Objective To observe the release of inflammation-related factors after angiotensin Ⅱ (Ang Ⅱ ) stimulation in rat tubular epithelial cells (NRK-52E), to analyze whether these effects were mediated by TLR4-MyD88 pathway, and to reveal the novel mechanism of injury by Ang Ⅱ on NRK-52E cells. Methods After synchronization, cells incubated with AngⅡ (10-7 mmol/L) were used as the stimulation group, cells without stimulation were as normal control. To determine the role of TLR4 and the adaptor MyD88, equal number of NRK-52E cells was added with 10-5 mmol/L candesartan or 20 mg/L TLR4 blocking peptide for 1 h and then incubated with Ang Ⅱ (10-7 mmol/L) respectively. RT-PCR was used to analyze TLR4 mRNA and MyD88 mRNA expression. Immunofluorescence and confocal microscopy were used to observe TLR4 protein expression. ELISA was used to detect the concentration of tumor necrosis factor-alpha (TNF-α) and heat shock protein 47(HSP47) in cell supernatant respectively. Results TLR4 and MyD88 were highly expressed in Ang Ⅱ-induced NRK-52E cells (P＜0.01), and the TNF-α and HSP47 levels were also increased markedly compared with control group (P＜0.01). In NRK-52E cells that were pre-incubated with candesartan, TLR4 and MyD88 expression were obviously inhibited,subsequently, HSP47 and TNF-α production decreased remarkably compared with Ang Ⅱ group (P＜0.01). TLR4 blocking peptide had the similar effect in a dose-dependent manner, in which its effect was dependent on inhibiting TLR4-MyD88 expression. Conclusion The mechanism of Ang Ⅱ -induced injury effect on NRK-52E cells is related to the increase of TLR4-MyD88 activity,which is followed by the enhance of TNF-α and HSP47 expression. This process is inhibited by candesartan via modulation of innate immune pathway.

ObjectiveTo investigate the clinical efficacy of fire needling plus filiform needles in treating knee osteoarthritis and its effect on the patients’ quality of life.MethodOne hundred and one patients with knee osteoarthritis were randomly allocated to fire needling, filiform needle and combination groups. They were treated with fire needling, filiform needles and fire needling plus filiform needles, respectively. An assessment was made using the WOMAC Osteoarthritis Index and the 36-item Short Form Health Survey (SF-36) in the 101 patients with knee osteoarthritis before and after treatment.ResultFire needling, filiform needles and fire needling plus filiform needles all had a marked therapeutic effect on knee osteoarthritis. The effect of fire needling plus filiform needles was better than those of fire needling and filiform needles (P<0.05,P<0.01). After four weeks of treatment, SF-36 quality of life item sub-scores increased in all the three groups of patients compared with before treatment (P<0.01); SF-36 role physical and general health scores were higher in the combination group than in the fire needling group (P<0.05, P<0.01); SF-36 physical functioning, role physical, general health, social functioning and mental health scores were higher in the combination group than in the filiform needle group (P<0.01).ConclusionFire needling plus filiform needles can effectively treat knee osteoarthritis and improve the patients’ quality of life. Its effects are superior to those of fire needling and filiform needles.

Objective To observe and compare the clinical efficacy of fire needling versus electroacupuncture in treating knee osteoarthritis of kidney and marrow deficiency type.Methods One hundred and ten patients with knee osteoarthritis of kidney and marrow deficiency type were randomly allocated to two groups, a fire needling group of 56 cases and an electroacupuncture group of 54 cases. The fire needling group received filiform needle acupuncture and fire needle pricking and the electroacupuncture group, electroacupuncture. Treatment was given once every other day, for a total of four weeks as one course. The Visual Analogue Scale (VAS) score and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score were recorded in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups.Results The VAS score and the WOMAC subscores significantly decreased in both groups after treatment compared with those before treatment (P<0.01). There were no statistically significant post-treatment differences in the VAS score and the WOMAC subscores between the two groups (P>0.05). The VAS score and the WOMAC subscores decreased further in both groups at four weeks after treatment compared with those at the end of treatment (P<0.01). In the electroacupuncture group, there were statistically significant differences in the VAS score and the WOMAC pain and stiffness subscores between eight weeks and four weeks after treatment (P<0.01) and in the WOMAC pain, stiffness and joint function subscores between at eight weeks after treatment and at the end of treatment (P<0.01). In the fire needling group, there were statistically significant differences in the VAS score and the WOMAC pain, stiffness and joint function subscores between at eight weeks after treatment and at the end of treatment (P<0.01). At four weeks after treatment, the VAS score and the WOMAC pain, stiffness and joint function subscores were significantly lower in the fire needling group than in the electroacupuncture group (P<0.05,P<0.01). At eight weeks after treatment, the VAS score and the WOMAC pain and stiffness subscores were still significantly lower in the fire needling group than in the electroacupuncture group (P<0.01). The total efficacy rate was 94.6% in the fire needling group and 90.7% in the electroacupuncture group; there was no statistically significant difference between the two groups (P>0.05).Conclusions Both filiform needle acupuncture plus fire needle pricking and electroacupuncture can markedly relieve the pain and stiffness and improve joint function in knee osteoarthritis of kidney and marrow deficiency type. Both have an equivalent short-term therapeutic effect. Filiform needle acupuncture plus fire needle pricking has a better long-term therapeutic effect than electroacupuncture.

To produce recombinant phospholipase A(1) (PLA(1)) by Escherichian coli, the pla gene encoding PLA(1) was amplified from Serratia liquefaciens by PCR and cloned into two vectors pET20-b(+) and pET28-a(+). The two recombinant plasmids were then transformed into E. coli BL21 (DE3) individually to express PLA(1). E. coli BL21(DE3)/pET28a-pla yielded extracellular PLA(1) with an activity of 40.8 U/mL in batch cultivations of shaken flasks by auto-induction, which was accounted for 91% of total enzyme activity. On the basis of primal auto-induction medium, the optimized fermentation medium of PLA(1) contained tryptone 10 g/L, yeast extract 5 g/L, glucose 0.8 g/L, lactose 5 g/L, Na2HPO4 25 mmol/L, KH2PO4 25 mmol/L and 1 mmol/L MgSO4 (final concentration). Glycine (7.5 g/L) was added 6 h after inoculated. After incubated at 37 degrees C for 24 h, extracellular enzyme activity reached 128.7 U/mL.
Cloning, Molecular ; Culture Media ; Escherichia coli ; genetics ; growth & development ; metabolism ; Fermentation ; Lactose ; pharmacology ; Phospholipases A1 ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Serratia liquefaciens ; enzymology

AIM: To observe the effect of Esketamine combined with sufentanil on analgesia and emotion after thoracoscopic radical resection of lung cancer. METHODS: A total of 108 patients undergoing thoracoscopic radical resection of lung cancer were randomly divided into three groups: Esketamine plus sufentanil patient-controlled analgesia group (S group), low-dose sufentanil patient-controlled analgesia group (LC group) and high-dose sufentanil patient-controlled analgesia group (HC group). PCIA was performed after operation. The formula of analgesia pump in group S: Esketamine 1.2 mg/kg, sufentanil 1 μg/kg, LC group analgesic pump formula: sufentanil 1 μg/kg; Formula of analgesic pump in HC group: sufentanil 1.5 μg/kg; Tropisetron 10 mg was added to three groups of analgesia pumps, and normal saline was diluted to 100 mL. Observe the visual analogue scale (VAS) and Ramsay sedation score of resting and exercise pain at 6 h (T

OBJECTIVETo observe the differences in the clinical therapeutic effects on insomnia of spleen and stomach disharmony pattern in comparison of the combination of acupoint catgut-embedding therapy and auricular point pressure with the simple application of auricular point pressure.

METHODSOne hundred and eighty patients of insomnia of spleen and stomach disharmony pattern were randomized into an observation group (93 cases) and a control group (87 cases). In the observation group, the combination of acupoint catgut-embedding therapy and auricular point pressure was adopted. The acupoint catgut-embedding therapy was applied at Zhongwan (CV 12), Anmian (Extra), Xinshu (BL 15), Pishu (BL 20), Weishu (BL 21), Zusanli (ST 36), Yinlingquan (SP 9) and Sanyinjiao (SP 6), once every two weeks. The auricular point pressure was applied to Shenmen (TF), Pizhixia (AT), Xin (CO), Pi (CO), Wei (CO) on both sides alternatively, twice a week. The duration of treatment was 2 months for the two groups. In the control group, the auricular point pressure was simply used with the same points, manipulation and treatment duration as the observation group. After treatment, the clinical therapeutic effects were observed in the patients of the two groups. The Pittsburgh sleep quality index (PSQI) score was compared before and after treatment in the two groups. The recurrence was followed up in 3 months after the end of treatment.

RESULTSIt was observed that 2 cases were dropped out in the observation group, 4 cases in the control group. The dropped out cases were considered to be ineffective. After treatment, the total effective rate was 92.5% (86/93) in the observation group and was 80.5% (70/87) in the control group. The result in the observation group was better than that in the control group (<0.05). After treatment, PSQI scores were all reduced apparently in the two groups as compared with those before treatment (both<0.05). The score after treatment and the different value before and after treatment in the observation group were lower than those in the control group (both<0.05). In 3-month follow-up after treatment, the recurrence rate was 3.2% (3/93) in the observation group, which was lower than 13.8% (12/87) in the control group (<0.05).

CONCLUSIONIn the treatment of insomnia of spleen and stomach disharmony pattern, the acupoint catgut-embedding therapy combined with auricular point pressure are superior to the simple application of auricular point pressure in the clinical therapeutic effects, improving sleep quality and reducing the recurrence rate.