Pharmaceutical water is the most widely used raw material and solvent in the process of pharmaceutical products, which is widely used in the cleaning and disinfecting process of appliances, equipment, facilities and systems, and is involved in the whole process of pharmaceutical products. Pharmaceutical water is one of the main sources of microbial contamination of pharmaceutical products, so the microbial safety of pharmaceutical water is crucial for pharmaceutical quality. This paper mainly introduces the characterization of the pharmaceutical water system, and conducts biofilm analysis. Due to the characteristics of oligotrophic and slow-growing of microorganisms in the pharmaceutical water system, this paper also further discusses the microbiological testing and culture methods of pharmaceutical water, so as to provide reference for the establishment of a scientific and reasonable microbiological testing system, as well as provide an important theoretical basis and new ideas for further research on hard-to-culture microorganisms.

Objective:To investigate the effect of plasma exosome miR-18a-3p on of apoptosis and cell cycle cumulus cells (CCs) in polycystic ovary syndrome (PCOS) via targeting CITED2.Methods:qRT-PCR assay was used to detect the expression of miR-18a-3p in plasma, plasma-derived exosome and CCs of patients no matter with PCOS and non PCOS. After being transfected with miR-18a-3p mimic, the exosome was co-cultured with CCs, the cell cycle distribution and apoptosis of CCs were detected by flow cytometry assays. Gene ontology (GO) analysis was performed and CITED2 was included in follow up experiments. pcDNA3.1-CITED was transfected into CCs and then co-cultured with exosome to explore the co-effect of miR-18a-3p and CITED2 on the cell cycle and apoptosis of CCs.Results:The plasma-derived exosome isolated from CCs with PCOS were identified successfully, and the expression of miR-18a-3p was significantly decreased in plasma, exosome and CCs in PCOS, compared with that in non-PCOS (all P<0.05) . CITED2 could be regulated as a target of miR-18a-3p in CCs. Compared with NC group, overexpression of miR-18a-3p could significantly decrease proportion of CCs cells in G0/G1 phase and inhibit their apoptosis in PCOS patients (all P<0.05) . The effect of over-expressing miR-18a-3p could be partially reversed by up-regulating CITED2 (all P<0.05) . Conclusions:Plasma exosomal miR-18a-3p has the effect to induce the S phase of CCs cells, subsequently inhibit apoptosis, then restrain the progression of PCOS. Plasma exosomal miR-18a-3p is expected to play a paramount role in the target therapy of PCOS.

Objective:To investigate the role of WWC2-AS1/miR-382-5p/FZD3 in granulosa cell (GCs) of polycystic ovary syndrome (polycystic ovarian syndrome, PCOS) patients and its molecular mechanism.Methods:Bioinformatics tools were used to predict the molecular mechanism of PCOS. The expressions of WWC2-AS1, miR-382-5p and FZD3 in serum and GCs of patients with PCOS and healthy controls were detected by qRT-PCR. The effects of WWC2-AS1/miR-382-5p/FZD3 on the proliferation and apoptosis of GCs were observed by CCK-8 and flow cytometry. The interaction between WWC2-AS1 and miR-382-5p, miR-382-5p and FZD3 was verified by double luciferase report experiment.Results:Compared with the control group, the expression of WWC2-AS1 and FZD3 in serum and GCs of PCOS patients was significantly up-regulated, while the expression of miR-382-5p was down-regulated. Silencing WWC2-AS1 could significantly promote the proliferation of GCs in PCOS and inhibit the apoptosis of GCs (all P<0.05) . There is a WWC2-AS1/miR-382-5p/FZD3 interaction network in PCOS, and miR-382-5p inhibitor or overexpressed FZD3 can partially reverse the regulatory effect of silent WWC2-AS1 on GCs in PCOS. Conclusion:This study shows that WWC2-AS1 regulates miR-382-5p and up-regulates FZD3, which promotes the proliferation of GCs and inhibits apoptosis in the progression of PCOS. WWC2-AS1/miR-382-5p/FZD3 may be an effective molecular target for the treatment of PCOS.

Objective To investigate the effects of inhibiting TIM-4 function in Kupffer cells (KCs) on liver graft rejection in mice and explore the underlying mechanism. Methods Mouse models of orthotopic liver transplantation were treated with a control mAb group and TIM-4 mAb. The activated KCs were assayed with immunohistochemistry after operation. The expression of TIM-4 in KCs were assayed with Western blotting and RT-PCR and the levels of AST, ALT, TBIL, TNF-α, IFN-γand CCL2 were assayed detected. The expression of TIM-4 in KCs was observed with laser confocal microscopy. HE staining was used to observe the microstructure of the liver tissues, and the number of CD25 +Foxp3 +T cells was determined using with flow cytometry; the proteins levels of p-P65and p-P38 were assayed with Western blotting. The donor mice were treated with clodronate liposomes to destroy the KCs in the liver before transplantation, and the liver grafts were examined for graft rejection. Results The number of activated KCs in the liver graft increased progressively over time. Compared with the sham-operated group, the liver graft showed significantly increased TIM-4 protein and mRNA levels at 1, 3, and 7 days after transplantation (P<0.05) and increased levels of AST, ALT, TBIL, TNF-α, IFN-γ and CCL2 at 7 days (P<0.05). The graft in TIM-4 mAb group showed mild pathological changes with a mean RAI score of 2.67 ± 0.75, which was significantly lower than that in control mAb group (P<0.05). The mean survival time of the recipient mice was 53.8±6.4 days in TIM-4 mAb group, significantly longer than that in the control mAB group (14.5±2.9 days, P<0.05). Donor treatment with clodronate liposomes resulted in comparable RAI scores in TIM-4 mAb and control mAb groups (8.01±0.64 vs 7.93±0.56, P>0.05). Theprotein levels of p-P65 and p-P38 in TIM-4 mAb group were significantly lower than those in control mAb group (P<0.05), and CD25+Foxp3+T cells in the liver graft increased significantly in TIM-4 mAb group. Conclusion Inhibition of TIM-4 function in KCs reduces the production of inflammatory factors after liver transplantation possibly by inhibiting the NF-κB and MAPK signaling pathways and promoting the proliferation of Foxp3+Treg cells to induce allograft tolerance.

Objective To investigate the effects of inhibiting TIM-4 function in Kupffer cells (KCs) on liver graft rejection in mice and explore the underlying mechanism. Methods Mouse models of orthotopic liver transplantation were treated with a control mAb group and TIM-4 mAb. The activated KCs were assayed with immunohistochemistry after operation. The expression of TIM-4 in KCs were assayed with Western blotting and RT-PCR and the levels of AST, ALT, TBIL, TNF-α, IFN-γand CCL2 were assayed detected. The expression of TIM-4 in KCs was observed with laser confocal microscopy. HE staining was used to observe the microstructure of the liver tissues, and the number of CD25 +Foxp3 +T cells was determined using with flow cytometry; the proteins levels of p-P65and p-P38 were assayed with Western blotting. The donor mice were treated with clodronate liposomes to destroy the KCs in the liver before transplantation, and the liver grafts were examined for graft rejection. Results The number of activated KCs in the liver graft increased progressively over time. Compared with the sham-operated group, the liver graft showed significantly increased TIM-4 protein and mRNA levels at 1, 3, and 7 days after transplantation (P<0.05) and increased levels of AST, ALT, TBIL, TNF-α, IFN-γ and CCL2 at 7 days (P<0.05). The graft in TIM-4 mAb group showed mild pathological changes with a mean RAI score of 2.67 ± 0.75, which was significantly lower than that in control mAb group (P<0.05). The mean survival time of the recipient mice was 53.8±6.4 days in TIM-4 mAb group, significantly longer than that in the control mAB group (14.5±2.9 days, P<0.05). Donor treatment with clodronate liposomes resulted in comparable RAI scores in TIM-4 mAb and control mAb groups (8.01±0.64 vs 7.93±0.56, P>0.05). Theprotein levels of p-P65 and p-P38 in TIM-4 mAb group were significantly lower than those in control mAb group (P<0.05), and CD25+Foxp3+T cells in the liver graft increased significantly in TIM-4 mAb group. Conclusion Inhibition of TIM-4 function in KCs reduces the production of inflammatory factors after liver transplantation possibly by inhibiting the NF-κB and MAPK signaling pathways and promoting the proliferation of Foxp3+Treg cells to induce allograft tolerance.

microRNAs combined with specific mRNAs are 19-25 nucleotide-long small-molecule RNA that mediate sequence-dependent post-transcriptional gene expression.Accumulating evidences indicate that microRNAs target critical signal transduction molecules of immune system,and involve in regulation of immune tolerance.Recently,microRNAs have been a potential biomarker,and are widely useded in diagnosis and prognosis of cancer,infectious disease,autoimmune disease,and transplantation.If we can further identify regulatory mechanism of microRNAs and their target genes,which makes possible the successful induction of immune tolerance and exert a huge push on organ transplantation.

Objective To investigate the relationship between inflammation and blood coagulation function in the patients with acute exacerbation of chronic cor pulmonale (AECCP) and discuss the potential mechanism and influence on the patients. Methods The present study was based on 30 healthy controls and 141 cases of AECCP in our hospital from January 2011 to June 2014.Levels of white blood cell (WBC), neutrophil (NEUT), high-sensitivity C-reactive protein (hs-CRP, Complement 3 (C3), prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and thrombin time (TT) in the patients were determined. Results Compared with the healthy controls, the patients had higher levels of WBC, NEUT, hs-CRP, PT, APTT, FIB, TT (all P < 0.001) and lower level of C3 (P < 0.001). Significant positive correlations were found between the levels of WBC, NEUT and FIB (r = 0.196 and r = 0.199, both P < 0.05); hs-CRP and APTT, FIB(r = 0.234, P < 0.01 and r = 0.466, P < 0.001); C3 and FIB(r = 0.466, P < 0.001), and significant negative correlations were observed between the levels of C3 and PT, APTT, TT (r=-0.258, P<0.01;r=-0.279, P < 0.01 and r = -0.168, P < 0.05, respectively). Compared with the survival patients, the cases of death had higher levels of WBC and NEUT (both P < 0.01). The area under receiver operating characteristic curve of WBC and NEUT, predicting the prognosis, was 0.666 (95% CI 0.552, 0.780; P < 0.01) and 0.695 (95% CI 0.558, 0.801; P = 0.001) respectively. Conclusions Inflammation and blood coagulation function disorder usually coexist in the patients with AECCP, and are closely associated with the severity. Levels of both WBC and NEUT can be used as prognosis predictors for the patients.

Objective To study the transfection effects of nuclear factor-KappaB(NF-κB)decoy oligodeoxynucleotides(ODN) to Kupffer cells (KCs) mediated by lipofectamine,and investigate it's suppression effects on KCs activation. Methods Twenty-four Wistar rats were divided into three groups (n=8).(1)Control group,in which the normal KCs were isolated.(2)LPS group,in which 1 ms/L LPs was added to the culture system.(3)NF-κB decoy ODN group,in which KCs were transduced with NF-κB decoy ODN (4μg×105KCs)prior to LPS stimulation.The transfection efficiency Was assayed,and the phagocytosis function,NF-κB(P65) translocation,CD40 mRNA expression of KCs were also detected respectively. Results Kupffer cells were obviously activated after LPS stimulation.the phagocytosis function was reinforced.the activity of NF-κB transloeated from cytoplasm into nucleus was obviosly increaced.The co-stimulatory molecules expression(CD40 mRNA)significantly increased compared with control group(t=4.01,P<0.01).NF-κB decoy oligodeoxynucleotides can efficiently transfected into KCs mediated by lipofectamine,which can obviously suppress KCs activation,and downregulate the expression of downstream gene(compared with LPS group,t=4.89,P<0.01). Condusion NF-κB decoy ODN can efficiently transfect into KCs and inhibit it's activation.

Objective:To analyze the nosazontology of pharyngeal paraesthesia and investigate the treatment. Method: Two hundred and twelve misdiagnosed pharyngeal paraesthesia patients were investigated by history inquiry,routine examination, 24-hour esophageal pH monitoring, barium X-ray of the oesophagus, anxietas-athy-mia private measuring scale, coefficient of variation of the R-R(CVR-R), bioavailable testosterone detection(Bio-T), erection experiment and questionnaire about man climacteric syndrome. The concomitant symptoms and positions of pharyngeal paresthesia were also studied. We adopted individuallied sequential multi-therapy for every patient according to the cause of disease. Result:The cause of disease within 212 cases of pharyngeal paraesthesia included 62 psychictrauma,32 endocrine system disease,106 upper gastrointestinol disease, circulatory disease,9 circulatory disease,3 idiopathic. With individualized treatment, 110 cases had fully recovered, 63 cases excellence and 31 cases utility,and the efficiency rate was 96.23%. Conclusion:Pharyngeal paraesthesia can be caused by several factors. Thorough examination and comprehensive analysis should be applied to those incurable patient who has been treated for a long time. Short course of treatment and irrational drug use are the main causes of short term recurrence and unsatisfactory curative effect.

OBJECTIVE: To analyze the nosazontology of pharyngeal paraesthesia and investigate the treatment. METHOD: Two hundred and twelve misdiagnosed pharyngeal paraesthesia patients were investigated by history inquiry, routine examination, 24-hour esophageal pH monitoring, barium X-ray of the oesophagus, anxieties-athymic private measuring scale, coefficient of variation of the R-R (CVR-R), bioavailable testosterone detection (Bio-T), erection experiment and questionnaire about man climacteric syndrome. The concomitant symptoms and positions of pharyngeal paresthesia were also studied. We adopted individuallized sequential multi-therapy for every patient according to the cause of disease. RESULT: The cause of disease within 212 cases of pharyngeal paraesthesia included 62 psychic trauma, 32 endocrine system disease, 106 upper gastrointestinal disease, circulatory disease, 9 circulatory disease, 3 idiopathic. With individualized treatment, 110 cases had fully recovered, 63 cases excellence and 31 cases utility, and the efficiency rate was 96.23%. CONCLUSION Pharyngeal paraesthesia can be caused by several factors. Thorough examination and comprehensive analysis should be applied to those incurable patient who has been treated for a long time. Short course of treatment and irrational drug use are the main causes of short-term recurrence and unsatisfactory curative effect.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Paresthesia ; diagnosis ; etiology ; therapy ; Pharyngeal Diseases ; diagnosis ; etiology ; therapy ; Pharynx ; pathology ; Young Adult