Objective Severe preeclampsia (sPE), which is usually complicated by small-for-gestational-age (SGA) and immature labor , remains a leading cause of maternal and neonatal mortality and morbidity. This study was to investigate the risk factors of SGA in sPE. Methods We retrospectively analyzed 100 cases of sPE, 35 with SGA (the case group) and the other 65 without it ( the control group ) .We conducted single-factor analysis on the general characteristics and potential impact factors of the patients , i-dentified the independent risk factors of SGA using the un-conditional stepwise logistic model , and assessed the value of umbilical arter-y S/D ratio and proteinuria ration in the diagnosis of sPE with SGA with the ROC curve . Results Compared with the control group , the case group had more cases of early-onset sPE ( P =0.010 ), earlier gestational and delivery weeks (P<0.001), lower neonatal weight at birth ( P<0 .001 ) , higher rate of admission to and longer stay in the neonatal ICU (P<0.001), and higher incidence of neo-natal complications (P<0.05).The case group also showed signifi-cant increases in comparison with the control in the umbilical artery S/D ratio (2.95 ±0.75 vs 2.31 ±0.47, P<0.05), frequency of S/D ratio ≥95th percentile (22.86% vs 6.15%, P<0.01), and rate of proteinuria ≥5 g/24 h (42.86% vs 20.00%, P<0.05).The S/D ratio ≥95th percentile (OR=6.02, 95%CI:2.32-16.78) and proteinuria≥5 g/24 h (OR=1.65, 95%CI:1.56-3.01) were found to be the risk factors of sPE with SGA.The area under the curve was 0.852 for the combination of S/D ra-tio and proteinuria ration in the diagnosis of sPE with SGA (P<0.05). Conclusion Umbilical artery S/D ratio≥95th percentile and proteinuria ≥5 g/24 h contribute to the early prediction, prevention, and prognosis of sPE, and is valuable for the diagnosis of sPE with SGA.

Objective:To investigate the current status of cognitive frailty among elderly patients in Urumqi and to identify its influencing factors.Methods:From March to December 2019, the elderly from 3 tertiary hospitals′ geriatrics centers in Urumqi were recruited using the general information questionnaire, FRAIL Scale, Montreal Cognitive Assessment and Clinical Dementia Rating.Results:A total of 1 006 elderly patients were surveyed, among which, 131(13.0%) cases were deemed to have developed cognitive frailty. The results of multivariate logistic regression analysis showed that age, depression, Instrumental Activities of Daily Living Scale score and diabetes were influencing factors of cognitive frailty ( P<0.001). Conclusion:The prevalence rate of cognitive frailty in elderly patients is relatively high. Medical staff should attach great importance to the assessment of cognitive frailty in elderly patients and take targeted intervention in time to prevent, slow down or reverse the onset and development of cognitive frailty.

Objective: To evaluate the clinical efficacy of sinew-regulating bone-setting manipulations plus exercise therapy in treating chronic non-specific low back pain (CNLBP). Methods: A total of 65 CNLBP patients were divided into two groups by the random number table method. Thirty-three cases in the treatment group were intervened by sinew-regulating bone-setting manipulations plus exercise therapy; 32 cases in the control group were intervened by medium-frequency electrotherapy plus exercise therapy. Before and after treatment, visual analog scale (VAS), dynamic and static muscle endurance of low back, median frequency (MF) of surface electromyography (sEMG) and Oswestry disability index (ODI) were used to evaluate the low back function. The therapeutic efficacy was estimated after treatment. Results: The two groups each had 2 dropouts during the study. The total effective rate was 90.3％ in the treatment group versus 66.7％ in the control group, and the between-group difference was statistically significant (P<0.05). After treatment, the VAS score, dynamic and static muscle endurance of low back, MF of sEMG and ODI score all changed significantly in both groups (all P<0.05); all the items in the treatment group were significantly different from those in the control group (all P<0.05). Conclusion: Sinew-regulating bone-setting manipulations plus exercise therapy can effectively release pain in CNLBP patients, increase muscle endurance of the low back and improve the quality of life, and its therapeutic efficacy is more significant than that of medium-frequency electrotherapy plus exercise therapy.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

AIM: To prepare and purify the polyclonal antibodies against human myofibrillogenesis regulator 1 (hMR-1), then to characterize the purity, titer, specificity and the availability.METHODS: Two polypeptides named peptide 1 and 2 were synthesized based on the bioinformatics analysis of the sequence of hMR-1 by using software TMHMM and DNAStar, then coupled with keyhole limpet hemocyanin (KLH) for immunization. These peptides for immunization were mixed and injected into New Zealand rabbits to prepare antibodies specifically against hMR-1. ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, antibodies were identified by Western blotting and immunocytofluorescent assays. Applications of the antibodies on neonatal rat cardiomyocytes were also employed.RESULTS: (1)The titers of antibodies were 1:10~5. In WB assay, a specific 17kD band was detected, corresponding to the predicted molecular weight of hMR-1; the positive fluorescent signals were distinct. (2)On the neonatal rat cardiomyocytes model, we observed a peri-nucleus location. The fluorescent signal of hMR-1 overexpression group was much stronger than that in vector control and normal control groups.CONCLUSION: All these results indicate that the antibodies obtained from poly peptides mixture immunization have either human original or rat original antigens. The antibody is available for using in Western blotting or immunofluorescent assays.

Objective To detect γ-secretase gene mutations in a large Chinese pedigree with acne inversa (AI).Methods Clinical evaluation was carried out in a large pedigree with AI through field investigation.Peripheral blood samples were obtained from 17 family members (11 affected and 6 unaffected) and 100 unrelated healthy human controls.DNA was extracted from the blood samples,and PCR was performed to amplify all the coding regions of PSEN 1,PSENEN and NCSTN genes followed by DNA sequencing analysis.Results There were 67 members over 5 generations in this family,of whom,25 (13 males and 12 females) were affected by AI.AI was inherited in an autosomal dominant manner in this family.Skin lesions were mainly distributed on the neck,back,chest and buttocks,and occasionally in subaxillary regions.DNA sequencing revealed a novel missense mutation,c.1258C＞ T (p.Q420XP),in the exon 11 of the NCSTN gene in 11 affected family members,which leads to a substitution of glutamine by a premature termination codon at amino acid 420 (p.Q420X).The mutation was undetected in either the unaffected members or the unrelated healthy controls,and had not been registered in the single nucleotide polymorphism (SNP) database in National Center for Biotechnology Information.Conclusions There is a novel heterozygous missense mutation,c.1258C ＞ T in the exon 11 of the NCSTN gene,which may be the molecular basis of pathogenesis of AI in this family.

Objective To express human chemokine-like factor 1 (CKLF1) in Drosophila S2 cells and study its function. Methods The pMT/V5-His-CKLF1 expression plasmid was constructed and transfected into Drosophila S2 cells. The positive clones were selected through PCR and RT-PCR. The culture medium was analyzed by Western blot with anti-CKLF1 polyclonal antibody. Chemotaxis and MTT assays on human peripheral blood and C2C12 cells, respectively, were then carried out with the medium. Results CKLF1 was transcribed efficiently in S2 cells. The expressed CKLF1 protein could be detected in the culture supernatant by Western blot, which showed weak chemotactic activity on both human peripheral blood neutrophils and lymphocytes as well as enhancing effect on the proliferation of C2C12 cells. Conclusion CKLF1 was expressed successfully in Drosophila S2 cells and secreted into the culture medium. The recombinant CKLF1 expressed in Drosophila cells can chemoattract leucocytes and promote the proliferation of C2C12 cells.

Objective:To isolate endophytic fungi from Panax ginseng and study their antifungal and antitumor activities.Methods:Endophytic fungi were isolated from 5-year-old garden ginseng and 15-year-old transplanted ginseng.The antifungal active endophytes were screened with Pyricularia oryzae P-2b model using mircodilution method,and the activities of endophytes against pathogenic fungi were tested in vitro.The antitumor activities of the endophytes were examined by MTT method in vitro.Results:Sixteen(33.3%)of the 48 endophytic fungi fully suppressed the activity of P.oryzae P-2b;11(22.9%)colonies showed satisfactory antifungal activities against Candida albicans,Cryptococcus neoformans,Trichophyton rubrum,and Aspergillus fumigatus;and 5(10.4%)colonies showed satisfactory antitumor activities against tumor cell lines MKN45,LOVO,HepG2,and HL-60.Among the bioactive colonies,Yuan-25 showed best antifungal activity,with its MIC80 aganinst Trichophyton rubrum being 4 mg/L,which was similar to that of fluconazole.Yuan-27 showed the best antitumor activity,with its IC50 similar to that of doxorubicin.Conclusion:Isolated endophytic fungi of Panax ginseng has antifungal and antitumor activities and is worth further exploring.

Objective:To assay aflatoxin B1 in the oil as a pharmaceutical excipient in soft capsules by LC-MS/MS. Methods:Aflatoxin B1 was extracted from the peanut oil in soft capsules by the solvent composed of methanol and 0. 1% formic acid solution, and then centrifuged and the supernatant was purified by neutral alumina cartridges and tested after the concentration with the mobile phase consisting of methanol and 0. 1% formic acid solution with gradient elution at the flow rate of 0. 3 ml·min-1 . 25μl of the tested solu-tion was injected for the analysis at the column temperature of 30℃. Electrospray ionization ( ESI) source was applied and operated in the position ion mode. Multiple reactions monitoring ( MRM) mode was used to quantify the samples. Results:Aflatoxin B1 was in good linearity within the range of 0. 098-1. 960 μg·L-1(r=0. 999 5). The limit of detection was 0. 05 μg·L-1. The average sampling recovery was 97. 73% (n=6) with RSD of 4. 625%. Conclusion:The method is proved to be sensitive, accurate, specified and re-producible, which is referential for the assay of aflatoxin B1 in oily preparations.

Background The conventional drugs for preventing and treating graft rejection have the risks of inducing adverse responses.Researches showed that resolvinE1 (RvE1) can regulate Th1 cell-mediated immunoreaction.However,whether RvE1 has an inhibit effect on high-risk corneal graft is unclear now.Objective This study was to investigate the effects of RvE1 on immune rejection in high-risk corneal grafting mouse models.Methods SPF BALB/c mice were used as recipients,C57BL/6 mice were as donors.Ninety BALB/c mice were divided into corneal allograft group,corneal allograft+RvE1 group and corneal autograft group according to random number table.High-risk corneal graft models were established by corneal suturing for 14 days and followed by penetrating keratoplasty in recipients.Allograft keratoplasty was performed on the right eyes in the mice of corneal allograft group and corneal allograft+RvE1 group,and self-corneal graft rotated 180°was transplanted on the right eyes in the mice of autograft group.Normal saline solution of 10 μl was subconjunctivally injected after surgery once per day for 7 days in the corneal allograft group and corneal autograft group,and 10 μl RyE1 (1 μg) was used in the same way in the corneal allograft+RvE1 group.The recipient eyes were examined for potential rejection signals with slit lamp microscope and calculated the mean survival time and rejection index (RI).The histopathology was examined 21 days after modeling by hemotoxylin and eosin staining.The expressions of CD4 and interferon-γ (IFN-γ)in the corneas were detected by immunohistochemistry.Th1 cell (CD3+CD8a-IFN-γ+) percentage in draining lymph nodes were measured by flow cytometry.The mRNA expression levels of interleukin-2 (IL-2),tumor mecrosis factor-α (TNF-α),IFN-γ and T-bet were detected by real-time fluorescence quantitative PCR.Results The mean survival time of grafts was (28.5± 1.7) days in the corneal allograft group,and that in the corneal allograft+RvE1 group was (14.0±1.6) days,showing a significant difference between them (t =4.14,P＜0.001),while the survival rate was 100％ at 50 days after modeling in the corneal autograft group.Corneal edema and inflammatory cell infiltration were slight in the corneal allograft+RvE1 group and corneal autograft group compared with corneal allograft group.CD4 was positively expressed in corneal tissue,and IFN-γ was expressed in corneal epithelium.The CD4+ and IFN-γ+ cell number was decreased in the corneal allograft+RvE1 group and corneal autograft group compared with corneal allograft group under the fluorescence microscope.The percentages of Th1 cells in lymph cells of corneal allograft +RvE1 group and corneal autograft group were (1.07 ±0.25) ％,(0.85 ±0.12) ％,respectively,which were significnatly lower than (1.56±0.20) ％ in the corneal allograft group (both at P＜0.05).The expressions of IL-2,TNF-α,IFN-γ and T-bet mRNA in the corneal tissue in the corneal allograft group were higher than those in the corneal allograft+RvE1 group and corneal autograft group (all at P＜0.05).Conclusions RyE1 inhibits graft rejection in high-risk allograft mouse models probably by down-regulating the Th1 cell percentage in lymph cells and the expression of inflammationrelated cytokines in corneal grafts.