Objective: Asphyxia and ventricular fibrillation are the two most prevalent causes of cardiac arrest. The study investigated the differences in brain damage after cardiac arrest between asphyxial and ventricular fibrillation cardiac arrests in rats. Methods: Male healthy Sprague-Dawley rats were randomly assigned to the asphyxial group (cardiac arrest of 6 min, n=15), ventricular fibrillation group (cardiac arrest of 6 min, n=15) and sham group (n=5). Neurologic deficit scores and tape removal test were evaluated at 1, 3 and 7 days after cardiopulmonary resuscitation from three groups. Serum S-100B and brain histopathologic damage scores were also examined. Results: There were no differences in neurologic performance at 1, 3 and 7 days after cardiopulmonary resuscitation between the asphyxial group and ventricular fibrillation group (P>0.05, respectively). Serum S-100B level was higher in the asphyxial group at 1, 3 and 7 days, compared with the ventricular fibrillation group (P<0.05, respectively). There were significantly higher histopathologic damage scores at 1, 3 and 7 days in the asphyxial group compared with the ventricular fibrillation group (P<0.05, respectively). Conclusion: Asphyxial cardiac arrest has worse morphologic brain damage compared with ventricular fibrillation cardiac arrest, but the functional brain damage caused by asphyxial cardiac arrest is similar to that caused by ventricular fibrillation cardiac arrest.

Objective To design the small interference RNA (siRNA) specific to human CCL20 gene by RNA interfering technique, construct its recombinant lentiviral expression vectors, and identify these vectors by DNA sequencing. Methods According to Tuschl’s principle, the siRNA was designed and converted into cDNA of shRNA (small hairpin RNA) of siRNA for CCL20 gene. The cDNA was synthesized and inserted into plasmid pHSER-dsRNA-GFP-SIN which was linearized by restriction endonucleases SpeⅠ and SalⅠ. The recombinant plasmid was transformed into competent E. coli. DH5? cells. The positive recombinant colony was selected by ampicillin medium agar and identified by DNA sequencing. Results Two recombinant lentiviral plasmids of siRNA specific to CCL20 gene were constructed successfully. Their DNA sequence analysis completely coincided with their designed sequences. Conclusion Lentiviral vector-based siRNA expression plasmids against CCL20 gene have been successfully constructed and identified. They will be further used for interfering CCL20 mRNA transcription and lay the foundation for CCL20 gene modified human keratinocyte stem cells.

BACKGROUND:Donkey-hide glue reinforcing bone oral solution (DGRBOS) is effective on preventing and treating fracture, but the mechanism of harmacology is still not clear. Vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF-2) are important cytokines, which can promote blood vessel growth and osseous anabolism during fracture healing.OBJECTIVE: To investigate the effects of DGRBOS on expression of VEGF and FGF-2 in the process of fracture healing of SD rats' fracture of tibia in bony callus, and explore the mechanism of DGRBOS in the treatment of fracture.DESIGN: A completely randomized controlled study.SETTING: Department of Traumatic Orthopedics, Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: Ninety female Sprague-Dawley (SD) rats, with a mean weight of (368±40) g, aged about 3 months, wereprovided by Center of Animal Experiment, Tongji Medical College, Huazhong University of Science and Technology.Experimental drug: DGRBOS was donated by Xinjiang Huashidan Pharmaceutical Co., Ltd. The qili linking bone pill,positive control drug, was purchased from Hunan Pharmaceutical Co., Ltd.METHODS: This experiment was carried out in the Laboratory for Bone Metabolism of Integration of Chinese and Western Medicine (Laboratory of Provincial Level), Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from June to November 2005. Transverse midshaft fractures were produced on the right tibia in these rats using three-point bending technique, and 90 rats were randomly divided into three experimental groups:the DGRBOS group (n =30): according to medicine conversional method between human being and animal, 2 mL DGRBOS was fed to every mouse by intragastric administration at 2 vices/day; the qili linking bone pill group (positive control group, n =30): the pills dissolved in distilled water at 225 g/L, and consequently were administered intragastrically into mice at 2 mL/vice and 2 vices/day; normal saline group (negative control group, n =30): normal saline was administered intragastrically into mice at the coordinative capacity and same frequences. Selecting 4, 7, 14,21 and 28 days during the experiment, the immunohistochemistry method was adopted to detect the change of expression of VEGF and FGF-2 through computing value of mean optical degree (MOD) and number of the positive cells.

OBJECTIVETo establish and evaluate chronic heart failure (CHF) rat model of Xin-qi deficiency complicated blood stasis and edema syndrome (XQD-BSES).

METHODSTotally 40 SD rats were randomly divided into the normal control group (Control), the propylthiouracil (PTU) group, the adriamycin (ADR), and the ADR + PTU group. Normal saline was used as equivalent solvent of each group. Rats in the Control group were intragastrically and intraperitoneally injected with normal saline. Rats in the PTU group were intragastrically injected with PTU suspension and intraperitoneally injected with normal saline. Rats in the ADR group were intragastrically injected with ADR solution and intraperitoneally injected with normal saline. And rats in the ADR + PTU group were intragastrically injected with PTU suspension and intraperitoneally injected with ADR solution. The dose of PTU was 0.2% of daily forage weight, once daily. The dose of ADR was 3.5 mg/kg, once per week. The modeling lasted for 6 weeks. Left ventricular ejection fraction (LVEF), left ventricular fraction shortening (LVFS), brain natriuretic peptide (BNP), heart rate (HR), respiratory rate (RR), urine output, ear temperature, exhaustive swimming test (EST), Tri-iodothyronine (T3), tetra-iodothyronine(T4), thyroid stimulating hormone (TSH) as well as heart, lung, liver weight indices and their pathological sections were integrated and compared.

RESULTSCompared with the Control group, LVEF, LVFS, BNP, HR, RR, heart, lung, liver weight indices, urine output, ear temperature, EST, and T3, T4, and TSH changed significantly in the ADR group, the PTU group, and the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure occurred in pathological sections of heart, lung, and liver. Compared with the ADR group, LVEF, LVFS, BNP, and lung, liver weight indices, urine output, ear temperature, T3, T4, and TSH changed significantly in the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure were more serious in pathological sections of heart, lung, and liver. Compared with the PTU group, LVEF, LVFS, BNP, HR, RR, urine output, EST, T4, heart and lung weight indices changed significantly in the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure were quite serious in pathological sections of heart, lung, and liver.

CONCLUSIONADR + PTU was an appropriate method to establish CHF rat model of XQD-BSES.

Animals ; Edema ; Heart Failure ; diagnosis ; Heart Ventricles ; Humans ; Judgment ; Models, Animal ; Qi ; Rats ; Ventricular Function, Left

OBJECTIVETo study clinical outcomes of Chinese medidine fumigation and massage therapy for the treatment of knee stability and functional recovery after anterior cruciate ligament reconstruction operation,and to explore the effect on tendon-bone healing.

METHODSTotal 50 patients were divided into two groups: the control group (normal rehabilitation therapy group),the treatment group (Chinese medicine fumigation and manipulation group). There were 25 patients in the control group, including 16 males and 9 females, who were treated with isometric muscle training, with the gradually enlarging amplitude of flexion and progressive loading of bearing training for knee recovery. There were 25 patients in the treatment group, including 15 males and 10 females,who were treated with the conventional rehabilitation therapy combined with Chinese medicine fumigation and massage therapy. The Chinese herbs named as Haitongpi decoction was steamed by a special equipment to fumigate the knee after operation; Based on the biomechanical parameters of the ligament reconstruction, the massage therapy was designed to control the degree of the knee flexion and release the adhesion for early recovery of knee functions. The Lysholm knee function evaluation system was used, and MRI examination was performed to measure the change in width of ligament tunnel in femur and tibia to evaluate the safety and stability of the treatment.

RESULTSLysholm system showed that two groups both had improving results from the 1st month after operation to the 3rd month (treatment group, F=36.54, P<0.05; the control group, F=28.12, P<0.05), and the results of the treatment group was better than that of the control group at the observation point (the 1st month, t=0.105, P<0.05; the 3rd month, t=5.361, P<0.01). There was no difference between the two groups when evaluating the bone and tendon healing 3 and 12 months after operation (P>0.05), indicating that Chinese rehabilitation therapy was a safety treatment without the influence on the loosing of tendon.

CONCLUSIONChinese medicine fumigation and massage therapy can early improve the knee function after the anterior cruciate ligament reconstruction operation without the disturbance of the knee stability.

Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Anterior Cruciate Ligament Reconstruction ; Case-Control Studies ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Fumigation ; Humans ; Knee Injuries ; drug therapy ; physiopathology ; surgery ; therapy ; Knee Joint ; drug effects ; physiopathology ; surgery ; Male ; Massage ; Range of Motion, Articular ; Recovery of Function

OBJECTIVETo study and evaluate the curative effect and mechanism of Qiangxin Granule (QXG) in intervening chronic heart failure (CHF) rats with Xin-qi deficiency complicated blood stasis and edema syndrome (XQD-BS-ES).

METHODSTotally 72 SD rats of clean grade were randomly divided to the normal control group (n =10) and the model group (n = 62). The XQD-BS-ES rat model was established by adriamycin plus propylthiouracil method. Survived modeled rats were then randomly divided to 5 groups i.e., the model group (n = 11, administered with normal saline by gastrogavage), the Western medicine (WM) group (n =11 , administered with perindopril and hydrochlorothiazide by gastrogavage), the low dose QXG (QXG(L)) group (n = 11, administered with 9.26 g/kg QXG by gastrogavage), the middle dose QXG (QXG(M)) group (n = 11, administered with 18.52 g/kg QXG by gastrogavage), the high dose QXG (QXG(H)) group (n = 11, administered with 37.04 g/kg QXG by gastrogavage). After 4 weeks of treatment, left ventricular ejection fraction (LVEF), left ventricular fraction shortening (LVFS), brain natriuretic peptide (BNP), heart rate (HR), respiratory rate (RR), urine output, ear temperature, exhaustive swimming test (EST), tri-iodothyronine (T3), tetra-iodothyronine (T4), thyroid stimulating hormone (TSH), as well as heart, lung, liver weight index and their pathological sections, and high sensitivity C-reactive protein (HS-CRP), angiotensin II (Ang II), carbohydrate antigen 125 (CA125) were detected and compared.

RESULTSCompared with the normal control group, LVEF, LVFS, BNP, HR, RR, urine output, ear temperature, EST, T3, T4, TSH, HS-CRP, Ang II, and CA125 changed significantly in the model group (P < 0.01). Compared with the model group after treatment, LVEF, LVFS, BNP, urine output, EST, T4, heart and liver weight index, HS-CRP, Ang II, CA125 were significantly improved in each QXG group (P < 0.05, P < 0.01). Moreover, TSH was improved in the QXGL and QXG(M) groups (P < 0.05); ear temperature and T3 in the QXG(M) were also improved (P < 0.05); the lung weight index decreased in the QXG(M) and QXG(H) groups (P < 0.01). Compared with the WM group, T4 and CA125 were obviously improved in all QXG groups (P < 0.01); BNP and ear temperature were obviously improved in QXG(L) and QXG(M) groups (P < 0.05, P < 0.01); LVEF, LVFS and TSH were obviously improved in the QXG(M) group (P < 0.05, P < 0.01). And as far as each treatment group, LVEF, LVFS, urine output increased significantly after treatment (P < 0.01); EST obviously increased in QXG(M) and QXG(H) groups (P < 0.01); ear temperature increased in all QXG groups (P < 0.05, P < 0.01). Moreover, compared with the model group, pathological changes of heart, lung, and liver were improved to some degree in each treatment group, especially in the QXG(M) group.

CONCLUSIONSGood curative effect was shown in each QXG group. QXG could improve LVEF, LVFS and BNP of CHF rats of XQD-BS-ES, as well as T3, T4, TSH, EST, urine output, and ear temperature. Moreover, QXG showed superiority than WM group in this respect.

Angiotensin II ; Animals ; C-Reactive Protein ; Chronic Disease ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Edema ; Heart ; Heart Failure ; drug therapy ; Heart Ventricles ; Medicine, Chinese Traditional ; Natriuretic Peptide, Brain ; Qi ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Thyrotropin ; Ventricular Function, Left

Objective:To compare the clinical and prognostic characteristics of ovarian endometrioid carcinoma (OEC) patients with synchronous endometrial lesions and patients with pure OEC.Methods:A retrospective review of the medical records of patients received initial treatment and a postoperative pathological diagnosis of OEC at Peking University People′s Hospital between August 1998 and December 2017 were performed. According to the inclusion criteria, a total of 56 patients with OEC were included in the study, including 13 patients concurrent with simultaneous endometrial lesions (Group A) and 43 patients with pure OEC (Group B).Results:Patients with synchronous endometrial lesions accounted for 23% (13/56). Mean age of Group A at diagnosis was (44.9±8.3) years old, 2/13 of patients were postmenopausal, and no one had a history of hypertension, the first symptom of 5/13 people was irregular vaginal bleeding. Mean age of Group B patients at diagnosis was (52.7±10.2) years old, 53% (23/43) of patients were postmenopausal, and 28% (12/43) patients had the history of hypertension, the first symptom of 4 (9%, 4/43) people was irregular vaginal bleeding. The differences of age, menopause status, history of hypertension and initial symptoms between the two groups were statistically significant (all P<0.05). There were no significant differences in fertility history, dysmenorrhea history, age of menarche, history of endometriosis, preoperative and postoperative CA 125 level, International Federation of Gynecology and Obstetrics (FIGO) stage, tumor grade, metastatic site and platinum-based chemotherapy drug resistance between the two groups (all P>0.05). The overall 5-year survival rate of OEC patients was 91.6%, and the overall 5-year progression-free survival rate was 76.6%. Among them, the 5-year survival rate of the OEC concurrent with simultaneous endometrial lesions group was 80.2%, and the pure OEC group was 93.4%; the 5-year progression-free survival rate of the OEC concurrent with simultaneous endometrial lesions group was 74.1%, and the 5-year progression-free survival rate of the pure OEC group was 77.3%. There were no significant differences between the two groups (all P>0.05). Multivariate analysis showed that the independent factors for the prognosis of OEC patients were FIGO stage ( P=0.006) and residual lesion size ( P=0.020). Conclusions:OEC patients have a high proportion of simultaneous endometrial lesions. OEC with simultaneous endometrial lesions are younger than patients with pure OEC. Synchronous endometrial lesions do not affect the prognosis of patients with OEC.

OBJECTIVE: To establish a new method of SNP typing. METHODS: Based on the principle of allele specific PCR and capillary electrophoresis technique, 11 diallelic SNP loci were selected and two forward primers with different length were designed for each SNP, with their 3' ends matched to the two alleles, respectively. An artificially mismatched base was also introduced into the third or fourth base in the 3' end area of the two forward primers in order to enhance the specificity of amplification. A common reverse primer was designed 100-300 bp away from the forward primers, and labeled with fluorescence. The PCR products were separated and analyzed by ABI Prism 310 Genetic Analyzer after all of the 11 SNPs were multiply amplified. RESULTS: A single product peak was observed while the SNP was homozygous, and two product peaks with different height were observed while the SNP was heterozygous. The length of PCR products was different with the different SNPs. According to the length of the products and the number of the product peaks, the genotypes of the 11 SNPs can simultaneously be analyzed, and the results were in accordance with the direct sequencing. CONCLUSION Fragment length discrepant allele specific fluorescence labeled multi-PCR (FLDASFLM-PCR) is a simple, rapid and efficient new method for SNP typing.
Alleles ; Electrophoresis, Capillary/methods* ; Forensic Genetics ; Humans ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide/genetics*

OBJECTIVETo investigate the value of single nucleotide polymorphism array (SNP array) for the identification of de novo mutations in the DMD gene among fetuses.

METHODSG-banded karyotyping and SNP array were performed on a fetus with intrauterine growth restriction but without family history of Duchenne/Becker muscular dystrophy (DMD/BMD). Multiplex ligation-dependent probe amplification (MLPA) was subsequently applied on amniocytes and maternal peripheral blood sample to detect DMD gene deletion/duplication mutations.

RESULTSKaryotyping of amniocytes showed a normal 46, XY karyotype. SNP array on amniocytes detected a 116 kb deletion (chrX: 32 455 741-32 571 504) at Xp21.1 with breakpoints at introns 16 and 30 respectively, encompassing exons 17-29 of the DMD gene. In addition, MLPA analysis of the DMD gene on amniocytes confirmed the deletion of exons 17 to 29 identified by SNP array. However, no deletion/duplication mutation was detected by MLPA in the mother.

CONCLUSIONThe de novo deletion of exons 17 to 29 of the DMD gene detected in the fetus may result in BMD or DMD. SNP array can improve the efficiency for detecting genomic disorders in fetuses with unidentified pathogenic genes, negative family history and nonspecific phenotypes.

Adult ; Dystrophin ; genetics ; Exons ; genetics ; Female ; Fetus ; abnormalities ; Gene Deletion ; Humans ; Muscular Dystrophy, Duchenne ; genetics ; Phenotype ; Polymorphism, Single Nucleotide ; genetics ; Pregnancy

Two hundreds postmenopausal women with osteopenia, aged 45-80, were randomly divided into 4 groups, and received different drug interventions. After treatment for 12 months, the lumbar spine bone mineral density(BMD)and total hip BMD in alendronate group increased significantly(3.5% and 2.6%, both P＜0.05), the lumbar spine BMD raised 2.0% in selective estrogen receptor regulator group(P＜0.05). Bone turnover indices improved in both groups(all P＜0.05). No change in BMD or bone turnover indices was found in patients treated with active vitamin D3and calcium.