-beta-D-glucopyranoside (7).

CONCLUSIONCompounds 1-7 were isolated from this plant for the first time.

Cucurbitaceae ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Mitragyna ; chemistry ; Molecular Structure ; Plant Extracts ; pharmacology ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective:To investigate the effects of Danggui Shaoyao San(DSS) on cognitive function and neuronal apoptosis in vascular dementia (VD) rats.Methods:Fifty SPF grade male SD rats aged 6-7 weeks were randomly divided into sham operation group, model group, positive drug group (nimodipine group, 9.45 mg·kg -1), DSS low-dose group (1.6 g·kg -1), DSS high-dose group (6.4 g·kg -1) according to random number table, with 10 rats in each group. The VD rat model was established by permanent ligation of bilateral common carotid arteries. Seven days after modeling, the rats in different groups were administrated by gavage according to corresponding interventions, once a day, for 28 days. Morris water maze test was used to evaluate the learning and memory ability of rats.The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and reactive oxygen species (ROS) in hippocampal area of rat brain were detected by ELISA.The protein expressions of apoptosis-related proteins Bcl-2, Bax, cleaved Caspase-3 and leptin receptor/glycogen synthase kinase 3β microtubule-associated protein tau(LEP-R/GSK-3β/tau) signaling pathway were detected by Western blot. GraphPad Prism 9 software was used for statistical analysis of data, repeated measure ANOVA and one-way ANOVA were used for comparison between multiple groups, and SNK- q test was used for further pairwise comparison. Results:The results of water maze experiment showed that the time and group interaction of escape latency of the five groups were not significant ( F=1.223, P>0.05), the main effect of group and time were significant ( F=74.65, 18.32, both P<0.05). On the 5th day, the escape latency of nimodipine group, DSS low-dose group and DSS high-dose group were lower than that of model group ( q=14.425, 7.477, 21.392, all P<0.05), and that of DSS high-dose group was lower than that of nimodipine group ((15.28±2.46)s, (22.78±3.31)s, q=6.966, P<0.05). There was statistically significant difference in the number of crossing platforms of rats in 5 groups ( F=17.331, P<0.05). The numbers of platform crossing in nimodipine group and DSS high-dose group were higher than that in model group ( q=6.789, 10.635, 5.270, all P<0.05), and the number of platform crossing in DSS high-dose group was higher than that in nimodipine group ((6.84±1.63), (5.22±1.75), q=3.846, P<0.05). ELISA results showed that the levels of MDA, ROS and SOD in hippocampal tissues of rats in 5 groups were significantly different ( F=49.338, 38.518, 15.440, all P<0.05). The levels of MDA and ROS in hippocampus of DSS high-dose group were lower than those of model group ( q=16.061, 13.541, both P<0.05) and nimodipine group ( q=4.317, 5.162, both P<0.05), SOD level of DSS high-dose group was higher than those of model group ( q=8.179, P<0.05) and nimodipine group ( q=4.135, P<0.05). Western blot results showed that the levels of apoptosis-related proteins Bcl-2/Bax and Caspase-3 were significantly different in the 5 groups ( F=30.692, 43.384, both P<0.01). The level of Bcl-2/Bax in DSS high-dose group was higher than that in model group ( q=10.562, P<0.05) and nimodipine group ( q=3.820, P<0.05), the level of Caspase-3 was lower than those of model group ( q=12.139, P<0.05) and nimodipine group ( q=7.734, P<0.05). The levels of LEP-R, p-GSK-3β, p-S404 tau and p-S202 tau expression level in hippocampal tissues of the 5 group were significantly different ( F=80.927, 59.230, 159.784, 105.923, all P<0.01). The levels of LEP-R and p-GSK-3β protein in nimodpine group and DSS high-dose group were higher than those in model group ( q=16.275, 20.104, both P<0.05; q=12.942, 17.257, both P<0.05), the levels of p-S404 Tau and p-S202 Tau in the two groups were lower than those in model group ( q=19.121, 27.456, both P<0.05; q=17.559, 22.780, both P<0.05). The levels of LEP-R(0.98±0.15), (0.86±0.14)) and p-GSK-3β((0.95±0.16)s, (0.82±0.13)) in DSS high-dose group were higher than those in nimodipine group ( q=3.829, 4.314, both P<0.05), the levels of p-S404 Tau((0.41±0.03)s, (0.58±0.07)) and p-S202 Tau((0.48±0.05)s, (0.59±0.06)) in DSS high-dose group were lower than those of nimodipine group ( q=8.335, 5.220, both P<0.05). Conclusion:DSS can improve the cognitive function of VD rats, and the mechanism may be related with reducing oxidative stress level, inhibiting neuronal apoptosis, and upregulating LEP-R/GSK-3β/Tau signaling pathway.

Objective:To report 1 case of mushroom poisoning complicated with multiple organ dysfunction syndrome treated by liver transplantation, so as to explore surgical effectiveness and prognosis of liver transplantation on children mushroom poisoning.Methods:Retrospectively analyzed a child with mushroom poisoning who was admitted to the First Affiliated Hospital of Zhengzhou University in June 2017.Conventional treatment was not ideal, the child was conducted orthotopic liver transplantation.The effectiveness was evaluated and the feasibility of liver transplantation in treating children mushroom poisoning with literatures was reviewed.Results:A male 2-year old child obtained, preo-perative Child-pugh class for C. This child recovered well postoperatively, the jaundice disappeared, and liver enzyme and blood coagulation function improved significantly within 1 week after liver transplantation.However, the head magnetic resonance imaging presented brain atrophy and ventricular expansion, with intelligence and athletic ability being significantly behind his peers through postoperative regular follow-up for 2 years.Conclusion:Fulminant hepatic failure that was caused by mushroom poisoning is an indication for liver transplantation, and orthotopic liver transplantation is an effective method for the treatment of mushroom poisoning, which can effectively improve liver function, metabolic disorders and blood coagulation function for children with mushroom poisoning, but nervous system damage that was caused by mushroom poisoning is irreversible.Although liver transplantation technology successfully cured liver failure, it still left severe neurological sequelae.

OBJECTIVE: To compare the effect of 7 antipsychotic drugs on the life quality of schizophrenia patients including chlorpromazine, sulpiride, clozapine, risperidone, olanzapine, quetiapine, and aripiprazole. METHODS: A total of 1,227 stable schizophrenic patients within 5 years onset who took 1 of the 7 study medications as maintenance treatment were followed up for 1 year at 10 China sites. Patients were evaluated by the short form-36 health survey (SF-36) at the baseline and at the end of 1 year. RESULTS: The life quality was improved obviously at the end of the follow-up. There was significant difference in body pain, vitality, and mental health (P<0.05) among these antipsychotic drugs. CONCLUSION All 7 antipsychotic drugs can improve the life quality of schizophrenia patients. Atypical antipsychotic drugs, especially olazapine and quetiapine, are superior to typical antipsychotic drugs in improving life quality.
Adolescent ; Adult ; Antipsychotic Agents ; therapeutic use ; Benzodiazepines ; therapeutic use ; Dibenzothiazepines ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Olanzapine ; Quality of Life ; Quetiapine Fumarate ; Schizophrenia ; drug therapy ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo observe the clinical efficacy differences between acupuncture combined with 40-min penetration moxibustion and 60-min penetration moxibustion at back-points for insomnia of insufficiency of heart and spleen type.

METHODSSixty patients of insomnia with insufficiency of heart and spleen type were randomly assigned into a 40-min group and a 60-min group. The two groups were treated with acupuncture at Jueyinshu (BL 14), Xinshu (BL 15), Geshu (BL 17), Pishu (BL 20), Shendao (GV 11) and Zhiyang (GV 9). With moxibustion box, the penetration moxibustion was applied at the back until sweating and redness on the back. The moxibustion was given for 40 min in the 40-min group and 60 min in the 60-min group. The treatment was given once a day, five days per week. Each session was consisted of 5 treatments, with an interval of 2 days between session and totally 4 consecutive weeks were provided. The Pittsburgh sleep quality index (PSQI), TCM symptom scale were observed and recorded before and after treatment in the two groups. The even temperature at raising period, effective period, reducing period, as well as minimum high temperature, comfortable temperature, minimum cold temperature and medication status were compared; also the effect was compared between the two groups.

RESULTSThe total effective rate was 96.6% (28/29) in the 60-min group, which was higher than 89.3% (25/28) in the 40-min group (<0.05). Compared before treatment, the total score of PSQI and sleep quality, sleep time, sleep efficiency, sleep disorder, daytime dysfunction as well as the total TCM symptom score and its drowsiress in the morning, palpitation, amnesia, appetite were reduced after treatment in the 40-min group (all<0.05). After treatment, the total score and each score of PSQI as well as total score and each score of TCM symptom scale were reduced after treatment in the 60-min group (all<0.05). After treatment, the total score and each score of PSQI as well as total score and each score of TCM symptom scale were significantly different between the two groups (all<0.05).

CONCLUSIONSAcupuncture combined with penetration moxibustion can improve the symptomsof insomnia with insufficiency of heart and spleen type, which is more significant in the 60-min group, indicating prolonged time of penetration moxibustion can improve sleep latency.

The penetrating moxibustion technique is proposed based on experience of the ancients and clinical practice for many years. From the aspects of definition, action characteristics and technique at different parts, the advantage and application experience of penetrating moxibustion have been discussed. The adequate dose of moxibustion is necessary in penetrating moxibustion; in addition, moxibustion sensation should be penetrating; therefore, with a temperature of 43℃ for more than 20 min, sweating, flushing, speckle appearing after penetrating moxibustion. Due to individual differences of age, gender and constitution factors, the effects of penetrating moxibustion are different, in clinical practice the body reaction and patient's feelings should be deliberately focused on other than does or sore and blister of moxibustion. The technique is common used in the abdomen, waist and knee joint, etc.