[Correlation between sonographic appearance of experimental testicular ischemia and histological changes of the testis after reperfusion].

OBJECTIVE: To discuss the correlation between sonographic appearance of different degrees of acute unilateral testicular ischemia and histological changes of the testis after reperfusion. METHODS: Thirty-two male rabbits were equally divided into a control (sham operation) group and 3 ischemia groups. Unilateral testicular ischemia models were established under the color Doppler ultrasound (CDU) observation and allocated according to different degrees of ischemia to Group A (with homogeneous echoes and slightly decreased flow signals), B (with heterogeneous echoes and obviously decreased flow signals) and C (with radial or small-shredded low echoes and absence of flow signals). Then contrast-enhanced ultrasonography (CEUS) was performed, followed by reperfusion of the ischemic testes. A month later, the histological changes of the testes were observed and the correlation of the histological changes with the sonographic and CEUS manifestations was analyzed. RESULTS: Concerning the CEUS parameters such as the arrival time (AT), reperfusion rate, time-to-peak (TTP), half descent time (DT/2) of the ischemic testes, Groups A and B showed significant differences from the control group (P < 0.05). The peak-base difference (PBD) was significant in Group B (P < 0.05) but not in A (P > 0.05), and no enhancement was seen in Group C. As for Johnson's scores obtained 1 month later, Group A exhibited no significant difference (8.70 +/- 0.39) (P > 0.05), Group B showed significant difference (6.01 +/- 0.88) (P < 0.05), and Group C extremely significant difference (3.16 +/- 1.05) (P< 0.001) from the control group (9.10 +/- 0.11). CONCLUSION: CEUS is superior to CDU in evaluating the perfusion of testicular ischemia. Sonographic appearances of testicular ischemia are significantly correlated with histological changes of the testis after reperfusion. Ultrasonography helps to predict the spermatogenetic function of ischemic testes after reperfusion.

Ultrasonographic diagnosis of hepatic fungal infection.

BACKGROUND: Hepatic fungal infection is difficult to diagnose early because of the lack of related clinical knowledge. In this study, we analyzed the ultrasonic manifestations of hepatic fungal infection, in order to determine the diagnostic value of ultrasonography. METHODS: In 8 patients with hepatic fungal infection, the results of color Doppler flow images (CDFI) and contrast-enhanced ultrasonography (CEUS) were compared with those of ultrasound-guided biopsy. RESULTS: In the 8 patients with pathologically proven hepatic fungal infection, 5 showed multiple hypoechoic lesions with unequal sizes in the liver, commonly less than 2 cm in diameter. Some of the hypoechoic lesions showed a hyperechoic strip in the periphery, while a minority showed a "bull's eye" pattern, which is considered a typical manifestation of metastatic hepatic carcinoma. Solitary lesions in the right lobe of the liver were found in the other 3 patients. No blood flow signals were found in any lesions with CDFI. In the lesions a "filling defect" pattern was shown by CEUS, indicating no enhancement in the arterial phase, the portal vein phase and the delayed phase, except for 3 patients, in whom the lesions were slightly enhanced in the periphery during the early arterial phase. CONCLUSIONS: Hepatic fungal infection is characterized by an ultrasonic manifestation of multiple hypoechoic lesions with inequal size. A small number of lesions may show a characteristic "bull's eye" appearance. Moreover, a "filling defect" pattern on CEUS is helpful to make a correct diagnosis. In addition, ultrasound-guided biopsy is an important method for accurate diagnosis of hepatic fungal infection.

Color Doppler velocity profile and contrast-enhanced ultrasonography in assessment of liver cirrhosis.

BACKGROUND: This study was designed to probe the clinical value in assessing the degree of liver cirrhosis by using the arrival time of contrast agent in the right portal vein in contrast-enhanced ultrasonography, as well as the velocity and flow volume in the right portal vein using the color Doppler velocity profile technique. METHODS: Twenty-eight patients with HBV post-hepatic cirrhosis were grouped into compensated (13 patients) and decompensated cirrhosis (15); 30 patients without hepatic cirrhosis served as controls. Written informed consent was obtained from each patient. All the patients with hepatic cirrhosis were pathologically confirmed by percutaneous biopsy. SonoVue was injected to detect the arrival time in the right portal vein. The velocity and flow volume in the right portal vein were measured. The value of each parameter was compared for correlation analysis. RESULTS: The arrival time in the right portal vein in the cirrhosis group was much longer than that in the control group (24.92+/-1.34 vs. 20.81+/-0.55 sec, respectively, P<0.01). The mean velocity, maximal velocity and flow volume in the cirrhosis group were much lower than those in the control group (10.64+/-0.84 vs. 14.78+/-0.71 cm/sec, 13.68+/-1.02 vs. 17.30+/-0.68 cm/sec and 358.72+/-23.63 vs. 438.61+/-16.86 ml/min, respectively, P<0.01). With the development of cirrhosis, the arrival time in the right portal vein was longer (P<0.05), and the velocity and flow volume was lower (P<0.01). There was a negative correlation between arrival time and mean velocity, maximal velocity and flow volume in the right portal vein in the cirrhosis group (r=-0.547, P<0.01; r=-0.508, P<0.05; r=-0.471, P<0.05, respectively). CONCLUSIONS: With the development of liver cirrhosis, the arrival time of contrast agent in the right portal vein is gradually prolonged, whereas the velocity and flow volume in this vein decreases markedly, and there is a negative correlation between the results of the two methods.

[Relationship between ultrasonographic grading of varicocele and changes of seminal parameters after varicocelectomy].

OBJECTIVE: To discuss the correlation of ultrasonographic grading of varicocele (VC) with the changes of seminal parameters after varicocelectomy. METHODS: A total of 129 cases of VC were diagnosed by scrotal color Doppler ultrasonography (CDU) and graded into 3 groups: VC I, VC II and VC III, according to three different grading indexes: the regurgitant volume of pampiniform plexus veins (PPV), the maximum diameter of PPV and the combination of the regurgitant volume and the maximum diameter of PPV. All the patients underwent high ligation of the spermatic vein and semen analysis was performed before and after varicocelectomy according to the WHO guidelines. RESULTS: With the maximum diameter of PPV as the grading index, there were statistically significant differences in the seminal parameters of VC III (P < 0.05) but not in those of VC I and VC II (P > 0.05) before and after varicocelectomy, while based on the regurgitant volume of PPV or the combination of the regurgitant volume and the maximum diameter of PPV, the differences were insignificant in VC I (P > 0.05), significant in VC II (P < 0.05) and extremely significant in VC III (P < 0.01). No significant differences were found in the rate of sperm abnormality among the 3 groups after varicocelectomy (P > 0.05). CONCLUSION: Different grading indexes influence the changes of seminal parameters after varicocelectomy. The regurgitant volume of PPV and the combination of the regurgitant volume and the maximum diameter of PPV are more reasonable and reliable as the grading indexes of VC. Different grades of VC respond differently to varicocelectomy. Whether VC I necessitates varicocelectomy needs to be further investigated.

Effect of fatty liver background on contrast-enhanced ultrasonographic appearance of focal nodular hyperplasia.

BACKGROUND: Focal nodular hyperplasia (FNH) is increasingly diagnosed as a result of the advances in imaging studies such as contrast-enhanced ultrasonography (CEUS), enhanced computed tomography and magnetic resonance imaging. However, FNH with atypical features can be difficult to differentiate from other benign and malignant tumors. The aim of this study was to investigate the influence of fatty liver background on the CEUS characteristics of FNH. METHODS: Twenty-six patients with FNH were divided into two groups: group A included 14 patients with fatty liver and group B included 12 patients with normal liver background. Conventional two-dimensional ultrasonography and color Doppler flow imaging (CDFI) were conducted and followed by real-time dual-frame CEUS. RESULTS: On two-dimensional ultrasonography, hypo-echoic nodules were present in most of the patients in group A (12/14) and hyperechoic nodules in most of those in group B (7/12). The difference in the nodule echotextures between the two groups was statistically significant (P<0.05). Nodules with centrifugal blood flow signals on CDFI were found in 6 of the 14 patients in group A and 5 of the 12 in group B (P>0.05). On CEUS, nodules with a central spoked-wheel-like enhancement pattern in the early arterial phase were observed in 8 patients in group A and those with an eccentric enhancement pattern in the remaining 6 patients. In this group, 3 patients had hypoechoic nodules in the delayed phase. Eleven of the 14 patients in this group were diagnosed accurately with CEUS. In group B, nodules with a rapid central spoked-wheel-like enhancement pattern in the early arterial phase were found in 8 patients by CEUS and those with rapid an eccentric enhancement pattern in 4. The nodules were found to be continuously enhanced in the delayed phase. All of the patients in group B were accurately diagnosed with CEUS. CONCLUSIONS: A FNH nodule on a background of fatty liver may present a hypoechoic pattern on two-dimensional ultrasonography and a hypoechoic wash-out pattern in the delayed phase on CEUS. At this time, punch biopsy is needed for the diagnosis or differential diagnosis of FNH.

Ultrasound-guided fine needle biopsy of intrahepatic nodules and low elevation of AFP in early diagnosis of hepatocellular carcinoma.

BACKGROUND: With the progress in early clinical treatment of hepatocellular carcinoma (HCC), early detection and diagnosis of HCC have been increasingly pressing. Combined alpha-fetoprotein(AFP) determination and ultrasonography has become the main method for the detection of small HCC; but the relationship between low elevation of AFP and pathologic findings of small HCC has not been well defined. The aim of this study was to assess the value of ultrasound-guided fine needle biopsy of intrahepatic nodules and low elevation of serum AFP in the early diagnosis of HCC. METHODS: Fifty-nine patients with serum AFP exceeding 20 ng/ml and intrahepatic nodules varying from 0.8 cm to 5.0 cm in diameter who had been detected by ultrasonography underwent ultrasound-guided percutaneous fine needle biopsy, and cytological staining and histological sectioning were performed at the same time. RESULTS: Among the 59 patients, 11 patients (18.6%) showed AFP level above 400 ng/ml, 5 (8.5%) from 200 ng/ml to 400 ng/ml, 18 (30.5%) from 50 ng/ml to 200 ng/ml and 25 (42.4%) from 20 ng/ml to 50 ng/ml. Follow-up demonstrated that 53 patients (89.8%) had a progressive increase of AFP level. In 58 patients (98.3%) cancer cells were found by cytological staining and/or histological sectioning. CONCLUSIONS: In those patients with slightly increased or continuously positive AFP, hepatic carcinoma should be highly suspected when AFP increases gradually and intrahepatic nodules are detected by ultrasonography in follow-up. Once intrahepatic carcinoma nodules are suspected, ultrasound-guided fine needle biopsy should be performed as early as possible for early diagnosis and treatment.

Ultrasound-guided fine needle aspiration biopsy in differential diagnosis of portal vein tumor thrombosis.

BACKGROUND: Portal vein tumor thrombosis (PVTT) is a serious complication and a major metastatic way of hepatocellular carcinoma(HCC). But portal vein benign thrombosis(PVBT) always appears in patients with hepatocirrhosis, and PVTT should be differentiated from PVBT. The aim of this study was to probe the value of ultrasound-guided fine needle aspiration biopsy in differential diagnosis of PVTT. METHODS: Twenty-two HCC patients with portal vein thrombosis and 8 hepatocirrhosis patients with portal vein thrombosis were studied by ultrasound-guided fine needle aspiration biopsy. Twelve portal vein thrombosis filling portal vein embranchment of the 30 portal vein thrombosis patients were examined by 18G automatic biopsy. The positive rates of aspiration biopsy cytology and histology were calculated and compared with those of automatic biopsy. RESULTS: The positive rates of fine needle aspiration biopsy cytology and histology were 93.3% (28/30) and 90.0% (27/30), respectively. They were not different markedly from that of automatic biopsy 91.7% (11/12). In aspiration biopsy of 22 HCC patients with PVTT, HCC cellular was found in 19 portal vein thrombosis patients (86.4%) by cytology examination and in 18 portal vein thrombosis patients (81.8%) by histology examination. In total, 20 tumor thrombi were detected. The other two were diagnosed as benign thrombosis. No HCC cell and/or tissue was observed in 8 patients with hepatocirrhosis associated with portal vein thrombosis. CONCLUSIONS: Ultrasound-guided fine needle biopsy in detecting PVTT shows a high positive rate and is of diagnostic value. The positive rate is not apparently different from that of automatic biopsy. Hence the case that fails to be diagnosed by color Doppler flow imaging(CDFI) and pulsed Doppler can be detected early by ultrasound-guided fine needle aspiration biopsy.

Experimental study on ultrasound-guided intratumoral injection of "Star-99" in treatment of hepatocellular carcinoma of nude mice.

AIM: To investigate the anti-cancer effect and the immunological mechanism of ultrasound-guided intratumoral injection of Chinese medicine "Star-99" in hepatocellular carcinoma (HCC) of nude mice. METHODS: Twenty-eight human hepatocellular carcinoma SMMC-7721 transplanted nude mice, 14 of hypodermically implanted and 14 of orthotopic liver transplanted, were randomly divided into three groups of which 14 mice with Star-99, and 7 with ethanol and saline respectively. Ten days after the transplantation the medicines were injected into the tumors of all the nude mice once every 5 days. After 4 injections the nude mice were killed. The diameters of three dimension of the tumors were measured by high frequency ultrasound before and after the treatment and the tumor growth indexes(*) (TGI) were calculated. Radioimmunoassay was used to detect the serum levels of interleukin-2 (IL-2) and tumor necrosis factor (TNF)-alpha. The tumor tissues were sent for flow cytometry (FCM) DNA analysis. Apoptotic cells were visualized by TUNEL assay. All the experiments were carried out by double blind method. RESULTS: The TGI of Star-99 group (0.076+/-0.024) was markedly lower than that of the saline group (4.654+/-1.283)(P<0.01). It also seemed to be lower than that of the ethanol group (0.082+/-0.028), but not significantly different (P>0.05). Serum levels of IL-2 and TNF-alpha were markedly higher than those of ethanol group and saline groups (P<0.05). The mean apoptotic index (AI: percentage of TUNEL signal positive cells) in Star-99 group (48.98+/-5.09 %) was significantly higher than that of the ethanol group (11.95+/-2.24 %) and the saline group (10.48+/-3.85 %) (P<0.01). FCM DNA analysis showed that the appearance rate of the apoptosis peak in Srar-99 group was 92.9 %, markedly higher than that of the ethanol group (14.3 %) and the saline group (0.0 %) (P<0.01). Correlation (r=0.499, P<0.05) was found between AI and serum level of TNF-alpha. CONCLUSION: Star-99 has an effect on the elevation of the serum levels of IL-2 and TNF-alpha. It indicates that Star-99 has the function of enhancing the cellular immunity and inducing cancer cell apoptosis. The correlation between AI and serum level of TNF-alpha indicates that the elevation of the serum of TNF-alpha induced by Star-99 may be an important factor in the promotion of the hepatic cancer cell apoptosis. Star-99 has strong effects on the inhibition and destruction of cancer cells. Its curative effect is as good as ethanol. Its major mechanisms can be as follows: (1) it increases the serum levels of IL-2 and TNF-alpha and triggers cellular immunity. (2) It can induce cancer cells apoptosis, the effective mechanism of the Star-99 is different from that of the ethanol. The mechanisms of triggering the immunologic function of the organism and inducing cell apoptosis are, of particular significance. This study will provide a new pathway of drug administration and an experimental basis for the treatment of HCC with Chinese herbal, and the study of Star-99 in the treatment of tumor is of profound significance with good prospects.

Biological characteristics of HCC by ultrasound-guided aspiration biopsy and its clinical application.

AIM: To probe the pathological biological characteristics of hepatocellular carcinoma (HCC) by the ultrasound-guided aspiration biopsy and assess the clinical application value of this method. METHODS: The biopsy and DNA analysis by flow cytometry (FCM) were taken in 46 cases with HCC nodules, including 26 cases and 20 cases with nodules < or =3 cm and >3 cm in diameters respectively, and 12 cases with intrahepatic benign hyperplastic nodules. They were taken in 22 cases of 46 cases with HCC before and after the therapy. Fine-needles and automatic histological incised biopsy needles were used. The fresh biopsy tissue was produced into the single cell suspension, which was sent for DNA detection and ratio analysis of cell period. The ratio of each DNA period of cell proliferation of each group was calculated and compared with each other. The DNA aneuploid (AN) and apoptosis cell peak were observed and their percentages were calculated. RESULTS: The ratios of S and G(2)/M periods of DNA, which reflect cell hyperproliferation, in the group with HCC tumors >3 cm in diameter were markedly higher than those of the group with HCC nodules < or =3 cm in diameter and the group with the benign hyperplastic nodules (P<0.01 except A:B of S period, P<0.05). The ratios of the middle group were also apparently higher than those of the latter group (P<0.01). The ratio of DNA AN of 46 cases with HCC nodules was 34.8 % (16/46). None of the cases with the intrahepatic hyperplastic nodules appeared AN. The DNA AN appeared more apparently with the growth of the tumors. The AN ratio of the group with tumors >3 cm in diameter was 55 % (11/20), markedly higher than that of the group with tumors < or =3 cm in diameter which was 19.2 % (5/26) (P<0.01). The FCM DNA analysis of 22 specimens of hepatic carcinoma tissue before therapy showed that the aneuploid peaks appeared in 5 cases (22.7 %). The ratio of G(1) period rose after therapy while the S period and G(2)/M ratios fell (P<0.01). The aneuploid peak disappeared in the 5 cases after the therapy, while the apoptosis peaks in 12 cases (54.5 %) appeared. CONCLUSION: Addition to supply the information of the pathological morphology of the tumor, the ultrasound-guided fine-needle aspiration tissue could be sent for FCM DNA analysis to comprehend its pathological biological characteristics. This can not only provide the clinic the reliable information about the occurrence, development, diagnosis, curative effect and prognosis of tumors but also supply biological information for clinic to choose therapeutic schemes.

Experimental and clinical assessment of percutaneous hepatic quantified ethanol injection in treatment of hepatic carcinoma.

AIM: To detect the relationship between absolute ethanol injection quantity, the interval and formation of fibreboard, the curative effect in treatment of hepatocarcinoma and to evaluate the clinical application of percutaneous hepatic quantified ethanol injection (PHQEI) in treatment of hepatic carcinoma (HCC). METHODS: (1) Experimental study: Twenty-four human hepatic carcinoma SMMC-7721 xenografted nude mice were randomly divided into three groups: group A injected with quantified ethanol at short intervals (QESI), group B with quantified ethanol at long intervals (QELI) and group C with a small quantity of ethanol at long intervals (SQLI). The tumor tissues were sent for patho-histology and electron microscopic examinations. The diameters of tumors were measured with high frequency ultrasound before and after therapies and tumor growth index (TGI) was calculated. (2) Clinical study: Tumors of 122 cases of pathologically proved HCC were injected with quantified ethanol guided by ultrasound every 3-5 d 4-10 times per period of treatment. The quantity of ethanol was calculated according to the regressive equations where Y = 2.885X when the mass was < or =5 cm in diameter and Y = 1.805X when the mass was >5 cm in diameter (X is the maximal diameter of the mass with the unit cm, Y is the ethanol quantity with the unit mL). The survival rates of 1, 2, 3 and 4 years and recurrent rates in situ as well as dystopia in the liver were calculated. RESULTS: (1) Experimental study: TGI of QESI group (0.072+/-0.018) and QELI group (0.094+/-0.028) was apparently lower than that of SQLI group (1.982+/-0.482) (P<0.01). TGI of QESI group seemed to be lower than that of QELI group, but it was not markedly different (P>0.05) between two groups. Severe degeneration and necrosis could be seen in QESI group by patho-histology examination. Coagulative necrosis could be seen in most tumors of QESI group and there were no residual cancer cells under electronic microscope, while the residual cancer and inflammatory cells and fibre tissues could be seen around the tumors of QELI group. Infiltration of inflammatory cells could be seen and fibre tissues were formed. (2) Clinical study: B mode ultrasound showed that 62.5% of tumors shrank after PHQEI. The survival rates of 1, 2, 3 and 4 years of the group with tumors < or =3 cm in diameter were higher than those of the group with tumors >3 cm in diameter. The recurrent rates of tumors in situ of the former group were apparently lower than those of the latter group. The recurrent rates of tumors in dystopia in the liver of the former group were markedly lower than those of the latter group. The 122 cases underwent a total of 1221 PEI. There were no complications such as hemorrhage and severe heart, liver and kidney functional injuries except for 1 case of melena and 4 cases of jaundice who recovered after 1-2 wk under common therapies. CONCLUSION: The experimental study shows quantified ethanol at intervals of 3-5 d could improve the curative effect of hepatocarcinoma. The clinical study shows PHQEI is an effective therapeutic method for HCC with few side-effects, and a low-cost. The treatment efficacy is more remarkable for tumors < or =3 cm in diameter.

Intraoperative ultrasonography in hepatobiliary surgery.

OBJECTIVE: To assess the value of intraoperative ultrasonography in hepatobiliary surgery. METHODS: Aloka-650 and Aloka-500 with a 3.5 MHz probe were used in preoperative ultrasonography and those with 5 MHz or 7.5 MHz probes in intraoperative ultrasonography. All patients with carcinoma were confirmed pathologically. RESULTS: In 44 patients with liver cancer (76 tumors), the diagnostic rate of intraoperative ultrasonography was 97% (74/76), which was markedly higher than 84% (64/76) of preoperative ultrasonography (P<0.05). The diagnostic rate (92%; 22/24) of intraoperative ultrasonography for tumors 0.05). Yet the diagnostic rate (100%; 12/12) of intraoperative ultrasonography for gallbladder carcinoma was markedly higher than that (66.7%; 8/12) of pre-operative ultrasonography (P<0.05). The diagnostic rates of intraoperative ultrasonography for choledocholith and biliary tract cancer were all 100%, markedly higher than those (83%; 20/24 and 75%; 12/16) of preoperative ultrasonography (P<0.05). CONCLUSIONS: Intraoperative ultrasonography can raise the detective rate of the liver occupying lesions comparing with that of preoperative untrasonography. It is of valuable in selecting operative scheme and locating small lesions.

Percutaneous intratumoral injection of traditional Chinese herbal compound medicine Star-99 in treatment of hepatocellular carcinoma of mice.

BACKGROUND: Unresectable hepatocellular carcinoma (HCC) lacks effective therapy and entails very poor progress. In 1991, we found that Chinese herbal compound Star-99 has potentially effect on HCC. The purpose of this study was to probe the anti-cancer effect and the mechanism of focal injection of Chinese herbal compound Star-99 into HCC of mice. METHODS: In 32 nude mice transplanted with human hepatocellular carcinoma SMMC-7721, 16 received hypodermic implant and the other 16 orthotopic liver transplant. They were randomly divided into three groups: Star-99 group (Chinese herbal compound, 16 mice), alcohol group (8) and saline group (8), respectively. Intratumoral injection of Star-99, alcohol and saline was carried out 10 days after transplantation of HCC. Twenty days after the first injection, the nude mice were killed after being injected every 5 days with a total of 4 injections in each mouse. Tumor tissues were examined pathologically or via an electron microscope and flow cytometrical (FCM) DNA analysis. The three diameters of the tumor were measured with high-frequency ultrasound before and after injection, and the growth index was calculated with the following formula: volume of tumor (after treatment-before treatment)/volume of tumor (before treatment). Double-blind method was applied in the experiment. RESULTS: The growth index of the Star-99 group (0.068+/-0.022) and the alcohol group (0.079+/-0.024) was markedly lower than that of the saline group (4.345+/-1.453, P<0.01), but there was no significant difference between the Star-99 and alcohol groups. Coagulation (8/8) was the major pathological change in the alcohol group. In the Star-99 group, however, the phenomenon of lymphocytes attacking cancer cells could even be seen under the electron microscope. The typical apoptosis cells and apoptosis bodies as well as the collagen fibrae lined in mass could also be seen in the group (14/16). FCM DNA analysis showed that the rate of apoptosis in the Star-99 group (93.8%) was significantly higher than that in the alcohol (12.5%) and saline groups (12.5%) (P<0.01). CONCLUSIONS: This study shows that Star-99 markedly inhibits and destructs hepatocellular cancer cells. Star-99 is effective to directly destroy the membrane, cytoplasm and nuclei of tumor cells, causing their crumbling, activate the immune function and inflammatory reaction of nude mice, and induce the apoptosis of cancer cells. The effect of Star99 is significantly different from that of alcohol that mainly causes coagulation of cancer cells. Star-99 is feasible in the treatment of HCC.