Adenosine Triphosphate ; metabolism ; Animals ; Animals, Newborn ; Cantharidin ; pharmacokinetics ; Cell Hypoxia ; drug effects ; Enzyme Inhibitors ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Flow Cytometry ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Swine

The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.

Aim To observe the expression of mesen-cephalic astrocyte-derived neurotrophic factor(MANF) in synovial membrane and serum of rats with adjuvant arthritis (AA) and to analyse the relationship between MANF expression and arthritis. Methods AA models were prepared by injecting Freund complete adjuvant (FCA) into SD rats. The swelling of the secondary joint was measured by foot volume measurement. The severity of AA was recorded by arthritis index (AI). Synovial pathological changes were observed by HE staining. The protein and mRNA levels of MANF,BiP and CHOP extracted from synovial tissues in different periods of AA rats were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The levels of MANF, C-reactive protein (CRP), interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) in serum were detected by enzyme-linked immunosorbent assay (ELISA) and then the relationship between MANF level and inflam-matory factors were explored. Results AA rat model was established successfully. The expression of BiP significantly increased in synovial tissue on d 2 after CFA injection,and decreased until d 28. The expres-sion of MANF slightly increased on d 2,then remained stable,and significantly increased on d 14, and then decreased gradually. The expression of CHOP kept to rise slowly at a low level. The level of MANF in serum markedly increased on d 14,then gradually decreased, but it was still higher than the normal level on d 28. The level of CRP exhibited similar trend with MANF. Correlation analysis showed that MANF had a negative correlation with arthritis symptoms, IL-1β and TNF-α in the secondary inflammatory period of AA rats. Con-clusions Arthritis induces the expression and secre-tion of MANF,and the level of MANF is closely relat-ed to the progression and severity of arthritis.

·AIM: To observe the therapeutic effect of visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia.·METHODS:Totally 73 children ( 135 eyes ) with myopic amblyopia were divided into control group ( 36 cases, 67 eyes) and treatment group (37 cases, 68 eyes) according to random number table. The control group were treated with traditional spectaculars and grating covering combined with fine eyesight training;the treatment group were treated with visual perception training combined with total nutrient meal sequential therapy. The visual acuity, diopter and average diopter of two groups were compared before and after treatment at 3, 6mo and 1a. The curative effect of two groups of children was compared after 1a treatment. And the adverse reactions were recorded in two groups during the treatment period. The recurrence rate of amblyopia in 1a follow-up was compared between two groups.·RESULTS: The difference of visual acuity between two groups was not significant at 3mo (P>0. 05). The visual acuity of the treatment group was significantly higher than that of the control group at 6mo and 1a (P<0. 05). There was no significant difference in diopter between the two groups after 3, 6mo and 1a (P>0. 05), but the average annual refractive changes in the treatment group were significantly lower than that in the control group ( P<0. 05). The basic cure rate and total effective rate of the treatment group were significantly higher than that of the control group (P<0. 05). There were no severe adverse reactions occurred between two groups during the treatment period. The recurrence rate of amblyopia in the treatment group was significantly lower than that in the control group (P<0. 05) after 1a follow-up.· CONCLUSION: Visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia in preschool children can significantly improve patients' visual acuity, reduce the average annual diopter changes, improve the therapeutic effect, reduce the recurrence rate of amblyopia.

Objective To analyze host adaptability and drug resistance variation of highly pathogenic avian influenza (HPAI) A(H5N6) viruses obtained from outbreaks in Suzhou City. Methods Real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay was used for influenza virus detection in pharynx or anus swabs of dead birds and suspected cases. Full genome of H5N6 positive samples were sequenced by using Sanger method. Hemagglutinin (HA) and neuramidinase (NA) phylogenetic trees were constructed by MEGA software. Results A child was laboratory confirmed to have A(H5N6) viruses infection in November 2018. A outbreak of avian flu in poultry in W district during the Spring Festival was laboratory confirmed as a A(H5N6) epidemic situation. Avian-origin H5N6 viruses possessed D198N and Q226H resistance mutations. The homology of HA and NA genes of Suzhou strains were 98.01%-100.0% and 98.16%-100.0%, respectively. These H5N6 strains belonged to 2.3.4.4 H5 clad. Multiple basic amino acid at the cleavage site of HA implied the highly pathogenic characteristics of Suzhou H5N6 strains. Conclusion H5N6 is a kind of malti-sourse reassortment virus, which is still evolving. Multiple loci of Suzhou H5N6 strains of this epidemic were identified to have drifted. The prevalence of drug-resistant mutations should be closely monitored in order to timely take effective measures to prevent serious damage to public health and poultry production industries.

Objective: To elucidate the possible mechanism responsible for the improved protection of terminal warm blood cardioplegia (TWBC) after hypothermic cardiopulmonary bypass (CPB) through analysis of tubulin (TB) components changes in myocardial cells exposed to TWBC. Methods: Stable animal models of CPB were established in cats, which were then randomly divided into 2 groups. Group Ⅰ was subjected to intermittent cold blood cardioplegia (ICBC) whereas group Ⅱ to ICBC followed by TWBC before uncross-clamping. Left ventricular performance was then monitored and evaluated by LVSP, LVEDP, ±dp/dtmax and t-dp/dtmax in both groups and semi-quantitive analysis was conducted with Western blot method as to the content and constitution of TB in myocardial cells at 15 min, 120 min after aortic crossclamping (ACC) and 5 min,15 min, 60 min,120 min after reperfusion. Results: Within 120 min after reperfusion, systolic and diastolic functions decreased significantly in group Ⅰ as compared with group Ⅱ(P<0.05). At 115 min after ACC and 15 min after reperfusion, the content of free and polymerized TB in both groups had no difference (P>0.05). At 120 min after ACC and 5 minutes after reperfusion, there was a significant difference between groupⅠ andⅡ (P<0.01). Conclusion: TWBC accelerates the repolymerization of myocardial TB during hypothermic CPB, which may mediate the improved cardiac performance in the early stage of myocardial reperfusion.

Objective To investigate clinical and epidemiological features of pneumocystis jirovecii pneumonia (PJP) in kidney transplant recipients. Methods Clinical data of 68 kidney transplant recipients admitted from July, 2021 to December, 2021 were collected. All patients were divided into the PJP group (n=11), common pulmonary infection group (n=24) and non-pneumonia group (n=33) according to the status of pulmonary infection. The incidence and treatment of PJP after kidney transplantation were analyzed. Basic characteristics and laboratory parameters of the recipients were compared among all groups. The genotyping and transmission map of PJP patients were evaluated. Results Among 64 kidney transplant recipients, 11 cases were definitely diagnosed with PJP. The most common clinical manifestations included elevated body temperature, and dry cough complicated with progressive dyspnea. Chest CT scan showed diffuse interstitial inflammation and ground glass-like lesions of bilateral lungs in all patients. After diagnosis, all patients were orally given with compound sulfamethoxazole for 3-4 weeks. Two patients received non-invasive ventilator-assisted ventilation due to severe lung infection and dyspnea, and the remaining patients were given with nasal cannula oxygenation. One patient experienced elevated serum creatinine level upon discharge, and developed renal allograft failure. The remaining 10 recipients with PJP obtained normal renal allograft function, and no recipient died. Compared with the non-pneumonia group, the rejection rate was higher, the length of hospital stay was longer, the lymphocyte count was less, the lymphocyte proportion was lower, the levels of C-reactive protein, serum creatinine and lactate dehydrogenase were higher, and the levels of serum albumin was lower and CD4⁺T cell count was less in the PJP group (all P < 0.05). Compared with common pulmonary infection group, the lymphocyte count was less, the lymphocyte proportion was lower, the CD4⁺T cell count was less and 1, 3-β-D- glucan (BDG) level was higher in the PJP group (all P < 0.05). No new genotype was detected in 10 of the 12 testing samples. It was considered that PJP mainly depended on two transmission chains and two independent transmission individuals. Conclusions Kidney transplant recipients are prone to pneumocystis jirovecii (PJ) infection due to impaired cellular immune function. The most common clinical manifestations consist of elevated body temperature and dry cough complicated with progressive dyspnea, accompanied by headache and fatigue in partial patients. Chest CT scan shows diffuse interstitial inflammation and ground glass-like lesion of bilateral lungs. PJ may be transmitted through respiratory tract. Small-scale PJP might occur in the follow-up outpatient of kidney transplant recipients. Preventive measures should be delivered in a timely manner.

Transmission electron microscopy(TEM)is considered as an important characterization tool for revealing morphology of materials and an indispensable strategy for studying the mechanisms of charge-discharge process in battery.TEM samples needs be less than 0.1 μm thick,which means electrodeposited materials must undergo pre-treatment processes such as focusing ion beam etching,ultra-thin slicing,or ultrasonic dispersion before they can be observed via TEM.However,such treatments cause structure changes,and what real formed in electrodes is hard to estimate.In this work,a self-interlacing film layer composed of carbon nanotubes(CNTs)was fabricated on a copper grid through blade coating.A novel TEM grid was produced by optimizing the interlacing film's thickness and covering area through adjusting the interlacing state of various concentrations of CNTs.Utilizing the novel TEM grid as the battery's positive electrode,electrode deposits were acquired and subjected to TEM analysis to generate high-definition microstructure images of the electrode surface.This process provided new insights into sample preparation for investigating the deposition/stripping mechanism in high-energy-density metal anodes.

OBJECTIVETo compare the functional parameters of the small airways and clinical characteristics between patients with typical asthma (TA) and cough-variant asthma (CVA).

METHODSForty-three newly diagnosed asthmatic patients were enrolled, including 15 with TA and positive bronchial provocation test [TA BPT(+)], 12 with TA and positive bronchial dilation test [TA BDT(+)] and 16 with CVA, and 27 healthy subjects served as the control group. All the subjects were required to complete data acquisition, asthma control test, asthma control test scale, fractional exhaled nitric oxide, airway resistance and pulmonary function tests, BPT or BDT.

RESULTSThe interval from onset to a definite diagnosis of TA BDT(+) was longer than that of TA BPT(+), while that of CVA was the shortest (P=0.022). The pulmonary functional parameters of TA BDT (+) was significantly lower than those of the other 3 groups (P<0.05). MMEF, MEF, MEF, and MEFin patients with TA BDT(+), TA BPT(+) and CVA were significantly lower than those in the control group (P<0.01). The resonant frequency, respiratory impedance, resistance at 5 Hz, resistance at 20 Hz, and reactance at 5 Hz were significant higher in patients with TA BDT (+) than in the control subjects, while these parameters showed no significant differences among TA BPT (+), CVA and control groups. The airway resistance in TA BPT(+), CVA, and control groups increased after BPT, and the patients with TA BPT(+) showed greater changes in airway resistance than those in CVA and control groups. In CVA patients, FeNO showed a strong positive correlation with respiratory impedance (r=0.523, P=0.038), resistance at 5 Hz (r=0.542, P=0.030), and resistance at 20 Hz (r=0.524, P=0.037), and the airway responsiveness showed a strong positive correlation with resistance at 20 Hz (Rho=-0.512, P=0.043).

CONCLUSIONCVA is the early stage of TA, and CVA, TA BPT(+), and TA BDT(+) may represent different stages of asthma. Uncontrolled, prolonged CVA may evolve into TA BPT (+), whose further progression can cause damages of the pulmonary function and small airway function and leads eventually to TA BDT (+).

OBJECTIVETo investigate the content of nerve growth factor (NGF) in penis of rats with diabetic erectile dysfunction (DED) and apply rhNGF to treat the DED rat model to study the pathogenesis mechanism of DED and treatment effects by NGF for DED rat model.

METHODSFifty adult male SD rats were randomly selected to make up diabetic models. After 8 weeks, the mRNA and protein levels of NGF in rat penis were detected. Then the rest of rats were divided into 5 groups: normal control group, non-treated diabetes group, NGF treatment group, insulin treatment group, NGF plus insulin treatment group. After 8 weeks of treatment, intracavernous pressure (ICP) was measured. The content of neuronal nitric oxide synthase (nNOS) in the erectile tissue was also value- ated by immunohistochemistry staining.

RESULTSCompared with those of normal control group, the mRNA content of NGF and protein in the penis of non-treated diabetic rats is increased significantly. Compared with non-treated diabetic group, ICP was much higher in the treated groups which received NGF or/and insulin therapy, and the changes in nNOS staining of those groups were the same.

CONCLUSIONThe injury of pelvic splanchnic nerves in advanced stage of diabetes may lead to DED, which may be relevant to the abnormal of NGF level or the NGF receptors. The increased extent of NGF in penis of rat with DED suggests that erectile nerves be severely damaged by diabete and the increase of producing NGF couldn't compensate the needs for reproduction of the nerve fibers. The ability of combination of NGF with its receptor may also be damaged. It is helpful to use exogenous NGF to lessen the partly neuropathy and improve the erectile dysfunction of diabetic rats. The abnormal of NGF may play an important role in the pathogenesis of diabetic ED and its treatment.

Animals ; Diabetes Mellitus, Experimental ; complications ; Erectile Dysfunction ; drug therapy ; etiology ; metabolism ; Male ; Nerve Growth Factor ; biosynthesis ; genetics ; therapeutic use ; Penis ; metabolism ; RNA, Messenger ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley