To obtain and analyze the sequence of the nucleocapsid gene from bovine coronavirus, and to produce the fusion protein of the N gene in E.coli in order to use this recombinant protein for the study of bovine coronavirus. The N gene of BCV-DQ strain was amplified by RT-PCR, in which the primers were designed on the basis of N gene sequence of BCV-Mebus strain. The PCR products of 1 347 bp in length were cloned and sequenced, and then inserted into the prokaryotic vector pET30a. The recombinant plasmids were then transformed into Escherichia coli BL21 and identified by SDS-PAGE and Western blot assay. ELISA assay was optimized of N protein as the coating antigen to detect the viruses in the clinical samples. In comparison with 6 BCV strains in GenBank, the sequence identity was proved to be more than 98.3%. Result in SDS-PAGE showed that the fusion protein had a molecular weight of 60 ku, and could be specifically recognized by mouse serum against BCV. The indirect ELISA was used to test 256 serum samples collected from Heilongjiang province and 65.23% samples were positive. On testing field samples, an overall agreement of 95.31% was generated between the the neutralization test of viruses (VN) and indirect ELISA. It is apparent that the N gene was highly conservative and is expressed in E. coli in high level,also the prokaryotic expression products of this gene show a fine reactiongenicity in immune responses. It was also suggested that the N protein may be a useful antigen for sero-diagnosis and epidemiological investigation of BCV.

BACKGROUND:Recombinant bovine basic fibroblast growth factor is a manifold effect cytokine which can promote angiogenesis, wound healing, tissue repair and bone regeneration. Recombinant bovine basic fibroblast growth factor with good histocompatibility is easy to operate and has been widely used in oral and maxil ary surgery.
OBJECTIVE:To evaluate the effect of recombinant bovine basic fibroblast growth factor against dry socket syndrome after tooth extraction.
METHODS:A total of 160 patients who had been extracted mandibular third molar were selected and randomly divided into two groups. In the experimental group, recombinant bovine basic fibroblast growth factor was put into the sockets after mandibular third molars were extracted, while in the control group, we let the wounds to be healed natural y without any materials. The incidence of dry socket syndrome was observed and compared between two groups at 3 days, 5 days and 1 week after tooth extraction.
RESULTS AND CONCLUSION:One patient had dry socket after operation in the experimental group, and the incidence was 1.25%. In the control group, 10 patients suffered from dry socket, and the incidence was 12.5%. There was a significant difference in the incidence of dry socket between the two groups (P<0.01). There was visible granulation tissue within the tooth socket after tooth extraction in the experimental group, and extraction sockets narrowed and were fil ed with granulation tissues, which was 1-2 days earlier than the control group. No al ergies, tissue hyperplasia and other local and systemic reactions occurred in patients receiving implantation of recombinant bovine basic fibroblast growth factor gel. These findings indicate that local implantation of recombinant bovine basic fibroblast growth factor gel after mandibular tooth extractions can speed up the healing of dental extraction wounds.

OBJECTIVE To discover diversity of X gene sequences of hepatitis B virus isolates in hepatitis B induced liver cirrhosis patients and HBV carriers.METHODS DNA fragment including X gene sequences of hepatitis B virus was amplified,sequencing PCR products was preformed.The PCR products of three liver cirrhosis patients and three HBV symptomless carriers were cloned into pGEMT Easy vectors.Positive clones with target sequences were selected out for sequencing.Sequence comparison was made to find the identity.RESULTS A comparison of T1762/A1764,G1719T,T1727G/A,G1730C and T1753C mutations in a core promoter between the liver cirrhosis patients and the HBV carriers showed that the HBV isolates from the former had higher frequencies of mutation than the latter.The X promoter region of the HBV isolates from the liver cirrhosis patients showed higher frequencies of mutation than the isolates from the HBV carriers.Additionally,the homology between clones of X gene from one individual with liver cirrhosis averages 91.3-99.7%,that of HBV carriers averages 96-100%.CONCLUSIONS The core and X promoter region of the HBV isolates from the liver cirrhosis patients show the higher frequencies of mutation than the isolates from the HBV carriers.There are HBV quasispecies which possess great variation in the liver cirrhosis patients.

Objective To evaluate the role of calcium channel in the mechanism of the generation and maintenance of bursting firing of substantia nigra pars compacta (SNc) dopaminergic neurons in rats.Methods Using the patch clamp technique,we observed the firing pattern switching features after adding 10 μmol/L N-methyl-D-aspartic acid (NMDA),compared the changes of whole-calcium current and L-type calcium current with or without NMDA,and analyzed the correlation between the generation of burst firing and L-type calcium channel activation.Results After NMDA treatment,the firing pattern of SNc dopaminergic neurons changed to burst firing,which was compromised by a charastistic high plateau potential and series of action potential on it.The current density of L-type calcium current increased significantly after adding NMDA,which,from (2.86 ±0.26) pA/pF (n =28),significantly increased to (3.75 ± 0.18) pA/pF (n =34 ; t =7.52,P =0.002 8).The high plateau potential was almost abolished with the application of verapamil,a specific antagonist of L-type calcium channel.Consiusion NMDA could induce the firing pattern changed to burst firing in SNc dopaminergic neurons,while L-type calcium channel contributes to the process of generation and maintenance of burst firing.

Objective To study the differences of UVB damage among yong,intermediate and aged human dermal fibroblasts (HDFs),the recovery effect of adipose-derived stem cell conditioned medium (ADSC-CM) on UVB damage and the relationship between the recovery effect and ADSC-CM concentration as well as its treating period.Methods HDFs were cultured at different senescent degrees in vitro to establish UVB damage model of HDFs,and then the HDFs were cultured in different concentration ADSC-CM and cellular proliferation was detected.Results The optimal treating period and concentration of ADSC-CM were 48h and 100％ respectively.As to HDFs in different senescent degrees,ADSC-CM was able to accelerate cellular proliferation in both UVB group and control group.Conclusions Independently or simultaneously,the intrinsic and extrinsic factors can cause ageing damages on HDFs.To some extent,ADSC-CM has recovery effect on the damages through secretomes from ADSC.

OBJECTIVETo evaluate the efficacy and safety of meropenem in Chinese patients, we conducted a study for the treatment of patients with lower respiratory tract infections, urinary tract infections and other infections.

METHODSA total of 182 hospitalized patients were enrolled in the study. 90 patients received 500 mg meropenem every 12 hours (or 1 g every 12 hours if necessary) and 92 patients received imipenem/cilastatin 500 mg/500 mg every 12 hours (or 1 g every 12 hours if necessary) by intravenous infusion. The duration of treatment was 7 - 14 days for both groups.

RESULTSSeventy of 90 cases receiving meropenem and 70 of 92 cases receiving imipenem/cilastatin were assessable for clinical efficacy. The overall efficacy rates were 90% for the meropenem group and 87% for the imipenem/cilastatin group, and the bacterial eradication rates were 86% in both groups. 93 (76%) of 123 strains isolated from patients produced beta-lactamases. Adverse drug reactions were evaluated in 72 cases in the meropenem group and 70 cases in the imipenem/cilastatin group. The adverse drug reaction rates were 9.7% and 8.6%, respectively. The results showed that there were no statistical differences between these two groups (P > 0.05).

CONCLUSIONMeropenem is effective and safe for the treatment of bacterial infections caused mainly by beta-lactamase-producing strains.

Adult ; Aged ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Cilastatin ; administration & dosage ; adverse effects ; therapeutic use ; Female ; Humans ; Imipenem ; administration & dosage ; adverse effects ; therapeutic use ; Male ; Middle Aged ; Respiratory Tract Infections ; drug therapy ; Thienamycins ; adverse effects ; therapeutic use ; Urinary Tract Infections ; drug therapy

Objective:To explore the application value of sevoflurane inhalation anesthesia in mitral valve replacement.Methods:A total of 94 patients who underwent mitral valve replacement in our hospital (October 2016-October 2018) were randomly divided into the control group ( n=47) and the observation group ( n=47). The control group received target-controlled infusion of propofol, and the observation group inhaled sevoflurane.The postoperative conditions [intensive care unit (ICU) stay time, extubation time of tracheal tube, spontaneous cardiac rebound], hemodynamic indexes [mean arterial pressure (MAP), heart rate (HR)], serum creatine phosphokinase isoenzyme (CK-MB), cardiac troponin I (cTnI), malondialdehyde (MDA) and superoxide dismutase (SOD) in the two groups were analyzed. The patients were followed up for one month. The incidence of major adverse cardiovascular events (MACE) was calculated. Results:⑴ Postoperative situation: the time of stay in ICU and extubation of tracheal tube in the observation group was shorter than that in the control group, and the rate of spontaneous cardiac rebound (93.62%) was higher than that in the control group (72.34%) ( P<0.05); ⑵ Hemodynamic index level: there was no statistically significant difference in MAP and HR levels between two groups before operation, before cardiopulmonary bypass, after cardiopulmonary bypass, and after operation ( P>0.05); ⑶ CK-MB and cTnI: the levels of serum CK-MB and cTnI in the two groups were higher at 2, 6, 24, and 48 h after aortic cross-clamp release than before anesthesia induction, but the indicators of the observation group were lower than those in the control group; ⑷ MDA and SOD: the serum SOD level in the two groups at 2, 6, 24, and 48 h after aortic cross-clamp release were lower than before anesthesia induction, and the MDA level in the two groups at 2, 6, 24, and 48 h after aortic cross-clamp release were higher than before anesthesia induction. The level of SOD in the observation group was higher than that in the control group, and the level of MDA was lower than that in the control group ( P<0.05); ⑸ MACE: the incidence of MACE in the observation group (12.77%) was lower than that of the control group (29.79%) ( P<0.05). Conclusions:During mitral valve replacement, sevoflurane inhalation anesthesia can maintain hemodynamic stability. The duration of ICU stay and tracheal tube extubation time is shorter, and the fluctuation of serum CK-MB, cTnI, MDA and SOD is small, and it can reduce the risk of MACE.

Objective:To examine whether multiple-course prophylactic intravesical instillation chemotherapy is superior to single-course chemotherapy in preventing bladder tumor recurrence after radical nephroureterectomy(RNU)for primary upper tract urothelial carcinoma(UTUC).Methods:A comprehensive literature search was performed using the PubMed, Wanfang Data and China National Knowledge Infrastructure and all publications before March 2021 about clinical trials comparing the effects of single-course and multiple-course prophylactic intravesical instillation chemotherapy after RNU for UTUC were retrieved.Analysis was performed using Stata/SE 12.0.Results:Seven trials included a total of 998 patients, with 473 participants receiving multiple-course and 525 receiving single-course intravesical instillation chemotherapy.Bladder cancer recurrence occurred in 69 out of 473 patients(14.6%)with multiple-course chemotherapy and in 123 out of 525 patients(23.4%)with single-course chemotherapy.The absolute risk reduction was 8.8% and the relative risk reduction was 37.6%.Compared with a single-course instillation, the pooled OR of bladder cancer recurrence was 0.55(95% CI: 0.40-0.76, P<0.001)for multiple-course instillations.No serious adverse events were reported. Conclusions:Compared with single-course instillation chemotherapy, multiple-course prophylactic intravesical instillation chemotherapy significantly decreases the risk of bladder cancer recurrence in primary UTUC patients treated with RNU, indicating multiple intravesical instillation chemotherapy is superior to single instillation chemotherapy.

Objective:To evaluate the efficacy and safety of robotic arm assisted laparoscopic hysterosacral fixation in patients with pelvic organ prolapse(POP), and its impact on lower urinary tract function.Methods:This study retrospectively analyzed the clinical data of POP patients who had undergone robotic arm assisted laparoscopic hysterosacral fixation at our center from June 2019 to October 2020, and conducted exploratory research.Results:A total of 6 patients were included in the study, with POP quantitative staging above stage Ⅲ.The ages ranged from 70 to 82 years.The number of births each patient had given ranged from 1 to 3, & all were via vaginal deliveries.There were no significant changes in urodynamic parameter scores in any patients before and after surgery, but half of the patients had detrusor overactivity before surgery, which all disappeared after surgery.In addition, synchronous X-ray images showed that the postoperative pelvic organs were closer to the normal anatomical position.At the same time, quantitative staging of POP had achieved clear improvement, and related scale scores also significantly improved.One patient complained of occasional lumbar and back discomfort with postoperative over-stretching during outpatient review, which improved after symptomatic treatment.Conclusions:robotic arm assisted Laparoscopic hysterosacral fixation is satisfactory in efficacy and safety for POP patients, with good postoperative restoration of the uterus to the anatomical position and has insignificant influence on the function of the lower urinary tract.It is worth further assessment for wide application.

Objective:To explore the efficacy and safety of robot assisted laparoscopic sacrocolpopexy in the treatment of pelvic organ prolapse.Methods:The data of 16 patients who underwent robot assisted laparoscopic sacrocolpopexy in Beijing Hospital from September 2019 to May 2022 were collected. The average age was (73.5±9.3) years, the preoperative course was 4-240 months, the body mass index was (24.2±1.7) kg/m 2, the number of births was (1.7±0.8), and the quantitative index of pelvic organ prolapse (POP-Q) was grade Ⅲ -Ⅳ. The maximum urine flow rate before operation was (9.6±3.4) ml/s, the maximum detrusor pressure during urination was 26 (20, 32) cmH 2O, the first sense urine volume of the bladder was (119.1±39.4) ml, the first sense urine urgency volume of the bladder was (253.6±75.7) ml, the maximum bladder pressure capacity was (406.0±79.8) ml, and the residual urine volume was 10 (10, 28) ml. The preoperative PFDI-20 score was 100 (70, 122) and the PFIQ-7 score was 107 (90, 160). During the robot assisted laparoscopic sacrocolpopexy, the right area of the sacral promontory was exposed, the anterior and posterior walls of the uterus were separated, and the 2 cm small hole was separated from the right broad ligament of the uterus. The mesh was cut into a "Y" shape and passed through the small hole. The anterior and posterior leaves of the Y-shaped mesh were sutured to fix the cervix, and the other end was fixed to the anterior longitudinal ligament of the sacrum. The operation time and intraoperative bleeding were observed. The effect of surgery was evaluated by preoperative and postoperative urodynamic imaging, POP-Q stage, PFDI-20 score and PFIQ -7 score. Results:All the 16 operations were successfully completed. No injury of urethra, bladder, rectum, important blood vessels and nerves occurred during the operation. The average operation time was (255.6±56.0) min, and the average amount of bleeding was (28.8±18.2) ml. There was no inhibitory contraction wave in 7 patients (44%) before operation, suggesting that there was detrusor overactivity. After operation, the detrusor overactivity disappeared or significantly decreased in 7 patients. The postoperative follow-up period was 3-36 months. During the follow-up period, one patient had recurrence, and the rest had no prolapse and urination problems. The POP-Q stage was reduced to grade 0-Ⅰ after the operation. The subjective satisfaction rate of patients was 94%. The PFDI-20 score [13(8, 24)] and PFIQ -7 score [11(6, 15)] after operation were significantly reduced ( P<0.001), the initial urgent volume of bladder (272.5±88.5) ml was significantly increased compared with that before operation ( P=0.038), and the maximum volume of bladder (427.2±79.2) ml was significantly increased compared with that before operation ( P=0.006). Image urodynamics showed that the patient basically recovered the pelvic floor anatomy and achieved functional reduction. Conclusions:Robot assisted laparoscopic sacrocolpopexy has good subjective and objective effects in POP, low recurrence rate and less complications. It needs a larger sample size study for confirming the improvement of bladder function.