OBJECTIVETo observe the effect of Cangfu Congxian Decoction (CCD) on oxidative stress in granulosa cells of polycystic ovary syndrome (PCOS) patients.

METHODSForty PCOS patients underwent in vitro fertilization-embryo transfer (IVF-ET) were assigned to the treatment group and the control group 1 according to random digit table, 20 in each group. Patients in the treatment group took CCD (200 mL, once in the morning and once in the afternoon) 2 months before IVF-ET, while those in the control group 1 took no Chinese medical decoction. Recruited were another 20 patients undergoing IVF-ET for tubal factors (as the control group 2). The clinical effect of IVF-ET were observed, including oocyte retrieval number, 2 pronuclear (2PN) fertilization rate, good quality embryo rate, clinical pregnancy rate, and ovarian hyperstimulation syndrome (OHSS) induced transplantation cancel rate. The expression of relative oxygen species (ROS) in granulosa cells was detected using cell immunofluorescence combined with confocal microscopy and FCM.

RESULTSCompared with the control group 1, occyte retrieval number, 2PN fertilization rate, and good quality embryo rate increased in the control group 2 and the treatment group (P <0. 05). OHSS induced transplantation cancel rate decreased in the control group 2 (P < 0.05). Fluorescence intensity of ROS decreased in the treatment group and the control group 2, as compared with the control group 1 (P < 0.01).

CONCLUSIONCCD increased good quality embryo rate by down-regulating the expression of ROS protein in ovarian granulosa cells, and correcting in vivo oxidative stress.

Drugs, Chinese Herbal ; therapeutic use ; Embryo Transfer ; Female ; Fertilization in Vitro ; Granulosa Cells ; drug effects ; metabolism ; Humans ; Oocyte Retrieval ; Ovarian Hyperstimulation Syndrome ; prevention & control ; Oxidative Stress ; drug effects ; Polycystic Ovary Syndrome ; drug therapy ; Pregnancy ; Pregnancy Rate ; Reactive Oxygen Species ; metabolism

Objective To analysis the immunological characteristics of patients with severe H1N1 influenza A, and to provide theoretical basis for predicting the prognosis of the disease. Methods A retrospective analysis was conducted. The clinical data of 15 patients diagnosed with severe H1N1 influenza A and admitted to Shanghai General Hospital of Nanjing Medical University from October 2015 to December 2016 were collected. All the patients were divided into survival and death groups according to 28-day survival. Clinical characteristics, treatment algorithm, organ function, inflammatory reaction and immune cell status were compared, and Cox regression was used to decide the risk factors of 28-day death in patients with severe H1N1 infection A. Results All 15 patients with severe H1N1 infection A were enrolled, most of who presented with cough (93.3%), fever (86.7%), sputum production (80.0%), shortness of breath (73.3%), myalgia (40.0%) and fatigue (40.0%). All had been received anti-virus, antibiotics, mechanical ventilation and anti-coagulation therapy; some were treated with prone position, neuromuscular blocker and extracorporeal membrane oxygenation (ECMO). The incidences of acute myocardial and kidney injury were high, and the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score (14.1±6.1) and sequential organ failure assessment (SOFA) score (9.6±4.1) implicated the critical condition. Of 15 patients, 4 patients died in 28 days, while 11 were cured and discharged. Compared with survival group, the patients in death group had higher levels of APACHE Ⅱscore (22.7±3.8 vs. 11.8±3.8), troponin [cTn (μg/L): 0.52 (0.07, 2.02) vs. 0.15 (0.10, 0.45)] and blood urea nitrogen [BUN (mmol/L): 11.9 (6.7, 29.1) vs. 3.9 (2.7, 6.8)] and a lower level of blood platelets count [PLT (×109/L): 76±33 vs. 146±49, all P < 0.05]. The levels of C-reactive protein (CRP) and interleukin-6 (IL-6) within 24 hours of admission in death group were significantly higher than those of survival group [CRP (mg/L): 172.2±88.5 vs. 74.8±33.1, IL-6 (ng/L):283.3 (140.1, 711.0) vs. 18.5 (12.7, 71.4), both P < 0.01]. Compared with survival group, the expressions of CD3+, CD4+, CD8+ T cells and natural killer cell (NK cell) in death group were significantly decreased (CD3+ T cell: 0.348±0.119 vs. 0.573±0.106, CD4+ T cell: 0.135±0.046 vs. 0.344±0.098, CD8+ T cell: 0.089±0.057 vs. 0.208±0.054, NK cell: 0.124±0.057 vs. 0.252±0.182, all P < 0.05), but there were no significant differences in CD4+/CD8+ ratio and human leucocyte antigen-DR positive (HLA-DR+) T cell between death group and survival group (CD4+/CD8+ ratio:1.57±0.26 vs. 1.83±0.54, HLA-DR+ T cell: 0.035±0.022 vs. 0.062±0.036, both P > 0.05). B lymphocyte in death group was significantly higher than that of survival group (0.477±0.136 vs. 0.229±0.121, P < 0.01). Cox regression analysis revealed that APACHE Ⅱ score [risk ratio (RR) = 20.4, 95% confidence interval (95%CI) = 5.3-31.2, P = 0.017], CD4+ T cell (RR = 11.1, 95%CI = 5.1-20.0, P = 0.048) and CD8+ T cell (RR = 9.1, 95%CI = 4.3-16.7, P = 0.049) were independently risk factors of 28-day survival of patients with severe H1N1 influenza A. Conclusion Immunological paralysis and severe inflammatory response were early complicated with severe H1N1 influenza A, and these were significantly associated with prognosis.

Objective To investegate the effect of ginsenoside Rg1 on the apoptosis related protein FLICE-inhibitory protein(FLIP),Fas-associated death domain protein (FADD)and Caspase-3 in the subatania nigra(SN)of 1-methyl-4-phenyl-1,2,3,6-tetrahyd-ropyridine (MPTP)-induced mouse models of Parkinson’s disease(PD), and to investigate the role of FADD and FLIP in the pathogenesis of PD and the protective effect of ginsenosides Rg1 on dopaminergic neurons.Methods 45 C57BL/6N mice were randomly divided into control group,model group and ginsenoside Rg1 group (n=15).The mice in model group were injected with MPTP by intraperitoneal,the mice in Rg1 group were injected with ginsenoside Rg1 before injecting MPTP,and the mice in control group were injected with normal saline by intraperitoneal. The behavioral changes of the mice in various groups were observed, and immunohistochemistry and Western blotting methods were used to observe the expressions of tyrosine hydroxylase (TH),FADD,FLIP and Caspase-3 in substantia nigra of the mice.Results Compared with control group,the mice in model group presented with typical symptoms of PD, the TH-positive neurons in the subatania nigra was significantly reduced (P<0.01 ), the number of FADD, FLIP and Caspase-3 positive cells was significantly increased(P<0.01),and the cytoplasm was deeply stained;the protein expression levels of FADD,FLIP and Caspase-3 were significantly increased (P<0.01).Compared with model group,the PD symptoms of the mice in ginsenoside Rg1 group reduced, the number of TH-positive neurons was significantly increased, the number of positive cells of FLIP,FADD and Caspase-3 were significantly reduced(P<0.01),and the cytoplasm was lightly stained;the protein expression levels of FADD, FLIP and Caspase-3 were significantly reduced (P<0.01 ). Nonlinear correlation analysis found that there was a positive relationship between the number of FADD and Caspase-3 positive cells (r=0.791,P<0.05).Conclusion Ginsenoside Rg1 may play a neural protective effect dopaminergic on neurons by modulating the FADD and FLIP expressions in SN of PD model mice.

Objective To study the clinical,cardiopulmonary functional and hemodynamic profiles of systemic sclerosis patients with pulmonary hypertension (SSc-PAH) compared with those of idiopathic pulmonary hypertension (IPAH).Methods Patients diagnosed with SSc-PAH or IPAH by right heart catheterization were consecutively enrolled into the study between 2011 and 2013 in Peking Union Medical College Hospital (PUMCH).Cases with pulmonary hypertension related to other diseases were excluded.Demographic data,laboratory parameters,6 minutes walk distance (6MWD),pulmonary function and hemodynamic variables at the time of diagnosis were collected and compared between the two groups.Results A total of 20 SSc-PAH patients including 19 females and one male with age of (43.1 ± 12.2) years,and 18IPAH patients including 16 females and 2 males with age of (38.4 ± 12.4) years were enrolled in this study.Subjects in both groups had similar mean pulmonary arterial pressure,cardiac index and pulmonary vascular resistance (PVR) when recruited.Compared with IPAH patients,SSc-PAH patients showed significantly decreased all parameters including forced vital capacity (FVC)％ [(77.1 ± 13.2)％ vs (88.6 ± 14.9)％,P =0.026],diffusing capacity of the lung for carbon monoxide (DLCO) ％ [(46.2 ±13.1) ％ vs (66.6± 13.3)％,P ＜0.001],DLCO/alveolar ventilation(VA) [(55.1 ± 14.3)％ vs (75.1 ± 11.5)％,P ＜0.001],and 6MWD [(365.6 ±85.1) m vs (454.3 ± 136.8) m,P =0.034].In subgroup analysis of SSc-PAH patients,elevated PVR (OR 2.122,95％ CI 1.093-4.119,P =0.026) and decreased DLCO％ (OR 0.916,95％ CI 0.842-0.996,P =0.040) were independently associated with reduced 6MWD.Conclusions Under the similar hemodynamic condition,SSc-PAH patients had more severe restrictive ventilation dysfunction and diffusion capacity dysfunction.Decreased 6MWD in SSc-PAHpatients was probably related to the impairment of pulmonary function.

Objective Toevaluatea modified Ziehl-Neelsen(Z-N) stain in the diagnosis of tuberculous meningitis. Methods Cerebrospinal fluid specimens from 35 patients were stained by using the modified Ziehl-Neelsen staining. Re-sults The positive rate was 94.29% in 35 patients with tuberculous meningitisand the intracellular acid-fast bacilli was detected in 53.40%of all specimens. One case was stained positive in 15 patients with non-tuberculous meningitis. Con-clusion The modified Ziehl-Neelsen stain not only significantly improves the detection rates of tuberculous meningitisbut alsois able to identify intracellular M.tuberculosisin cerebrospinal fluidspecimen.Thus, the modified Z-N stain can be a convenient tool for diagnosing tuberculous meningitis.

Objective To evaluate the rehabilitation nursing care of double endo-button technique under total arthroscope for acute acromioclavicular joint dislocation.Methods From August 2007 to March 2009,8 cases (8 shoulders,6 males and 2 females,22 - 53 years old,mean age 34.4 ) with acute acromioclavicular joint dislocation ( Rockwood type Ⅲ) were treated in author' s hospital.After surgery we made systemic rehabilitation care plan,phased into the body functional rehabilitation exercises,under the premise of ensuring safety of patients as soon as possible to urge the active and passive functional exercise,regular follow-up.Radiographic analysis and the Constant scores and VAS ( Visual Analogue Scale) were used to provide a final evaluation of shoulder function for 3 months,6 months and 12 months after operation.Results All of 8 patients were followed up for more than 3 months( average 7.2 months).The Constant score averaged 93.9,with a mean VAS score of 1.75.There were no patients with A-C joint re-dislocation during this period after analyzing X-ray,but VAS score of 1 patient was 4 by 3 months.The operation for 8 cases was satisfactory.Most patients returned to their pre-injury activity level.Conclusions Total sxthrosocopic double endo-button technique for reconstruction of coracoclavicular ligament is an effective approach to treatment of fresh Rockwood Ⅲ acromioclavicular dislocation. This approach enjoys earlier recovery and less complication. Postoperative rehabilitation nursing is as important as surgery,not only for good functional recovery,but also for patient satisfaction.

OBJECTIVETo observe the effect of Bushen Wenyang Huayu Recipe (BWHR) on hypoxia inducible factor-1α (HIF-1α), proline hydroxylase2 (PHD2), von Hippel Lindau disease (VHL) suppressor gene expressions in endometriosis (EM) rats with Shen yang deficiency blood stasis syndrome (SYDBSS), and to explore the pathogenesis of EM and the mechanism of BWHR for treating EM.

METHODSTotally 50 SD rats were randomly divided into five groups, i.e., the blank control group, the sham-operation group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 in each group. Rats in the blank control group and the sham-operation group were fed routinely. Rats in the rest 3 groups received 30-day "extended refrigerator freezing and ice water immersion" and combined with " autotransplantation" to establish EM rat model with SYDBSS. One Milliliter BWHR at 3.33 g/mL was administered to rats in the CM group by gastrogavage. Gestrinone at the daily dose of 0. 5 mg/kg was administered to rats in the WM group by gastrogavage. Equal volume of normal saline was administered to rats in the model group, the blank control group, and the sham-operation group. The size and morphology of ectopic foci in rats were observed after 4 weeks of medication. Expressions of serum CA125, plasma cyclic adenosine monophosphate (cAMP), and plasma cyclic guanosine monophosphate (cGMP) were detected by radioimmunoassay. Morphological changes of eutopic endometrium and ectopic tissue were observed under the optical microscope by HE staining. Protein expressions and contents of HIF-lα, PHD2, and VHL were detected by immunohistochemical SABC method and Western blot. mRNA expressions of HIF-1α, PHD2, and VHL were detected by RT-PCR.

RESULTSThe ectopic foci grew significantly in the model group. Their volumes were obviously contracted after treated by CM and WM. Compared with the blank control group and the sham-operation group, serum CA125 and plasma cGMP obviously increased, cAMP obviously decreased (P < 0.05); expressions and contents of HIF-1α mRNA and protein all decreased (P < 0.05); mRNA and protein expressions and contents of PHD2 and VHL all decreased in the model group (P < 0.05). Compared with model group, levels of CA125 and cGMP obviously decreased; cAMP levels obviously increased, expressions and contents of HIF-1α mRNA and protein all increased, mRNA and protein expressions and contents of PHD2 and VHL all increased in the WM group and the CM group (P < 0.05). Compared with the CM group, PHD2 protein contents were higher in the WM group (P < 0.05). HIF-1α was negatively correlated with PHD2 (r = -0.799, P = 0.00). HIF-1α was negatively correlated with VHL (r = -0. 625, P = 0.003).

CONCLUSIONSBWHR could effectively treat EM. Its mechanism might be associated with reducing contents of HIF-1α, serum CA125, and plasma cGMP, and up-regulating expressions of PHD2, VHL, and cAMP.

Animals ; Cyclic AMP ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; drug therapy ; metabolism ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Proline ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Up-Regulation ; Yang Deficiency ; drug therapy ; metabolism

ObjectiveTo investigate distingwished clinical and experimental characteristics of the four main subtypes in myelodysplastic/myeloproliferative neoplasms (MDS/MPNs).MethodsMDS/MPNs 53 cases from Provincial Hospital Affiliated to Shandong University,including 24 cases CMML,13 cases aCML,12 cases JMML,4 cases MDS/MPN-U,were analyzed regarding to 2001 WHO classification.Morphology (M) of peripheral and bone marrow blood cells were observed under microscope.FCM was used in immunological(Ⅰ) analyse on blasts and myelomonocytes in peripheral blood and bone marrow.G-banding technique was used in cytogenetic (C)examination.PCR was used in molecular genetic (M) mutation detection.Numeric data,such as mean Hb,WBC,PLT et al,among several groups,were compared using Single-factor analysisof variance.Student-Newman-Keulstestwasuseincomparingmeansof two groups.Proportions,such as percentage of clinical features,immunological and cytogenetic abnormal cases among different groups,were compared using Chi-square test or Fisher exact test.Results( 1 ) In the course of MDS/MPNs,there were 46 cases (86.8％ ) had paleness and fatigue 33 cases (62.5％ ) had palpable spleen.JMML had most fever and enlargement of lymph node (75.0％,75.0％ ),statistically distinguished from CMML ( 12.5％,12.5％ ) (x2 =14.89,17.98,P ＜ 0.05 ).(2) The hemoglobin was ( 83.1 ± 24.6 ) g/L.WBC counts were ( 19.8 ± 8.1 ) × 109/L.PLT counts were ( 158.7 ± 108.2) x 109/L.Immature neutrophils and blasts were found in peripheral blood.(3)JMML and CMML had most monocytes absolute counts among the subtypes (4.25 ±0.76) (3.62 ±0.76).(4) Almost 100％ JMML had monocytes abnormalities.(5)For 15 cases were detected immunological characteres by FCM,13 cases showed abnormalities.(6)For 29 cases of MDS/MPNs had been analyzed chromosome karyotypes and 12 out of them (41.4％) were abnormal,Ph chromosomes and those AML-defining translocations were all negative,+ 8 and 7-involved- karyotypes were more frequent.(7)23 cases were detected molecular genetic features,in which were all negative.BCR/ABL1 and JAK2 V617F mutation were all negative in the 13 cases of aCML.JAK2 V617F mutation was positive in 1 case of MDS/MPN-U.ConclusionsMost MDS/MPNs had paleness and fatigue,light to mild anemia,cytosis,monocytes low grade of blast and immature neutrophils in peripheral blood with dysplasia in bone marrow.JMML seems has more severe clinical features and more distinguishing laboratory characters.Immunological abnormalities and abnormalkaryotypes are found frequently in MDS/MPNs with no statistical differences among the four subtypes.There is no specific molecular abnormals in MDS/MPNs.( Chin J Lab Med,2012,35:832-837)

Objective To study the molecular mechanisms of ampicillin- resistant haemophilius influenzae (Hi)in Nanjing. Methods One hundred and fifty- eight strains of Hi isolated from children were collected to detect bata-lactamase. TEM and ROB bata- lacta-mase genes were detected by polymerase chain reaction (PCR) ,and cloned into T vector for sequencing. Results The rate of ampicillin resistance was 41. 77% in Hi isolated from children in Nanjing,40.51 % was found to be bata-lactamase production. Eighty-nine strain were TEM positives, 1 strain was ROB positive,63 strains bata - lactamase positive ampicillin- resistant Hi were identified. The resistance mechanism of ampicillin resistant Hi was production of bata - lactamase , mainly TEM - type enzyme. Two bata - lactamase negative ampicillin - resistant Hi were identified , predicts the other mechanisms of ampicillin - resistant Hi was occuered yet . One strain of non -TEM - type,and non - ROB - type bata - lactamase - producing Hi was identified. Conclusions Ampicillin - resisitant in Hi isolated from children in this region is challenging. TEM bata - lactamase is the principal mechanism of ampicillin - resistant of Hi.

Objective To investigate CT features on increased cerebral vascular density and its pathological mechanism in patients with cyanotic congenital heart disease(CCHD).MethodsPreoperative brain CT scan and clinical data in 82 patients suffering from CCHD were analyzed. According to the increased levels of vascular density,patients were divided into 4 groups:normal,mild,moderate and severe.Relationships between the increased levels of vascular density and Hb,RBC,HCT,as well as the degree of cyanosis,were studied.AVONA was carried out to test blood CT value of cerebral sinuses,Hb,RBC and HCT in different groups. Descriptive analysis and linear regression were adopted to study the correlation between blood CT value and Hb concentration.The relationship of increased vascular density todegrees of cyanosis was analysed by Spearman.Results Among 82 patients,12 patients ( 14.6％ ) werefound in the group of normal vascular density and 70 patients ( 85.4％ ) in the increased vascular density group.Among 70 patients with increased vascular density,22 patients (26.8％ ) with (55.4 ± 2.6) HU,(169 ±6)g/L of Hb,(5.8 ±0.3) × 1012/L of RBC and 0.51 ±0.03 of HCT,29 patients (35.4％) with (61.3 ± 2.9) HU,(209 ± 15 ) g/L,(7.1 ± 0.4) × 1012/L,0.66 ± 0.06 and 19 patients ( 23.2％ ) with ( 68.8 ± 4.2) HU,( 242 ± 23 ) g/L,( 8.3 ± 0.9 ) × 1012/L,0.78 ± 0.08 were observed in the mild,moderateand severe group,respectively.There were significant differences in distribution of blood CT value ( HU),Hb,RBC and HCT in different groups ( F =163.263,134.703,120.974,136.541 ;P ＜ 0.01 ).Blood CT value was positively correlated with Hb concentration ( r =0.98,P ＜ 0.01 ). Vascular density was also positively correlated with the degree of cyanosis ( r =0.86,P ＜ 0.01 ). Conclusions Cerebral vascular density of patients suffering from CCHD presented different levels of increases based on CT scan results due to rise of RBC stimulated by anoxia.The increased level of vascular density was positively correlated with blood Hb concentration,and also associated with RBC accunulation caused by abnormal blood circulation.Moreover,it was positively correlated with the degree of cyanosis.