Objective To explore clinical effects of laparoscopy applied in the female patients with infertility. Methods 468 cases of female infertility patients were diagnosed and treated by laparoscopy and 168 cases received follow-up survey. Results Among those infertility patients, the patients of normal pelvic cavity and the abnormal one were 49(10.47% ) and 419( 89.53% ). Among those patients of abnormal pelvic cavity .fallopian tube disease in 276 cases(65.87% ) was at the first place. The constituent ratio of endometriosis,ovary tumor,polycystic ovary,uterine myoma,female genital tract malformation and pelvic tuberculosis was 14. 80% ,5. 97% ,4. 30% ,3. 82% , 3. 34% and 1.91% .respectively. The patients of primary infertility and the secondary ones were 208 and 260 cases. In secondary infertility patients 69.23% (180/260) of patients were caused due to fallopian tube diseases,and the constituent ratio was higher than that in primary infertility 46.15% (96/208) (x2 = 14.05 ,P <0.05),while in primary infertility,the constituent ratio of endometriosis and normal pelvic cavity were 20.19% ,15.38% ,which were higher than that in secondary infertility respectively(x2 = 11. 78,11. 16, all P < 0.05). The pregnancy rate of 168 cases followed-up was 45.24% (76/168).47.37% ,84.21% pregnancy occurred within 6,12 months with laparoscopy. The shortest and longest time of pregnancy was 2 and 30 months, respectively. The median was 8months. Conclusion Laparoscopy playd an important role in diagnosis of infertility and improvement of pregnancy rate.

Objective To construct a genetically-stable double auxotrophic,in which the uracil and leucine were mutated,using the Candida parapolymorpha ATCC26012 as materials.Methods Based on the physical and genetic engineering methods,the chromosome of the C.parapolymorpha strain was modified,where the ura3 and leu2 genes were directly mutated,to obtain the uracil and leucine double auxotrophic strain.Then the constructed strain was identified by the analysis of its biological properties,such as genetic stability,the change of the genes,and the physiologic and biochemical characteristics.Results The uracil and leucine double auxotrophic strain is obtained by screening.The biological identification results show that the obtained strain is genetically stable and the targeted genes are directly altered.In addition,the physiologic and biochemical analyses indicate that the auxotrophic can utilize various kinds of carbon and nitrogen nutrient sources,and its growth is good.Conclusion The successful construction of double auxotrophic mutant strains facilitated the genetic studies on C.parapolymorpha to meet various investigational purposes.Moreover,the constructed auxotrophic strains can be applied as advantageous host cells to express multiple proteins/antigens simultaneously,which is of great significance in the development of vaccines.

Objective To investigate the effect of hemorrhagic transformation (HT) on the outcome of acute ischemic stroke.Methods One hundred and one patients with acute ischemic stroke were divided into cerebral infarction without HT,hemorrhagic infarction (HI),and parenchymal hematoma (PH) according to the CT and/or MRI findings.The modified Rankin Scale (mRS) was used to evaluate the outcome at 3 months after onset,and the good outcome was defined as mRS ≤ 2.The multivariate logistic regression analysis was used to identify the predictors of outcome in patients with acute ischemic stroke.Results There were 53 patients without HT,44 HI,and 4 PH,respectively in 101 patients with acute ischemic stroke.HI (OR =0.07,95% CI0.01-0.30;P ＜0.001),＜65 years (OR =52.94,95% CI9.12-307.27;P＜0.001),and NIHSS score ＜ 15 (OR= 0.63,95% CI 0.50-0.78;P＜0.001) were the independent predictors of good outcome for patients with acute ischemic stroke.Conclusions HI is one of the independent predictors of good outcome at 3 months after acute ischemic stroke.

Objective To evaluate the immunogenicity of the recombinant human papillomavirus type 68b (HPV68b) virus-like particles(VLPs)in a mouse model.Methods The L1 protein of HPV type 68b was successful expressed in the Hansenula polymorpha strain (NVSI-68b-1).Processes including purifi-cation and reconstitution were performed to achieve pure HPV 68b VLPs.The purity, morphology and immu-nogenicity of the purified HPV 68 b VLPs were further analyzed .The BALB/c mice were immunized with HPV68b VLPs formulated on aluminum adjuvant .Pseudovirus-neutralizing antibody ( PsV NAb) assay was performed to detect the neutralizing antibodies in serum samples .Results The HPV 68 b L1 VLPs were ob-tained as indicated by the results of SDS-PAGE, Western blot assay , HPLC, electron microscopy and dy-namic light scattering with a high purity of 95%.Transmission electron microscopy and dynamic light scat-tering analysis revealed that the HPV68b L1 VLPs resembled the native virus with an average particle diame-ter of 50 nm.High levels of HPV68b-neutralizing antibodies were detected in serum samples from the mice immunized with HPV68b L1 VLPs.Moreover, a cross-protective efficacy of HPV68b L1 VLPs for HPV68a was observed .Conclusion This study suggested that the recombinant HPV 68 b VLPs expressed in a Han-senula polymorpha strain might be used as a potential candidate for the development of HPV vaccine .

Objective To investigate the psychological status of diarrhea type irritable bowel syndrome (D-IBS) patients overlapped with functional dyspepsia (FD) and its relationship with the changes of brain-gut peptides and cytokines levels.Methods A total of 190 D-IBS FD overlapped patients and 30 healthy controls were selected and all of them completed the questionnaire.The adult psychological testing system was applied for the Symptom Checklist (SCL-90) assessment.Plasma levels of 5-hydroxytryptamine (5-HT),somatostatin (SS),vasoactive intestinal peptide (VIP),endothelin (ET),interleukin (IL)-10 and IL-12 were measured in 59 randomly selected patients.Single factor analysis of variance and LSD test were performed for statistical analysis.Results In D-IBS-FD overlapped patients,the plasma 5 HT levels of the D-IBS patients and the FD patients were (1906.83±676.88) μg/L and (1745.53±653.23) μg/L respectively,both of them were significant higher than those of control group [(1289.44±918.45) μg/L,LSD test,both P＜0.05].However the levels of VIP and IL-10 were significant lower than those of control group (LSD test,both P＜0.01).In D-IBS patients,the plasma level of SS was significantly lower than that of control group (LSD test,P=0.025); the level of ET was significantly higher than that of control group and FD patients (LSD test,both P＜0.05).In D-IBS-FD overlapped patients,the scores of anxiety,depression and opponent were higher than those of control group (F=5.114,4.555 and 10.285,all P＜0.05).The plasma VIP and SS levels of patients with anxiety or depression were lower than those of control group (LSD test,both P＜0.05).The plasma VIP level was lower than that of patients without psychological disorders (LSD test,P=0.005).The plasma ET level of patients with disease course over five years was significantly higher than that of patients with disease course less than one year or between one and five years (LSD test,both P＜0.05),and the IL-10 level was significantly lower than that of patients with disease course less than one year (LSD test,P=0.004).Conclusions There were sub-healthlated with disease course,psychological disorder,the increased plasma 5-HT level and the decreased IL-10 level.There was close correlation between the lower gastrointestinal symptoms and the decreased levels of VIP,SS and the increased level of ET.

Objective To analyze the genome sequencing of mumps virus strain 79 purified from working seeds plague and examine its phylogenetic relationship with wild type virus strain isolated in China,and to explore the efficacy of vaccine at molecular level.Methods Whole genome sequences of two substrains of S79(major and minor),were obtained with fragment amplification by RT-PCR.Genetic distances between S79 and strains identified in China were analyzed based on the phylogenic analysis of small hydrophobic protein(SH) sequences.Results The nucleotide homologies of two S79 substrains(major and minor),with a ratio of 2:5 during culture,with Jeryl Lynn(JL) strain were 99.7% and 100%,respectively.There were scattered non-homologous recombination between two substrains.The genetic distances between strain S79,which was genotype A,and wild type virus strain identified in China,which were genotype F,was 11.2% -20.0%.S79 live vaccine was composed of two substrains,major and minor component,which were highly similar to JL strain in their genome sequences,but different from JL in their ratios during culture.Conclusion Different from wild type virus strain identified in China,the genotype of S79 was A,and phylogenetically distant from other strains,which may account for the low efficacy of S79 live vaccine.The ratios of two substrains might also be of interest for further study of the vaccine protection and efficacy.

OBJECTIVETo investigate the effect of Acanthopanax senticosus saponins (ASS) on oxidative damage induced by hydrogen peroxide (H2O2 ) in cardiomyocytes.

METHODThe cardiomyocytes were induced to oxidative damaged by exposed to H2O2. We evaluated the level of oxidative injury through morphology change, the survival rate, the activity of lactate dehydrogenase (LDH) and the content of cellular malondialdehyde (MDA). The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and the content of reductive glutathione (GSH) of cardiomyocytes were also determined.

RESULTWhen the cardiomyocytes were exposed to H2O, (50, 100, 200 micromol x L(-1)) for deigned time, the percentage of survival cells was down significantly (P < 0.01 or P < 0.001), and the activity of LDH and the content of MDA were increased markedly (P < 0.05, P < 0.01 or P < 0.001). These results show that the cells were subjected to oxidative damage. Treatment with ASS (600 mg x L(-1)) prior to H2O2 exposure could increase cell viability significantly, lessen cardiomyocyte morphological damaged change, and decrease LDH activity (1687.40 +/- 97.51) U x mL(-1) in media and cellular MDA content (16.50 +/- 2.66) nmol x mg(-1) markedly (P < 0.01 and P < 0.05). Furthermore, the activities of SOD (89.55 +/- 6.93) U x mg(-1), GSH-Px (845.87 +/- 63.76) mU x mg(-1), CAT (93.07 +/- 10.40) U x mg(-1) and the content of GSH (8.91 +/- 1.06) micromol x mg(-1) of cardiomyocytes were also raised by 600 mg x L(-1) l ASS (P < 0.05).

CONCLUSIONTaken together, the study implicate that ASS protects cardiomyocytes against oxidative-stress injury induced by H2O2 through reduction of lipid peroxidation and enhancement of the activity of antioxidant defense.

Animals ; Animals, Newborn ; Catalase ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Eleutherococcus ; chemistry ; Female ; Glutathione Peroxidase ; metabolism ; Hydrogen Peroxide ; pharmacology ; Male ; Myocytes, Cardiac ; drug effects ; enzymology ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Wistar ; Saponins ; pharmacology ; Superoxide Dismutase ; metabolism

Objective To evaluate the clinical value of CT perfusion (CTP) imaging for providing quantitative information about angiogenesis in patients with lung carcinoma and investigate the correlation of CTP enhancement parameters and histological microvessel density (MVD) with lymphatic involvement in peripheral lung carcinoma. MethodsFifty-three patients with pathology-proved peripheral lung carcinoma underwent CT perfusion scan before operation. The enhancement parameters of CTP were calculated based on the time-density curves (TDC) of fist pass phase. All cases were classified into two groups according to pathologic results: tumor with and without lymph node involvement. Two-sample t test was used for the statistics. The ROC curve was used to assess the efficiency of the enhancement parameters of CT perfusion and MVD for predicting lymphatic involvement.Results Tumors with lymph node involvement had significantly higher value of MVD than those without lymph node involvement (64.69±16.34 and 42.67± 16.78, respectively,t=4.84,P<0.01). Tumors with lymph node involvement had significantly higher value of CTP enhancement parameters (PH, M/A, PV) than those without lymph node involvement [PH= (41.79±15.50) and (29.99±10.91) HU,M/A =0.24±0.09 and 0.15±0.06, PV=(2.14±1.09) and (1.27±0.53) ml·min~(-1)·ml~(-1), t=3.21,3.95, 3.66, P<0.01, respectively]. The CTP enhancement parameters (PH, M/A, PV) of lung cancer correlated positively with the MVD, the highest correlation coefficient was between the PV and MVD (r=0.716, P<0.01). MVD and PV had higher values for predicting lymph node involvement in ROC curve analysis.The sensitivity, specificity and accuracy for predicting lymph node involvement were 80.8%, 81.5% and 81.1% or 84.6% ,85.2% and 84.9% respectively if MVD>52/0.74 mm~2 or PV>1.52 ml·min~(-1)·ml~(-1). ConclusionThe CT perfusion PV and histological MVD have good correlation with lymph node involvement in peripheral lung carcinoma and are important predicting parameters before operation.

Objective:To investigate the role of adenosine diphosphate ribosylation factor 6 (Arf6) in the pathogenesis of endometriosis.Methods:Endometrial tissues were sampled from women who were hospitalized in the Affiliated Hospital of Medical School of Ningbo University and Ningbo Women and Children′s Hospital from November 2020 to May 2021 with endometriosis ( n=44, endometriosis group) and without endometriosis ( n=17, control group). The expression of Arf6 protein in the endometrial tissues was detected by western blot. Endometrial epithelial cells from both groups were primary cultured and the distribution of intracellular mitochondria was detected by immunofluorescence. The expression of Arf6 protein was down-regulated by small interference RNA (siRNA), the distribution of mitochondria in cells with decreased Arf6 protein expression was observed, and the expression of mitochondria-related proteins development and differentiation enhancing factor 1 (DDEF1, also called AMAP1), reactive oxygen species 1 (ROS1) and epithelial-mesenchymal transition (EMT)-related proteins E-cadherin, vimentin were detected. Transwell assay was used to detect the changes in the migration ability of the cells. Results:Compared with the control group, ectopic endometrial tissue of endometriosis group showed high expression of Arf6 protein (0.174±0.019 vs 0.423±0.033; t=29.630, P<0.01); and in ectopic endometrial epithelial cells, mitochondria were distributed near the edge of the cell membrane. While Arf6 expression was down-regulated by siRNA, the distribution of mitochondria in ectopic cells returned to natural, close to the control level. In addition, the expression levels of AMAP1 and ROS1 in ectopic cells after Arf6 protein knockdown were significantly decreased. Transwell assay results indicated that knockdown of Arf6 could reduce the migration ability of ectopic epithelial cells [migration cell count: (34.3±7.5) cells]; and immunofluorescence verified low expression of E-cadherin but high expression of vimentin in ectopic epithelial cells, whereas knockdown of Arf6 protein E-cadherin expression increased but vimentin expression decreased. Conclusions:High expression of Arf6 protein in ectopic endometrial epithelial cells leads to the distribution of mitochondria tending to membrane marginalization, while inducing EMT, which are involved in the mechanism of endoheterosis pathogenesis.

Cryptococcal meningitis is a central nervous system infection caused by cryptococcal invasion of meninges and (or) brain parenchyma. It is characterized by severe intracranial hypertension and brain parenchyma damage. This opportunistic infection commonly occurs among HIV infected individuals, organ transplant recipients, and other immunosuppressive situations, and otherwise immunologically normal hosts. The global burden of human immunodeficiency virus/acquired immunodeficiency syndrome has declined over the years, but cryptococcal meningitis still has a high morbidity and mortality. The risk groups, clinical presentation, imaging characteristics, diagnostic criteria and therapeutic issues were reviewed in this article.