Objective To investigate the therapeutic effect of Chinese herbal medicine for seminal delayed liquefaction(SDL).Methods Two hundred and ninety SDL patients were randomized into two groups.Group A(N=230)received oral use of yin-nourishing and blood-activating herbal medicine(mainly composed of Radix Ophiopogonis,Radix Scrophulariae,Fructus Ligustri Lucidi,Herba Ecliptae,Radix Salviae Miltiorrhizae,Flos Carthami,Rhizoma Sparganii,Eupolyphaga seu Steleophaga),and group B(N=60)received intramuscular injection of chymotrypsin and oral use of vitamin C and zinc gluconate.The therapeutic effect in the two groups were evaluated after treatment,and the changes of main parameters of seminal liquid as well as the seminal plasma contents of prostate specific antigen(PSA),acid phosphatase(AP),citric acid(CA)and zinc were observed before and after treatment.Results In group A,156 patients were cured,57 effective,17 ineffective and the total effective rate was 92.6%;in group B,21 patients were cured,17 effective,22 ineffective and the total effective rate was 63.3%.The Therapeutic effect was better in group A than that in group B(P

Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.

To explore the clinical manifestations, diagnosis and treatment of pulmonary thromboembolism associated with protein C (PC)/protein S (PS) deifciency. Two male patients 29 and 26 years old diagnosed with PC deifciency and/or PS deifciency were retrospectively analyzed and related literatures were reviewed. The most common symptoms were pain in the lower limbs with chest pain or decreased vision. Color dopper lfow imaging (CDFI) showed lower deep venous phlebothrombosis. Multislice CT angiography (CTA) revealed pulmonary embolism. The level of serum homocysteine (HCY) increased and the level of plasma PC/PS content decreased to PC 57.4%, and PS 28.9%in patient 1, while PS 33.4%in patient 2. Poor routine anticoagulant response was observed. After the diagnosis of PC/PS deficiency, vitiamin B6 and B12 anticoagulant therapy was added, and the symptoms in the patients improved significantly. Congenital thrombophilia should be taken into consideration for young patients with lower deep venous thrombosis and pulmonary embolism which occur recurrently without obvious predisposing causes before 40. Plasma PC/PS concentrations or activity help a lot in the diagnosis and treatment.

Objective To investigate the guidance of the endoscopic ultrasonography for the surgery selec-tion of patients with low rectal cancerthrough analyzing the accuracy of endoscopic ultrasonography in preoperative TN staging. Methods Eighty-seven cases with low rectal cancer received preoperative endoscopic ultrasonography examination,the preoperative staging and the postoperative pathologic comparison. The EUS accuracy of preopera-tive staging of rectal cancer was evaluated. Results The preoperative staging with endoscopic ultrasonography for patients with low rectal cancer,100% in T1 stage,96.0% in T2 stage,85.7% inT3 stage,and 100% in T4 stage. The preoperative staging and the postoperative pathologic comparison in T stage were consistent(Kappa = 0.903, P < 0.05). The preoperative staging with endoscopic ultrasonography for patients with low rectal cancer ,87.0% in N0 stage,78.6% in N1 stage,and 100% in N2. The preoperative and the postoperative pathologic comparisons in N stage were consistent(Kappa = 0.768,P < 0.05). Conclusion The endoscopic ultrasonography had a certain advantage in the clinical preoperative evaluation for patients with low rectal cancer ,especially for invasion depth and the judgment of lymph node metastasis ,with a higher accuracy.

Objective:To investigate the roles and regulation mechanism of miR-31 in human cutaneous squamous cell carcino-ma (cSCC) growth. Methods:cSCC cells were transfected with the antisense oligonucleotide (ASO) of miR-31, and the cSCC growth was tested by colony formation and in vivo tumor formation assays. The target gene of miR-31 was validated by Western blot and green fluorescent protein (GFP) reporter assay. The cells were then transfected with the siRNA of the target gene, and the effect of the target gene on cell growth was preformed by colony formation assay. Finally, real-time PCR and immunohistochemistry were used for analy-sis of the expression of miR-31 and its target gene. Results:miR-31 ASO resulted in a low number of cell colonies and small tumor vol-ume (P<0.05). Western blot showed that the cells with miR-31 ASO had a higher protein level of large tumor suppressor homolog 2 (LATS2) than the control. The 3' UTR of LATS2 had a binding site with miR-31, and miR-31 ASO increased the GFP intensity con-trolled by LATS2 3' UTR, whereas no effect was observed on the mutant LATS2 3' UTR. Western blot showed that LATS2 siRNA inhib-ited the expression of LATS2 protein by about 80%. Knocking down of LATS2 increased the colony number by about 70%or 1.3-fold in cSCC cells. Real-time PCR showed that miR-31 was overexpressed in most cSCC tissues, compared with normal tissues. An inverse relationship existed between miR-31 and LATS2 expression levels. Immunohistochemistry validated that LATS2 was downregulated in cSCC tissues. Conclusion:miR-31, which functions as an oncogene, promotes cSCC growth by suppressing LATS2 expression. Our da-ta suggest that miR-31 is a potential miRNA-based therapeutic target for cSCC growth.

Objective To evaluate the application of sentinel lymph node biopsy in surgical operation for clinical stage N0 differentiated thyroid carcinoma.Methods One hundred and six patients who were diagnosed clinical stage N0 differentiated thyroid carcinoma in Department of Mammary Gland Thyroid Surgery of Qingdao Women and Children's Hospital between Jan.2012 and Jan.2013 were reviewed retrospectively.Carbon nanoparticles were used to trace sentinel lymph node in the surgery,the patients were divided into the dissection group (46 cases included 2 cases whose sentinel lymph nodes were not detected) and the non dissection group (60 cases) based on the rapid pathological results.The operation index,complications incidence and short-term outcomes were compared.Result The operative time,intraoperative blood loss and the faulty excision of parathyroid gland in dissection group is much higher than those in non dissection group (P ＜0.05).The incidence of provisionality hypoparathyroidism in dissection group is much higher than those in non dissection group (P ＜ 0.05).There were no permanent hypoparathyroidism and damage of laryngeal nerve in the two groups.Postoperative outpatient follow up was 30 months,no significant differences were seen in local regional recurrence rates(6.5％ vs 10％) between the two groups (P ＞ 0.05).There were no distant metastasis occurred in the two groups.Conclusions Lymph node metastasis can be found by sentinel lymph node biopsy,it is helpful for us to decide whether dissect central lymph node or not.Carbon nanoparticles is safe,which is helpful to protect the parathyroid gland.

Objective]To discuss the effect of sciatic nerve injury on the expressions of tumor necrosis factor-alpha(TNF-α), interleukin-1β(IL-1β)and interleukin-10(IL-10)in anterior cingulate cortex(ACC),and further to explain their roles resided in the development of neuropathic pain.[Method]With use of the methods of behavioral test,western blot and immunohistochemistry, we examine the effects of spared sciatic nerve injury(SNI)on the expressions of TNF-α,IL-1β,and IL-10 in ACC,and observe the effect of the neutralizing antibody of TNF-α,IL-1β on the rat mechanical allodynia.[Result]In present experiment ,SNI increased the protein levels of TNF-α,IL-10,but not IL-1β in ACC. Increased TNF-α-IR and IL-10-IR in ACC is located in neurons ,but not astrocytes and microglia at 7 d following L5-VRT. Pre-treatment with anti-TNFα antibody but not anti-IL-1βantibody into ACC significantly increased the rat paw withdrawal threshold to von Frey hairs.[Conclusion]These data suggested that the increased TNF-αin ACC neurons might be responsible for the development of neuropathic pain.

Objective To investigate the role of the p38 MAPK pathway in the formation of cytoplasmic vacuoles .Methods Af-ter treated with Anisomycin ,SB203580 or SP600125 ,images of HepG2 ,LM3 ,QBC939 ,Hela and A549 cells were recorded by light microscopy and taken at a magnification of 400 × .The effects of anisomycin ,SB203580 and SP600125 on the activity of p38 and JNK were measured by Western blot .LM3 and A549 cells were stained with the ER-tracker red and the lyso-tracker red and subjec-ted to confocal microscopy analysis .Results (1)Anisomycin could abolish cytoplasmic vacuolization of HepG2 cells .(2)p38 MAPK activation was responsible for anisomycin-induced cytoplasmic vacuolization abolishment .(3)p38 MAPK blocking initiated cytoplas-mic vacuoles formation in various cancer cell lines .(4)p38 MAPK blocking-induced cytoplasmic vacuoles disrupted the integrity of endoplasmic reticulum .(5)p38 MAPK blocking reversibly induced cytoplasmic vacuoles formation .Conclusion These observations provide direct evidence for a role of p38 MAPK signaling in regulating the formation of cytoplasmic vacuoles .

Background: Recent studies showed that the clinical outcome of moderately severe acute pancreatitis (MSAP) are different among different subgroups. Aims: To further subdivide MSAP, and explore the heterogeneity of MSAP subgroups. Methods: A retrospective analysis was performed on patients with acute pancreatitis (AP) from January 2016 to December 2020 at Northern Jiangsu People’s Hospital, including 538 patients with mild acute pancreatitis (MAP) and 461 patients with MSAP. MSAP patients were divided into four groups according to local complication and transient organ failure (TOF), including single acute peripancreatic fluid collection (APFC) without TOF group (group A), multiple APFC without TOF group (group B), other local complication without TOF group (group C) and TOF group (group D). The baseline data and the severity of AP among the four subgroups were compared. Meanwhile, the severity of disease between group A and MAP patients was also compared. Logistic regression analysis was used to evaluate the risk factors of MSAP. Results: Patients in group D were older than those in group A (P<0.05). There were statistically significant differences in different scoring systems among the four subgroups (P<0.05). The proportions of APACHE Ⅱ≥8, Glasgow≥3 and BISAP≥3 in group D were significantly higher than those in the other three groups (P<0.05). There were significant differences in levels of Ca