Optimization of production process of Aeromonas veronii CA07 strain inactivated vaccine / 中国生物制品学杂志

@#Abstract：Objective To optimize the production process of inactivated vaccine of Aeromonas veronii（AV）CA07 strain. Methods The fermentation culture process of AV CA07 strain liquid was determined through the optimization of the culture time（2～16 h），medium（optimized fermentation medium，LB medium and NB medium）and fermentation conditions（in⁃ oculation amount of 1%，5%，10% and 15%；ventilation rate of 2，4，6 and 8 L／min and fermentation time of 6，8，10 and 12 h）. The optimal inactivation process was determined through the comparison of the final concentration of formalde⁃ hyde solution（0. 10%，0. 20%，0. 30% and 0. 40%），inactivation temperature（28 and 37 ℃）and inactivation time（24， 48 and 72 h）. The large⁃scale production process of inactivated vaccine of AV CA07 strain in 500 L fermentor was estab⁃ lished and the prepared vaccines were tested for safety and immunogenicity. Results The optimal inoculation amount of AV CA07 strain was 5%，ventilation rate was 4 L／min and culture time was 10 ~ 12 h. The optimal inactivation condition was adding formaldehyde solution with final concentration of 0. 30% incubating at 37 ℃ for 24 h. The number of viable bacteria in the fermentation broth of AV CA07 strain prepared in 500 L fermentor was more than 8 × 109 CFU／mL. All crucian carps immunized with the inactivated vaccine by abdomen survived. After challenge，the relative immune protection rate was more than 90%. Conclusion AV CA07 strain inactivated vaccine prepared by optimized production process showed good safety and immunogenicity.

Subcutaneous tissue-sparing dorsal slit with new marking technique: A novel circumcision method.

Circumcision is a very common surgical procedure that is performed for medical and traditional purposes in the world. However, many technical of circumcision is needed to improve. Thus, this study introduced a novel method of circumcision that is a refined version of the sleeve technique, termed subcutaneous tissue sparing dorsal slit with new marking, and evaluated the safety and efficacy of this novel method of circumcision.The randomized clinical trial included 93 adult patients with redundant foreskin or phimosis treated from May 2015 to March 2017. Patients were randomly divided into the novel circumcision method (n = 45) or conventional dissection (n = 48). The groups were compared regarding rates of intraoperative hemorrhage, operative time, pain, healing, satisfaction with penis appearance, and relevant adverse events.No patient suffered any obvious complication. Compared with the patients given conventional dissection, the patients who underwent the new surgical device experienced significantly less wound healing time, scar width, and recovery time (P <.05). The new method resulted in greater intraoperative bleeding volume and surgical time (P <.05). The rate of satisfaction with appearance of the penis was significantly higher in the group treated with the novel technique. In addition, the cost of surgery of these 2 methods was similar.Based on the above research, we found that subcutaneous tissue-sparing dorsal slit with new marking technique was an effective and safe procedure for circumcision, and deserved further application in clinical practice.

20-Hydroxyeicosatetraenoic acid contributes to the inhibition of K+ channel activity and vasoconstrictor response to angiotensin II in rat renal microvessels.

The present study examined whether 20-hydroxyeicosatetraenoic acid (HETE) contributes to the vasoconstrictor effect of angiotensin II (ANG II) in renal microvessels by preventing activation of the large conductance Ca(2+)-activated K(+) channel (KCa) in vascular smooth muscle (VSM) cells. ANG II increased the production of 20-HETE in rat renal microvessels. This response was attenuated by the 20-HETE synthesis inhibitors, 17-ODYA and HET0016, a phospholipase A2 inhibitor AACOF3, and the AT1 receptor blocker, Losartan, but not by the AT2 receptor blocker, PD123319. ANG II (10(-11) to 10(-6) M) dose-dependently decreased the diameter of renal microvessels by 41 ± 5%. This effect was blocked by 17-ODYA. ANG II (10(-7) M) did not alter KCa channel activity recorded from cell-attached patches on renal VSM cells under control conditions. However, it did reduce the NPo of the KCa channel by 93.4 ± 3.1% after the channels were activated by increasing intracellular calcium levels with ionomycin. The inhibitory effect of ANG II on KCa channel activity in the presence of ionomycin was attenuated by 17-ODYA, AACOF3, and the phospholipase C (PLC) inhibitor U-73122. ANG II induced a peak followed by a steady-state increase in intracellular calcium concentration in renal VSM cells. 17-ODYA (10(-5) M) had no effect on the peak response, but it blocked the steady-state increase. These results indicate that ANG II stimulates the formation of 20-HETE in rat renal microvessels via the AT1 receptor activation and that 20-HETE contributes to the vasoconstrictor response to ANG II by blocking activation of KCa channel and facilitating calcium entry.

[The clinical analysis of mercury poisoning in 92 cases].

OBJECTIVE: To summarize the clinical features of mercury poisoning diagnosed by blood and urine tests for improving the diagnosis and treatment of the disease. METHODS: Poisoning causes, clinical manifestations, diagnosis, treatment and prognosis were retrospectively reviewed in 92 in-patients with mercury poisoning in our hospital from January 2000 to April 2010. RESULTS: Of the 92 patients, 37 were male and 55 were female with an average age of 33.1 (2 - 65) years old. The mercury poisoning was caused by occupational exposure and non-occupational exposure, such as iatrogenic exposure, life exposure and wrong intake or suicidal intake of mercury-containing substances, mainly through respiratory tract, digestive tract and skin absorption. The most common clinical symptoms were as the followings: nervous system symptom, such as memory loss in 50 cases (54.3%), fatigue in 34 (37.0%), numb limb in 25 (27.2%), dizziness and headache in 22 (23.9%), cacesthesia in 20 (21.7%), fine tremor (finger tip, tongue tip, eyelids) in 15 (16.3%), insomnia and more dreams in 12 (13.0%); gastrointestinal symptoms: nausea in 16 (17.4%), abdominal pain in 14 (15.2%), stomatitis in 5 (5.4%); joint and muscle symptoms: muscle pain in 16 (17.4%), joint pain in 5 (5.4%); cardiovascular system: chest tightness, heart palpitations in 6 (6.5%); urinary system: edema in 9 (9.8%); other system: hidrosis in 20 (21.7%). After the treatment with sodium dimercaptopropane sulfonate (DMPS), the symptoms were gradually alleviated. Their gastrointestinal, cardiovascular symptoms were alleviated within 2 weeks; neurological symptoms were alleviated within 3 months; kidney damage showed a slower recovery and could be completely alleviated within 6 months. CONCLUSIONS: Because of its diverse clinical symptoms, the mercury poisoning was easy to misdiagnosis and missed diagnosis; therefore the awareness of the disease should be further enhanced. Leaving from the poisoning environment timely and giving appropriate treatment with DMPS will lead to a satisfactory prognosis.

Morton lentil extract attenuated angiotensin II-induced cardiomyocyte hypertrophy via inhibition of intracellular reactive oxygen species levels in vitro.

The objective was to investigate whether a lentil (Morton) extract had any protective effect on cardiac hypertrophy, which is one of the most significant sequelae of cardiovascular diseases. High phenolic compounds (43.4 mg of GAE/g), including thirteen phenolic acid and two flavonoids, were detected in the acetone/water/acetic acid lentil extract. The extract showed strong antioxidant ability (105 µmol of TE/g). The effect of lentil extract on angiotensin (Ang) II-induced cardiac hypertrophy was examined. Results showed that pretreatment with lentil extract (25, 50, 100 µg/mL) significantly attenuated Ang II (0.1 µM)-induced hypertrophy by 18, 28, and 36% in rat cardiomycytes, respectively; lentil extract (12.5, 25, 50 µg/mL) attenuated Ang II (0.1 µM)-induced hypertrophy by 9, 17, and 25% in human cardiomycytes, respectively. Intracellular reactive oxygen species (ROS) levels were enhanced by Ang II treatment, and this stimulatory action was significantly attenuated (33% inhibition) by lentil extract (100 µg/mL) in rat cardiomyocytes and attenuated by 22% by 50 µg/mL lentil extract in human cardiomyocytes. In conclusion, Morton lentil extracts attenuated Ang II-induced rat and human cardiomyocytes hypertrophy via decreasing intracellular ROS levels.

Effect of angiotensin II type 1 receptor on delayed rectifier potassium current in catecholaminergic CATH.a cells.

AIM: To study the modulatory effects of angiotensin II (Ang II) on the delayed rectifier potassium (Kv) current (IKv) and its underlying intracellular mechanism in the catecholaminergic system of rats. METHODS: AT1 and AT2 receptors of the differentiated and undifferentiated CATH.a cells were determined by radioligands binding assay. The IKv was recorded with the whole cell patch-clamp configuration in voltage clamp mode on CATH.a cells. RESULTS: The Ang II receptor proteins including AT1 and AT2 receptors were expressed in CATH.a cells, and the number of the former was significantly more than the latter (P<0.01). The IKv of CATH.a cells was reduced by superfusion with the Ang II (100 nmol/L) (P<0.05) in the presence of the AT2 receptor antagonist PD123319, but was not affected by only superfusion with PD123319. The effect of Ang II on IKv in CATH.a cells was completely inhibited by addition of AT1 receptor antagonist losartan. Superfusion with Ang II (100 nmol/L) plus U73122, an inhibitor of phospholipase C (PLC) in the presence of PD123319 had no effect on the IKv [(20.2+/-2.8) pA/pF]. Ang II-induced reduction of IKv was attenuated (P<0.05) but not abolished by PKC inhibitor calphostin C (Cal) and selective CaMK II inhibitor KN-93 (10 micromol/L) respectively. However, IKv reduction was completely abolished by superfusion with both Cal and KN-93. CONCLUSION: The inhibition of Kv currents in CATH.a cells by Ang II is mediated by AT1 receptor, and the PLC, PKC and CaMK II may be involved in signal transduction of AT1 receptor. The differentiated CATH.a cell is a useful cell model to study Ang II receptor-mediated functional modulation of catecholaminergic system.

20-hydroxyeicosatetraenoic acid (20-HETE): structural determinants for renal vasoconstriction.

The effects of natural and synthetic eicosanoids on the diameter of rat interlobular arteries studied in vitro were compared to that of the potent, endogenous vasoconstrictor 20-HETE. Vasoconstrictor activity was optimum for chain lengths of 20-22 carbons with at least one olefin or epoxide between located between C(13)-C(15) and an oxygen substituent at C(20)-C(22). The presence of delta (Zou et al. Am. J. Physiol. 1996, 270, R228; Gebremedhin, D. et al. Am. J. Physiol. 1998, 507, 771)-, delta (Carroll et al. Am. J. Physiol. 1996, 271, R863; Vazquez et al. Life Sci. 1995, 56, 1455)-, or delta (Imig et al. Hypertension 2000, 35, 307; Lopez et al. Amer. J. Physiol. 2001, 281, F420)-olefins had no influence on the vasoconstrictor response whereas the introduction of a C(7)-thiomethylene enhanced potency. A sulfonamide or alcohol, but not a lactone, could replace the C(1)-carboxylate. These data were used to construct a putative binding domain map of the 20-HETE receptor consisting of: (i) a comparatively open, hydrophilic binding site accommodating the C(1)-functionality; (ii) a hydrophobic trough spanning the olefins; (iii) a shallow pocket containing a critical pi-pi binding site in the vicinity of the pi (Ito et al. Am. J. Physiol. 1998, 274, F395; Quigley, R.; Baum, M.; Reddy, K. M.; Griener, J. C.; Falck, J. R. Am. J. Physiol. 2000, 278, F949)-olefin; and (iv) an oxyphilic binding site proximate to the omega-terminus.

Mechanism of cGMP contribution to the vasodilator response to NO in rat middle cerebral arteries.

This study examined the mechanism by which cGMP contributes to the vasodilator response to nitric oxide (NO) in rat middle cerebral arteries (MCA). Administration of a NO donor, diethylaminodiazen-1-ium-1,2-dioate (DEA-NONOate), or 8-bromo-cGMP (8-BrcGMP) increased the diameter of serotonin-preconstricted MCA by 79 +/- 3%. The response to DEA-NONOate, but not 8-BrcGMP, was attenuated by iberiotoxin (10(-7) M) or a 80 mM high-K(+) media, suggesting that activation of K(+) channels contributes to the vasodilator response to NO but not 8-BrcGMP. The effects of NO and cGMP on the vasoconstrictor response to Ca(2+) were also studied in MCA that were permeabilized with alpha-toxin and ionomycin. Elevations in bath Ca(2+) from 10(-8) to 10(-5) M decreased the diameter of permeabilized MCA by 76 +/- 5%. DEA-NONOate (10(-6) M) and 8-BrcGMP (10(-4) M) blunted this response by 60%. Inhibition of guanylyl cyclase with 1H-[1,2,4]oxadiazole[4,3-a] quinoxalin-1-one (10(-5) M) blocked the inhibitory effect of the NO donor, but not 8-BrcGMP, on Ca(2+)-induced vasoconstriction. 8-BrcGMP (10(-4) M) had no effect on intracellular Ca(2+) concentration ([Ca(2+)](i)) in control, serotonin-stimulated, or alpha-toxin- and ionomycin-permeabilized vascular smooth muscle cells isolated from the MCA. These results indicate that the vasodilator response to NO in rat MCA is mediated by activation of Ca(2+)-activated K(+) channels via a cGMP-independent pathway and that cGMP also contributes to the vasodilator response to NO by decreasing the contractile response to elevations in [Ca(2+)](i).