Nocardia is a Gram-positive pathogen responsible for causing dairy mastitis,which leads to purulent granulomatous lesions in mammary tissue and can significantly impact the dairy indus-try,resulting in substantial economic losses.To develop a rapid and accurate diagnostic method for detecting Nocardia of bovine origin,a conserved sequence of the 16S rRNA gene from Nocardia was selected from the NCBI database.Based on this sequence,a pair of primers and a TaqMan fluo-rescent quantitative probe were designed.The validation of the TaqMan fluorescence quantitative PCR(qPCR)method found in this study showed that the Ctvalue had a good linear relationship with recombinant plasmid concentrations ranging from 1×1010 to 1×102 copies/μL,with a regres-sion equation of y=-3.536x+43.78,a correlation coefficient(R2)of 0.997 5,a slope of-3.536,and an amplification efficiency(E)of 91％(where 90％＜E＜110％).The specificity was strong,with no cross-reactions with other pathogens.The standard curve had a high sensitivity with a low-er detection limit of 1 × 102 copies/μL,it was 100-fold higher than conventional PCR.The repeatability of the standard curve was also good.Both intra-and inter-group coefficients of varia-tion were below 2％.Using this method,234 milk samples and 80 environmental samples were tested using this method,respectively,with a positive detection rate of 27.07％,whereas conven-tional PCR had a positive detection rate of 19.43％,indicating that this method was more sensitive compared to conventional PCR.The fluorescent quantitative PCR detection method established in this study provides an effective means for the clinical detection of Nocardia in dairy cows.

Nocardia is a Gram-positive pathogen responsible for causing dairy mastitis,which leads to purulent granulomatous lesions in mammary tissue and can significantly impact the dairy indus-try,resulting in substantial economic losses.To develop a rapid and accurate diagnostic method for detecting Nocardia of bovine origin,a conserved sequence of the 16S rRNA gene from Nocardia was selected from the NCBI database.Based on this sequence,a pair of primers and a TaqMan fluo-rescent quantitative probe were designed.The validation of the TaqMan fluorescence quantitative PCR(qPCR)method found in this study showed that the Ctvalue had a good linear relationship with recombinant plasmid concentrations ranging from 1×1010 to 1×102 copies/μL,with a regres-sion equation of y=-3.536x+43.78,a correlation coefficient(R2)of 0.997 5,a slope of-3.536,and an amplification efficiency(E)of 91％(where 90％＜E＜110％).The specificity was strong,with no cross-reactions with other pathogens.The standard curve had a high sensitivity with a low-er detection limit of 1 × 102 copies/μL,it was 100-fold higher than conventional PCR.The repeatability of the standard curve was also good.Both intra-and inter-group coefficients of varia-tion were below 2％.Using this method,234 milk samples and 80 environmental samples were tested using this method,respectively,with a positive detection rate of 27.07％,whereas conven-tional PCR had a positive detection rate of 19.43％,indicating that this method was more sensitive compared to conventional PCR.The fluorescent quantitative PCR detection method established in this study provides an effective means for the clinical detection of Nocardia in dairy cows.

Objective:To provide an effective protection method for reducing the ankle injury during airborne force landing by investigating the protective effect of a protective ankle brace (PAB).Methods:Airborne soldiers were selected as subjects and they were asked to jump from 1.5 m and 2 m high training platform with and without PAB protection. They maintained a standard bending and knee-bending posture as they touched the buffer surface of the training ground. Each subject completed 4 simulated landing trials: from 1.5 m with PAB (1.5 m PAB group), from 2 m with PAB (2 m PAB group), from 1.5 m without PAB (1.5 m control group), and from 2 m without PAB (2 m control group). The muscle electrical signals of bilateral tibialis anterior, peroneal longus and brevis and gastrocnemius were recorded by surface electromyography, and the percentage of maximal voluntary electrical activation (MVE%) was calculated and analyzed.Results:Sixty-one subjects were included. At the same height with different ankle protection status, the MVE% values of bilateral tibialis anterior and gastrocnemius in the 1.5 m PAB group were lower than those in the 1.5 m control group, and the differences were significant ( t=2.45, 2.74, 2.35, P=0.017, 0.008, 0.022). The MVE% values of bilateral peroneus longus and brevis in the 1.5 m PAB group were higher than those in the 1.5 m control group, and the differences were significant ( t=5.28, 4.56, both P<0.001). The MVE% values of left tibialis anterior and left gastrocnemius in the 2 m PAB group were lower than those in the 2 m control group, and the differences were significant ( t=2.21, 0.53, 4.35, P=0.031, 0.598, <0.001). The MVE% values of bilateral peroneus longus and brevis in the 2 m PAB group were higher than those in the 2 m control group, and the differences were significant ( t=4.92, 6.31, both P<0.001). With the ankle protection the MVE% values of bilateral tibialis anterior, gastrocnemius, peroneus longus and brevis in the 1.5 m PAB group were lower than those in the 2 m PAB group, and the differences were significant ( t=3.56-4.94, all P≤0.001). The MVE% values of bilateral tibialis anterior, gastrocnemius, peroneus longus and brevis in the 1.5 m control group were lower than those in the 2 m PAB group, and the differences were significant ( t=2.30-5.85, all P<0.05). Conclusions:PAB can reduce the ankle injury during airborne force landing and provide good protection.

Objective:To provide an effective protection method for reducing the ankle injury during airborne force landing by investigating the protective effect of a protective ankle brace (PAB).Methods:Airborne soldiers were selected as subjects and they were asked to jump from 1.5 m and 2 m high training platform with and without PAB protection. They maintained a standard bending and knee-bending posture as they touched the buffer surface of the training ground. Each subject completed 4 simulated landing trials: from 1.5 m with PAB (1.5 m PAB group), from 2 m with PAB (2 m PAB group), from 1.5 m without PAB (1.5 m control group), and from 2 m without PAB (2 m control group). The muscle electrical signals of bilateral tibialis anterior, peroneal longus and brevis and gastrocnemius were recorded by surface electromyography, and the percentage of maximal voluntary electrical activation (MVE%) was calculated and analyzed.Results:Sixty-one subjects were included. At the same height with different ankle protection status, the MVE% values of bilateral tibialis anterior and gastrocnemius in the 1.5 m PAB group were lower than those in the 1.5 m control group, and the differences were significant ( t=2.45, 2.74, 2.35, P=0.017, 0.008, 0.022). The MVE% values of bilateral peroneus longus and brevis in the 1.5 m PAB group were higher than those in the 1.5 m control group, and the differences were significant ( t=5.28, 4.56, both P<0.001). The MVE% values of left tibialis anterior and left gastrocnemius in the 2 m PAB group were lower than those in the 2 m control group, and the differences were significant ( t=2.21, 0.53, 4.35, P=0.031, 0.598, <0.001). The MVE% values of bilateral peroneus longus and brevis in the 2 m PAB group were higher than those in the 2 m control group, and the differences were significant ( t=4.92, 6.31, both P<0.001). With the ankle protection the MVE% values of bilateral tibialis anterior, gastrocnemius, peroneus longus and brevis in the 1.5 m PAB group were lower than those in the 2 m PAB group, and the differences were significant ( t=3.56-4.94, all P≤0.001). The MVE% values of bilateral tibialis anterior, gastrocnemius, peroneus longus and brevis in the 1.5 m control group were lower than those in the 2 m PAB group, and the differences were significant ( t=2.30-5.85, all P<0.05). Conclusions:PAB can reduce the ankle injury during airborne force landing and provide good protection.

Objective: To explore the effect of hepatitis C virus (HCV) on the expression of serine protease inhibitor Kazal1 (SPINK1) and its clinical implication. Methods: mRNA and protein expression of SPINK1 in Huh7.5.1 cells infected by HCV JFH-1 and the control cells were measured by RT-PCR and western blotting, SPINK1 levels in the cell supernatants and sera of HCV patients were measured by enzyme-linked immunosorbent assay (ELISA), the difference of SPINK1 levels between healthy controls and HCV patients was analyzed. Results: Expression of SPINK1 mRNA and protein was higher in Huh7.5.1 cells infected by HCV JFH-1 than in the control cells, serum SPINK1 levels was much higher in HCV patients than in healthy controls (P=0.016). Conclusions HCV can upregulate the expression of SPINK1.

Objective To evaluate the feasibility of keeping the consistency of bladder filing by a Bladderscan ( BS ) device during radiotherapy for rectal cancer. Methods The bladder volume was measured using a BS device and recorded. To verify the reliability of the BS measurement, the urinary output was determined by a measuring glass after complete urinary discharge. The bladder volume of 42 patients determined by the planning computed tomography ( CT) scans was used as the standard urinary volume. The bladder volume was measured using the BS device before radiotherapy every day. The chief complaint urinary volume was determined as the bladder volume when a patient felt a strong urge to urinate. The controlled urinary volume was determined as the urinary output intervened by the BS device to reach the standard urinary volume. Comparison was made by t test and data was assessed by Pearson correlation analysis. Results There was no significant difference in obtained urinary volume between the BS device and the planning CT scan ( P=0?84) . The urinary volume measured by the measuring glass was correlated with that measured by the BS device ( P=0?00 ) . The ability to hold urine in all patients was gradually weakened during treatment. The controlled urinary volume had a significantly smaller decrease after 1, 2, 3, 4, and 5 weeks of treatment than the chief complaint urinary volume ( 5% vs. 21%;6% vs. 20%;6% vs. 20%, 4% vs. 21%;11% vs. 26%;all P=0?00) . Conclusions Patients have a gradually decreased urinary volume during treatment. The intervention by the BS device gives a bladder volume close to that in the planning system, which results in a consistent bladder filling.

Objective To study the effect of flupentioxl melitracen and pinaverium bromide treatment on the changes of anorectal motility and rectal sensation in the patients of diarrhea-predominant irritable bowel syndrome (IBS-D) accompanying with depression and/or anxiety status. Methods Forty-four patients with IBS-D accompanying with depression and/or anxiety status were divided into group A (flupentioxl melitracen and pinaverium bromide) and group B (pinaverium bromide) by random digits table,and treated for 4 weeks. Twenty-five healthy subjects were included as control group. The anorectal motility and rectal sensation before and after taking medicines were compared. Results When abdominal pressure was increased, the net increased pressure of anal sphincter was (3.0 ± 1.2 ) kPa in group A and (2.9 ± 1.2)kPa in group B. They were lower than that in control group [(3.6 ± 1.6) kPa](P＜ 0.05). The rectal lowest volume of sensory threshold, the maximal volume of tolerance and maximal compliance were (55 ± 20) ml,( 145 ± 78 ) ml, ( 21.9 ± 12.9 ) ml/kPa in group A, ( 56 ± 38 ) ml, ( 150 ± 50 ) ml, (20.8 ± 11.2) ml/kPa in group B. They were lower than those in control group [(80 ± 38 ) ml, ( 190 ± 50 ) ml, (30.8 ± 15.2 ) ml/kPa](P ＜ 0.01 ). The rectal lowest volume of sensory threshold, the maximal volume of tolerance and maximal compliance were higher than those before taking medicines. Only the rectal lowest volume of sensory threshold in group B was higher than that before taking medicines. The rectal lowest volume of sensory threshold, the maximal volume of tolerance and maximal compliance in group A after taking medicines were higher than those in group B (P ＜ 0.05 or ＜ 0.01 ). Conclusions Higher sensitivity, lower tolerance,lower compliance of rectum and weakened anal automatic control function in IBS-D may be associated with diarrhea and frequent defecation. Treatment combining flupentioxl melitracen with pinaverium bromide may preferably improve the aperception functions of rectum in the patients of IBS-D accompanying with depression and/or anxiety status.

Objective To report our experience on video-assisted thoracoscopic surgery(VATS)and perioperative management for patients over 70 years old.Methods From May 1998 to May 2008,78 patients over 70 years old were treated by VATS.Operative risk was evaluated by measuring forced expiratory volume in first second(FEV1),diffuse capacity of carbon monoxide(DLCO),and left ventricular ejection fraction(LVEF),and artery blood gas analysis and exercise test before the operation.VATS(n=46)or video-assisted minithoracotmy(n=32)was carried out under general anesthesia with a double-lumen tube.Extubation was delayed in 11 patients after the procedures and mechanical ventilation was employed to smooth anesthesia recovery.Results Postoperative pathological examination showed primary lung cancer in 20 cases,benign pulmonary lesions in 26,pleural mesothelioma in 3,malignant pleural metastasis in 5,empyema in 5,benign mediastinal tumor in 15,hiatal hernia in 3,and esophageal leiomyoma in 1.Postoperative morbidity was 33.3%(26/78)in this series,none of the patients died during the operation;the perioperative mortality was 2.6%(2/78).Conclusions VATS is safe for elderly patients.Satisfying outcomes can be achieved through strict preoperative evaluation,proper operative method and active management for postoperative complications.