OBJECTIVETo explore the mRNA expressions of IL-10, IL-18 and Fas in peripheral blood mononuclear cells (PBMCs) in systemic lupus erythematosus (SLE) patients of various Chinese medicine syndrome types in order to seek the objective indices for syndrome differentiation of SLE.

METHODSSLE patients were sorted into various syndrome types by Chinese medicine syndrome differentiation. In them, patients with the 4 often encountered types were selected, i.e. the qi-xue excessive heat type (Group A, 12 patients), the Pi-Shen yang-deficiency type (Group B, 19 patients), the qi-yin deficiency with blood stasis type (Group C, 23 patients), and the qi stagnation and blood stasis with gan stagnation type (Group D, 28 patients). The mRNA expressions of IL-10, IL-18 and Fas in PBMCs were detected by SYBR Green I real time PCR.

RESULTSThe mRNA expressions of IL-10, IL-18 and Fas in PBMC of all SLE patients (0.40 +/- 0.53, 1.41 +/- 1.93 and 0.33 +/- 0.70) were significantly higher than those in the normal control group (0.07 +/- 0.11, 0.24 +/- 0.25, 0.08 +/- 0.21, P<0.01, P<0.05), their levels in different syndrome types decreased in order of type A-->B-->C-->D. The levels of the three indices in patients of type A and B were obviously higher than those in patients of type C and D (P<0.01), while no significant difference was found between type A and B (P>0.05), and between type C and D (P>0.05).

CONCLUSIONThe mRNA expressions of IL-10, IL-18 and Fas in PBMCs could be taken as the objective indices for reference in syndrome differentiation of SLE.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-18 ; blood ; Leukocytes, Mononuclear ; metabolism ; Lupus Erythematosus, Systemic ; blood ; diagnosis ; therapy ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; RNA, Messenger ; genetics ; Yang Deficiency ; diagnosis ; Yin Deficiency ; diagnosis ; Young Adult ; fas Receptor ; blood

Abstract: To analyze the clinical, therapeutic and laboratory characteristics of disseminated cryptococcosis caused by Cryptococcus neoformans invading the blood stream in patient with liver cirrhosis and splenectomy. A 30-year-old male underwent splenectomy plus pericardial devascularization due to "splenomegaly and hypersplenism" in March in 2016. The patient had intermittent fever after operation for many times, and successively accompanied with back pain, left lower limb abscess and right hip pain. The highest body temperature was 39 ℃. CT and MRI revealed the lung lesion and multiple bone destruction. During that period, the effect of antibiotics was not good. On April 19th, 2017, Gram's stain, India ink stain, API 32C, Vitek 2 Compact, ribosomal ITS and IGS sequence analysis were performed to identify the strain isolated from the pus and blood stream. The serum of the patient was detected for cryptococcal antigen. Antifungal susceptibility test was used to determine drug sensitivity and minimum inhibitory concentration (MIC). The Cryptococcus neoformans isolated from fresh pus specimen showed a prominent, thick capsule after India ink stain. The colonies isolated from pus and blood stream were identified Cryptococcus neoformans using API 32C, Vitek 2 Compact, and sequence analysis of rDNA ITS and IGS. Cryptococcal capsule antigen was positive. The minimal inhibitory concentrations of 5-Flucytosine, amphotericin B, fluconazole, itriconazole, voriconazole against the isolate were <4 μg/mL, <0.5 μg/mL, 4 μg/mL, ≤0.25 μg/mL, 0.125 μg/mL respectively. The patient was initially treated with intravenous amphotericin B and flucytosine. After anti-Cryptococcus treatment for two months, the patient clinically improved, and the lesions were reduced on a follow-up CT scan. The patient made a full functional recovery after treatment for six months. Cryptococcosis has hidden onset, atypical clinical symptoms and lack of specificity. Blood stream is the main channel for Cryptococcus to spread and involve many organs of the whole body, including skin, bone and so on. Therefore, early use of blood culture to monitor blood flow dissemination, actively removing the primary focus and cutting off the infection route in time and carrying out effective anti-Cryptococcus treatment are conducive to the patient's early recovery.

Objective To explore the clinical,radiological findings and pathogenic factor of spondylometaphyseal dysplasia.Methods Five cases were reported and the relevant documents were studied retrospectively.Plain X-ray film was performed in all patients.Results There were 4 male and 1 female,age ranged from 4 to 15 years with average of 9 years.The main features of 5 cases included delayed bone age,stature short,short trunk,waddling gait noted,scoliosis in 2 cases,kyphoscoliosis in 1 case,severe genu valgum in 2 cases.The main X-ray appearance of 5 cases is multiple irregularities of long bones metaphyses associated with platyspondylia,epiphysis is normal.Type Ⅰ SMD in 2 cases,Type Ⅱ SMD in 1 case,Type Ⅲ SMD in 2 cases.Conclusion When we meet children with delayed bone age and stature short,and muhiple irregularities of long bones metaphyses associated with platyspondylia were seen in X-ray plain film.we should think about spondylometaphyseal dysplasia.

Aim To investigate the radio-protective effect of schisandrol B(Sol B)underlying mechanism in alleviating gastrointestinal(GI)toxicity and intesti-nal damage induced by irradiation(IR)exposure dur-ing radiotherapy against abdominal and pelvic malig-nancies or the nuclear accident.Methods The mouse intestinal injury model was established through 4Gy ir-radiation.Immunohistochemistry,immunofluorescence staining,ELISA were used to measure DNA damage,apoptosis,inflammatory reaction,and oxidative stress in the intestine of mice after irradiation.The protective effect of Sol B on intestines injury was evaluated,and the mechanism of the related oxidative stress and in-flammatory response pathway was discussed.Results Sol B significantly improved the survival rates of intes-tinal cells upon IR exposure.The accumulation of DNA damage,apoptosis rate,and inflammatory factors in intestinal tissues were significantly inhibited.The decreased levels of ZO-1 and occludin induced IR inju-ry were rescued by Sol B,indicating the improvement of intestinal structure.The radio-protective activities of Sol B were involved in the stimulation of Nrf2 and the subsequent ferroptosis surveillance.Conclusion Sol B demonstrates a promising agent to ease intestinal in-jury during radiotherapy or nuclear accidents,in which GPX4 related ferroptosis surveillance mediated by Nrf2 is involved in the radio-protective effect of Sol B.

BACKGROUNDRoutine anteroposterior radiographs of the acromioclavicular (AC) joint with or without weight bearing have limitations in demonstrating the AC joint. Transarticular fixation with Kirschner wire is a treatment choice for AC dislocations. However, percutaneous fixation of the AC joint is technically demanding. The C-arm fluoroscopy can be used as routine intraoperative guidance to facilitate this procedure. The current study aims to introduce new projections, the axial and tangential views of AC joint, to help evaluate the severity of the injury and facilitate the percutaneous procedure.

METHODSThree shoulder specimens were used to find the projection directions of the axial and tangential views of the AC joint by using the digital radiography (DR) unit. The axial and tangential views were taken of 20 adult volunteers by referencing the projection directions determined in the shoulder specimens. The angles showed on the DR system and the angles between the coronal plane of the body and the vertical plane of the flat panel detector (FPD) during taking these radiographs were recorded. The C-arm fluoroscopy unit was used to take the axial and tangential views referencing the angles measured on the DR system. Routine anteroposterior radiographs of the AC joint were taken on the volunteers. The minimal distances from the distal clavicle to the acromion were measured on both tangential and anteroposterior radiographs. The data was statistically analyzed.

RESULTSThe clear axial and tangential radiographs of AC joints of the volunteers were obtained using both DR and C-arm fluoroscopy units. The angles demonstrated on the DR window are (20.8 ± 2.4)° for male and (18.3 ± 2.3)° for female. During taking the axial views, the angles between the coronal plane of the body and vertical plane of FPD are (23.3 ± 3.2)° for male and (20.1 ± 2.4)° for female. During taking tangential views, the corresponding angles are (117.5 ± 3.7)° for male and (113.1 ± 3.3)° for female. On the tangential radiographs, the minimal distance from the distal clavicle to the acromion is (6.1 ± 1.2) mm, wider than the same measurement on the anteroposterior radiographs (P < 0.05). Statistical analyses showed no significant differences in the above-mentioned angles and the minimal distances between the left and right AC joints (P > 0.05). There were no significant differences in the above-mentioned angles between DR and C-arm fluoroscopy units (P > 0.05).

CONCLUSIONSThe axial and tangential radiographs of the AC joint can demonstrate the joint clearly and they can be easily obtained with both DR system and C-arm fluoroscopy unit in similar projection directions.

Acromioclavicular Joint ; diagnostic imaging ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Radiographic Image Enhancement

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

Histone lysine methyltransferase EZH2 has been reported to be frequently overexpressed in hepatocellular carcinoma (HCC) tissues and associated with hepatocarcinogenesis. However, the exact mechanism of EZH2 up-regulation in HCC has not been determined. In this study, we used murine hepatocyte AML12 cells to investigate the role of hepatitis B virus X protein (HBx) in regulating the expression of mEZH2. Western blot analysis demonstrated that the expression level of mEZH2 protein in AML12 cells was up-regulated by HBx in a dose-dependent manner. To further investigate the mechanism of mEZH2 overexpression, the 2500 bp regulatory sequence upstream from the first exon of the mEZH2 gene was amplified from AML12 genomic DNA and constructed into a luciferase reporter plasmid. The luciferase activity of the mEZH2 promoter significantly increased in AML12 cells co-transfected with HBx plasmid, and deleting the -486/-214 promoter region decreased HBx-induced mEZH2 promoter activation by nearly 50%. The -486/-214 region was then analyzed in the TRANSFAC 6.0 database and a typical E2F1-binding site was found. Mutation of this E2F1-binding site or knockdown of E2F1 expression by RNAi led to a dramatic decrease in HBx-induced activation of the mEZH2 promoter and mEZH2 overexpression in AML12 cells. These results provide evidence that HBx up-regulates mEZH2 expression by transactivating the mEZH2 promoter through E2F1 transcription factor, thereby providing new epigenetic evidence for the carcinogenic effect of HBx.
Animals ; Binding Sites ; Cell Line ; E2F1 Transcription Factor ; genetics ; Enhancer of Zeste Homolog 2 Protein ; Hepatocytes ; cytology ; metabolism ; virology ; Histone-Lysine N-Methyltransferase ; genetics ; metabolism ; Mice ; Plasmids ; Polycomb Repressive Complex 2 ; Promoter Regions, Genetic ; genetics ; RNA, Small Interfering ; genetics ; Trans-Activators ; genetics ; metabolism ; Transfection ; Up-Regulation

BACKGROUND: Wharton's Jelly-derived mesenchymal stem cells are relatively primitive stem cells that are ideal vectors for gene therapy. However, there is a lack of studies on the conditions for the electrotransfection of Wharton's Jelly-derived mesenchymal stem cells. Therefore, exploring the optimal conditions for the electrotransfection of Wharton's Jelly-derived mesenchymal stem cells occupies an important position. OBJECTIVE: To investigate the effects of different electroporation conditions on the transfection efficiency of Wharton's Jelly-derived mesenchymal stem cells, and to explore the optimal conditions for cell electroporation. METHODS: By controlling the transfection conditions such as voltage, pulse duration and cell status, EEV-EGFP plasmids were transfected into Wharton's Jelly-derived mesenchymal stem cells by electroporation under different conditions. Transfection efficiency was detected by flow cytometry. RESULTS AND CONCLUSION: (1) The transfection efficiency was intended to increase when the voltage ranged from 125 V to 150 V, and the maximum transfection efficiency was obtained when the voltage was 150 V. However, when the voltage was further increased to 170 V, the transfection efficiency began to decrease considerably. (2) The maximum transfection efficiency was obtained when the pulse duration was 5.0 ms, while it was certainly decreased when the pulse duration was 2.5 and 7.5 ms. (3) The transfection efficiency of the cells cultured under normoxia was higher than that under hypoxic culture. These findings reveal that normally cultured Wharton's Jelly-derived mesenchymal stem cells can achieve higher electroporation efficiency via two pulse sessions at a voltage of 150 V, pulse duration of 5.0 ms, and pulse interval time of 50 ms.

Aim To investigate the protective effect of dimethyl fumarate on spleen injury induced by gamma radiation in mice and the related mechanism. Methods C57BL/6 mice were randomly divided into the blank control group, radiation model group and DMF administration group, which were administered once at 12 h before irradiation and once at 0. 5 h, 12 h, 24 h and 48 h after irradiation. The 30-day survival rate, body weight and pathological injury of spleen were measured after a one-time total body irradiation of Co 7 rays (8 Gy). TUNEL staining was used to detect apoptosis of spleen cells. Enzyme-linked immunoassay ( ELISA) was applied to detect the contents of TNF-a, IL-1 p, IL-6, IL-18, NLRP3 and AIM2 in spleen. Western blot test and immunofluorescence staining test was employed to verify the changes of NLRP3 and AIM2 contents in spleen tissue after irradiation. Results DMF could obviously improve the survival rate of irradiated mice, improve the weight loss of irradiated mice, re-duce the pathological injury of spleen, and inhibit the apoptosis of spleen cells after irradiation. ELISA results showed that DMF could significantly inhibit the increase of spleen inflammatory cytokines TNF-a, IL-lp, IL-6, IL-18 and inflammasome components NL-RP3 and AIM2 induced by irradiation. Western blot and tissue immunofluorescence staining also confirmed that DMF could inhibit the increase of NLRP3 and AIM2 inflammasome protein levels caused by irradiation. Meanwhile, NLRP3 agonist and AIM2 agonist could antagonize the radiation protection effect of DMF on spleen cells. Conclusion DMF can ameliorate spleen injury of Co 7-ray injured mice, and its mechanism is closely related to NLRP3/AIM2 inflamma-somes, which can be used as a potential protective drug for radiation injury.

OBJECTIVETo study the newborn's Apgar score in 'one minute' and relevant factors.

METHODSOne year inpatient woman from a Maternal and Child Health Hospital of Anhui province were selected by cluster sampling method and newborn asphyxia situation was investigated using Apgar score and self-designed questionnaire.

RESULTSThe Apgar score in 'one minute' which marking 8 to 10, 4 to 7 and 0 to 3 were found in 1875 (73.78%), 426 (16.77%) and 240 infants (9.45%) respectively. The average Apgar score in 'one minute' and five minutes were (7.69 +/- 2.27) and (9.01 +/- 1.89) respectively, The Apgar score in 'one minute' was significantly correlated with that of five minutes (Pearson coefficient correlation r = 0.841, P = 0.00). Ordinal regression analysis revealed that parturient age (OR = 1.04), being farmer (OR = 2.22), parity (OR = 1.26), assistant vaginal delivery (OR = 4.93), caesarean section (OR = 1.95), pregnancy-induced hypertension syndrome (OR = 1.42), albuminuria in gestational period (OR = 1.44), newborn being male (OR = 1.23), low birth weight (OR = 2.94), inborn abnormality (OR = 12.12), premature birth (OR = 1.22) and complications of delivery (OR = 5.04) were risk factors while the number of years under study (OR = 0.91), prenatal check-up (OR = 0.48), body length of newborn infant (OR = 0.88) and single birth (OR = 0.57) were protective factors.

CONCLUSIONApgar score in 'one minute' of newborn infant was affected by several factors as stated above. Health care program in earlier period toward community parturient should be strengthened in order to discover and control high risk factors of duration of pregnancy in earlier period.

Apgar Score ; Asphyxia Neonatorum ; epidemiology ; Epidemiologic Studies ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Risk Factors ; Time Factors