[Clinical analysis of 555 outpatients with hand, foot and mouth diseases caused by different enteroviruses].

Objective: To study the clinical characteristics of outpatients with hand, foot and mouth disease (HFMD) caused by different serotypes of enteroviruses. Methods: This was a prospective study. From February 2017 to March 2018, 563 outpatients with HFMD were enrolled by systematic sampling in the Department of Infectious Diseases, Henan Children's Hospital. Throat swabs were collected to determine the serotypes via PCR. Demographic, clinical, and laboratory data were collected by standard questionnaire. All cases were followed up twice at 2 and 9 weeks after the initial outpatient visit through telephone interview. A total of 563 cases were enrolled and 555 (98.6%) cases were positive for human enteroviruses, including 338 (60.9%) males. Analyses were stratified by enterovirus serotypes, Chi square test or Fisher's exact test, Rank sum test was used for comparison among different groups. Results: The age of 555 cases was 24.2 (16.4, 41.3) months. Among them 44.0% (224 cases) were identified as coxsackievirus (CV)-A6, while 189 cases, 35 cases, 14 cases and 73 cases were identified as CV-A16, enterovirus (EV)-A71, CV-A10 and other serotypes, respectively. Fever (≥37.5 ℃) was present in 51.4% (285/555) of laboratory confirmed cases. The proportions of fever in cases of CV-A6 (68.9%(168/244)) and CV-A10 (12/14) were significantly higher than those in cases of CV-A16 (31.7%(60/189),χ(2)=57.344,14.313,both P=0.000), other serotypes (43.8%(32/73),χ(2)=15.101 and 8.242, P=0.000 and 0.004) and EV-A71 (37.1%(13/35), χ(2)=13.506 and 9.441, P=0.000 and 0.002) respectively. There was no significant difference between CV-A6 and CV-A10 in presentation of fever (χ(2)=1.785, P=0.182). There were 359 cases (64.7%) with eruptions in mouth, hands, feet and buttocks. Cases infected with EV-A71 had the highest proportions (74.3%(26/35)) of rash emerging simultaneously in mouth, hands, feet, and buttocks. The proportion in cases of CV-A16, CV-A6, CVA10 and other serotype were 73.5% (139/189), 61.9% (151/244), 7/14 and 49.3% (36/73), respectively. The proportion of rash on other parts of body, such as face, limbs or torso in cases infected with CV-A6 (16.8% (41/244)) was the higherest and the proportion in cases of CV-A16, EV-A71, CV-A10 or other serotypes were 8.5% (16/189) , 5.7% (2/35) , 1/14, 6.8% (5/73) , respectively. None of these cases developed serious complications. Desquamation occurred in 45.5% (179/393) cases 7.5 (5.0, 9.0) days after disease onset and 13.5% (53/393) cases showed onychomadesis 31.0 (18.0, 33.5) days after disease onset. The proportion of desquamation and onychomadesis associated with CV-A6 (64.2% (95/148) and 31.8% (47/148)) was significantly higher than CV-A16 (31.8% (49/154) and 1.3% (2/154), χ(2)=33.601 and 52.482, both P=0.000) and other serotypes (38.0%(19/50) and 6.0%(3/50),χ(2)=10.236 and 12.988, P=0.001 and 0.000). Desquamation appeared more in cases of CV-A6 than in cases of CV-A10 (2/11,χ(2)=9.386, P=0.002), with the proportion of onychomadesis higher in CV-A6 than in EV-A71 (3.3% (1/30),χ(2)=11.088, P=0.001). Conclusion: Clinical manifestation such as fever, rash emerging parts, desquamation and onychomadesis are different among outpatient HFMD cases infected with CV-A16, CV-A6, EV-A71, CV-A10 and other enteroviruses.

Protosappanin a, an immunosuppressive constituent from a Chinese herb, prolongs graft survival and attenuates acute rejection in rat heart allografts.

OBJECTIVE: Caesalpinia sappan L., which has been used in oriental medicine as an analgesic and antiinflammatory agent, has shown immunosuppressive activity on acute rejection on rat heart allografts. The present study was designed to investigate the potency of protosappanin A, one major ingredient of C. sappan L., in rat heart transplantation. MATERIALS AND METHODS: Protosappanin A isolated from an ethanol extract of C. sappan L. was identified by wave spectrum. All groups consisted of Wistar donors into Sprague-Dawley (SD) recipients, including large (25 mg/kg) or small (5 mg/kg) doses of protosappanin A plus cyclosporine (10 mg/kg). A control group was used. Animals were given the drugs on postoperative day 2. We observed graft survival and pathologic conditions of the hearts. Blood samples were obtained on postoperative day 7 to measure the CD4+/CD8+ T-cell ratio. Graft perforin and granzyme B mRNA expression levels were examined with reverse transcription-polymerase chain reaction (RT-PCR) methods. RESULTS: Protosappanin A or cyclosporine significantly prolonged heart allograft survival (P < .01), alleviated myocardial pathologic damages (P < .01), decreased the CD4+/CD8+ ratio (P < .05), and inhibited perforin and granzyme B mRNA expressions in the graft (P < .05). CONCLUSIONS: These findings demonstrated that protosappanin A prolonged heart allograft survival by significantly attenuating acute rejection.

The catalytic role of subunit IV of the cytochrome b6-f complex from spinach chloroplast.

The catalytic role of subunit IV, the Mr 17,000 protein, in the chloroplast cytochrome b6-f complex was established through trypsinolysis of the complex under controlled conditions. When purified chloroplast cytochrome b6-f complex, 1 mg/ml, in 50 mM Tris-succinate buffer (pH 7.0) containing 1% sodium cholate and 10% glycerol is treated with 80 micrograms of trypsin at room temperature for various lengths of time, the activity of the cytochrome b6-f complex decreases as the incubation time increases. A maximal inactivation of 80% is reached at 7 min of incubation. The trypsin inactivation is accompanied by the destruction of the proton translocation activity of the complex. No alteration of absorption and EPR spectral properties was observed in the trypsin-inactivated complex. Subunit IV is the only subunit in the cytochrome b6-f complex that is digested by trypsin, and the degree of digestion correlates with the decrease of electron transfer activity. The binding of azido-Q to subunit IV of the complex decreases as the extent of inactivation of the cytochrome b6-f complex by trypsin increases. The residue molecular mass of trypsin cleaved subunit IV is about 14 kDa, suggesting that the cleavage site is at lysine 119 or arginine 125 or 126. When the thylakoid membrane was assayed for cytochrome b6-f complex activity, very little activity was observed; and the activity was not sensitive to trypsinolysis. Upon sonication, activity and sensitivity to trypsinolysis was greatly increased, suggesting that subunit IV protrudes from the lumen side of the membrane.

Crystallization of mitochondrial ubiquinol-cytochrome c reductase.

Ubiquinol-cytochrome c reductase of beef heart mitochondria was crystallized in the presence of decanoyl-N-methylglucamide, heptanetriol, and sodium chloride with poly(ethylene glycol) as precipitant. The largest crystal has dimensions of 4 x 2 x 1 mm. The crystalline enzyme is composed of 10 subunits. It contains 2.5 nmol of ubiquinone, 8.4 nmol of cytochrome b, 4.2 nmol of cytochrome c1, 4.2 nmol of iron-sulfur cluster, and 140 nmol of phospholipid per milligram of protein. Of the last, 36% is with diphosphatidylglycerol. The crystals are very stable in the cold and show full enzymatic activity when redissolved in aqueous solution. Absorption spectra of the redissolved crystals show a Soret to UV ratio of 0.88 and 1.01 in the oxidized and the reduced forms, respectively.